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Sample records for stored blood culture

  1. Storing Blood Cells

    NASA Technical Reports Server (NTRS)

    1976-01-01

    The National Cancer Institute worked with Goddard Space Flight Center to propose a solution to the blood-cell freezing problem. White blood cells and bone marrow are stored for future use by leukemia patients as a result of Goddard and Jet Propulsion Laboratory expertise in electronics and cryogenics. White blood cell and bone marrow bank established using freezing unit. Freezing unit monitors temperature of cells themselves. Thermocouple placed against polyethylene container relays temperature signals to an electronic system which controls small heaters located outside container. Heaters allow liquid nitrogen to circulate at constant temperature and maintain consistent freezing rate. Ability to freeze, store, and thaw white cells and bone marrow without damage is important in leukemia treatment.

  2. Blood culture

    MedlinePlus

    Culture - blood ... A blood sample is needed . The site where blood will be drawn is first cleaned with an antiseptic such ... organism from the skin getting into (contaminating) the blood sample and causing a false-positive result (see ...

  3. Blood Culture (For Parents)

    MedlinePlus

    ... Kids to Be Smart About Social Media Blood Culture KidsHealth > For Parents > Blood Culture Print A A ... adjust the treatment choice. Why Do a Blood Culture? During some illnesses, certain infection-causing bacteria and ...

  4. Blood Culture Test

    MedlinePlus

    ... Complete Blood Count , Urine Culture , Bacterial Wound Culture , Gram Stain , CSF Analysis , Fungal Tests , Susceptibility Testing At a ... Other related tests that may be performed include: Gram stain —a relatively quick test used to detect and ...

  5. Blood culture contaminants.

    PubMed

    Dawson, S

    2014-05-01

    Blood cultures are an essential diagnostic tool. However, contamination may impact on patients' care and lead to increased patient stay, additional tests, and inappropriate antibiotic use. The aim of this study was to review the literature for factors that influence the rate of blood culture contamination. A comprehensive literature search was performed using Medline and CINAHL on blood culture contamination. Hospitals/units should have in place a protocol for staff on how to take blood cultures, incorporating use of an aseptic technique. Studies have shown that several key factors in the process may lower contamination rates such as adherence to a protocol, sampling by peripheral venepuncture route rather than via an intravascular catheter, use of sterile gloves, cleaning tops of blood culture bottles with antiseptics and inoculating blood culture bottles before other blood tubes, samples being taken by a phlebotomy team, monitoring contamination rates, and providing individual feedback and retraining for those with contaminants. Although skin antisepsis is advocated there is still debate on which antiseptic is most effective, as there is no conclusive evidence, only that there is benefit from alcohol-containing preparations. In conclusion, hospitals should aim to minimize their blood culture contamination rates. They should monitor their rate regularly and aim for a rate of ≤3%.

  6. Blood donations, iron stores, and risk of Parkinson's disease.

    PubMed

    Logroscino, Giancarlo; Chen, Honglei; Wing, Al; Ascherio, Alberto

    2006-06-01

    Iron overload and systemic iron stores may be important in the pathogenesis of Parkinson's disease (PD). We therefore examined the association between blood donations, which reduce body iron stores, and risk of PD in the Health Professionals Follow-Up Study, a large cohort investigation of U.S. men. Our hypothesis was that blood donation reduces the risk of PD by lowering systemic iron stores. Although the number of blood donations was inversely related to the ferritin levels in a subsample of the study population, no association was found between the number of blood donations and risk of PD (P for trend = 0.6). Unexpectedly, the risk of PD was higher among men who reported recent multiple blood donations (P for trend = 0.05). The results of this study do not support the hypothesis that reduced systemic iron stores lower the risk of PD.

  7. Blood Culture (For Parents)

    MedlinePlus

    ... What Other Parents Are Reading Your Child's Development (Birth to 3 Years) Feeding Your 1- to 3-Month-Old Feeding ... blood culture is a test that looks for germs such as bacteria or fungi in the blood. A doctor might order this test when a child has symptoms of ...

  8. [Blood culture update].

    PubMed

    Muller-Serieys, C; Bergogne-Bérézin, E

    2002-01-12

    Blood culture is one of the most important bacteriological examinations with important clinical and therapeutic consequences. Blood cultures should be ordered in all patients with signs suggesting septicemia, endocarditis or severe infection (pneumococcal pneumonia, bacterial meningitis with bloodstream dissemination). Blood culture methods have evolved considerably over the last twenty years. After using manual methods for many years, read by non-standardized visual methods, the development of media with defined compositions and supplemented to allow growth of bacteria difficult to culture has been associated with the development of automatic blood culture devices. These devices have undergone rapid improvement. Semi-automatic devices (Bactec NR-660) were rapidly followed by completely automatic techniques, including four devices currently available: since 1989 Bio-Argos (Rio-Rad) and Bact/Alert (Organon-Teknika) and in 1993, Bactec 9240 (Becton-Dickinson) and Vital (BioMérieux). All these devices allow automatic detection of CO2 produced during bacterial growth. Automatic reading systems provide continuous output avoiding the need for invasive methods and thus the risk of contamination in addition to saving time. Potential application to achieve quantitative blood cultures for intensive care units is in the development stage. The reliability of these devices is well recognized and their contribution to severe bacterial infection is undeniable. There are certain limitations however related to material cost and the non-identification of the pathogen involved. Molecular biology techniques open new perspectives in this field. The evolution of techniques, definitions, and pathogenic approach to septicemia must be revisited as new infectious situations have been identified at the same time as new investigation tools resulting from considerable technological progress. New methods of blood culture have largely contributed to this progress.

  9. Iron stores, blood donation, and insulin sensitivity and secretion.

    PubMed

    Fernández-Real, José Manuel; López-Bermejo, Abel; Ricart, Wifredo

    2005-07-01

    Epidemiologists have observed that blood donation is associated with decreased risk of type 2 diabetes and cardiovascular disease. We investigated the relationship between iron stores and insulin sensitivity, after controlling for known confounding factors, and compared insulin sensitivity between blood donors and individuals who had never donated blood (nondonors). In 181 men, insulin sensitivity and insulin secretion were evaluated through frequently sampled intravenous glucose tolerance tests with minimal model analysis. Men who donated blood between 6 months and 5 years before inclusion (n = 21) were carefully matched with nondonors (n = 66) for age, body mass index, waist-to-hip ratio, and cardiovascular risk profile, including blood lipids, blood pressure, and smoking status. Frequent blood donors (2-10 donations) had increased insulin sensitivity [3.42 (1.03) vs 2.45 (1.2) x 10(-4) x min(-1) x mIU/L; P = 0.04], decreased insulin secretion [186 (82) vs 401.7 (254) mIU/L x min; P <0.0001], and significantly lower iron stores [serum ferritin, 101.5 (74) vs 162 (100) microg/L; P = 0.017] than nondonors, but the 2 groups had similar blood hematocrits and blood hemoglobin concentrations. Blood donation is simultaneously associated with increased insulin sensitivity and decreased iron stores. Stored iron seems to impact negatively on insulin action even in healthy people, and not just in classic pathologic conditions associated with iron overload (hemochromatosis and hemosiderosis). According to these observations, it is imperative that a definition of excessive iron stores in healthy people be formulated.

  10. Combined cell index in assessing blood donor iron stores.

    PubMed

    Vuk, T; Bingulac-Popović, J; Očić, T; Mayer, L J; Milošević, M; Jukić, I

    2017-02-01

    The aim of this study was to assess the appropriateness of using combined cell index (CCI) in the assessment of iron stores in blood donors. This index is calculated by the formula: red blood cell distribution width (RDW) × 10(4) × mean corpuscular volume (MCV)(-1) × mean corpuscular haemoglobin (MCH)(-1) . Ferritin measurement is a reliable method for estimating iron stores in blood donors. The sensitivity of red blood cell (RBC) parameters of complete blood count in detecting non-anaemic iron deficiency is significantly lower. Consequently, there were several attempts to increase the detection sensitivity by combining these parameters in different indices. This study included 1084 male and 792 female whole blood donors accepted for blood donation. For six RBC parameters with the highest level of correlation relative to ferritin [Hgb, MCV, MCH, mean corpuscular haemoglobin concentration (MCHC), RDW and CCI], diagnostic efficacy in the detection of iron depletion (ferritin <12 µg L(-1) ) was assessed using receiver operating characteristic (ROC) analysis. CCI showed the highest degree of correlation with ferritin (r = -0·373 for men and r = -0·590 for women) and the highest area under the curve (0·961 for men and 0·864 for women). Using the cut-off value of 52·6 for men and 50·6 for women, the corresponding Youden index was the highest for CCI in both genders (0·851 for men and 0·612 for women). The sensitivity and specificity of CCI in the population of male donors were higher in comparison to female donors (0·941 and 0·910 vs 0·851 and 0·761, respectively). Study results confirmed the satisfactory diagnostic value of CCI in detecting depleted iron stores in blood donors. © 2016 British Blood Transfusion Society.

  11. Evaluation of iron stores in blood donors by serum ferritin.

    PubMed

    Mittal, Romilla; Marwaha, Neelam; Basu, Sabita; Mohan, Harsh; Ravi Kumar, A

    2006-12-01

    Regular blood donation can lead to pre-clinical iron deficiency as well as iron deficiency anaemia. There is a need to increase the national voluntary blood donation for safe blood supply. However, there is paucity of data in the country regarding impact of regular voluntary blood donation on iron status of donors. Hence, iron stores were evaluated by serum ferritin estimation in the voluntary blood donors at Chandigarh. 400 voluntary blood donors included in the study were divided into four groups depending upon their periodicity of blood donations. Pre-donation haemoglobin assessment was done by copper sulphate method. Serum ferritin was estimated by indirect ELISA. The number of female donors with deficient iron stores was more as compared to male donors. First time donors had higher mean serum ferritin levels than that in repeat donors. The frequency of donations per year was more predictive of decreased iron stores rather than the number of lifetime donations. An increase in donation frequency was accompanied by a significant decrease in serum ferritin; values <15 microg/l were found in 21 and 46 per cent of male and female donors respectively who donated once per year, in 29 and 27 per cent in those who donated twice per year and in 49 and 100 per cent in those who donated thrice per year. Haemoglobin estimation alone in regular blood donors may not be adequate; serum ferritin estimations may need to be done to detect pre-clinical iron deficiency states. Also, iron supplementation needs to be considered in regular, repeat voluntary blood donors.

  12. Vitamin E nanoemulsion activity on stored red blood cells.

    PubMed

    Silva, C A L; Azevedo Filho, C A; Pereira, G; Silva, D C N; Castro, M C A B; Almeida, A F; Lucena, S C A; Santos, B S; Barjas-Castro, M L; Fontes, A

    2017-06-01

    Stored red blood cells (RBCs) undergo numerous changes that have been termed RBC storage lesion, which can be related to oxidative damage. Vitamin E is an important antioxidant, acting on cell lipids. Thus, this study aimed to investigate vitamin E activity on stored RBCs. We prepared a vitamin E nanoemulsion that was added to RBC units and stored at 4 °C. Controls, without vitamin E, were kept under the same conditions. Reactive oxygen species (ROS) production was monitored for up to 35 days of storage. RBC elasticity was also evaluated using an optical tweezer system. Vitamin E-treated samples presented a significant decrease in ROS production. Additionally, the elastic constant for vitamin E-treated RBCs did not differ from the control. Vitamin E decreased the amount of ROS in stored RBCs. Because vitamin E acts on lipid oxidation, results suggest that protein oxidation should also be considered a key factor for erythrocyte elastic properties. Thus, further studies combining vitamin E with protein antioxidants deserve attention, aiming to better preserve overall stored RBC properties. © 2017 British Blood Transfusion Society.

  13. Procoagulant activity in stored units of red blood cells.

    PubMed

    Aleshnick, Maya; Foley, Jonathan H; Keating, Friederike K; Butenas, Saulius

    2016-06-10

    The procoagulant activity (PA) of stored units of red blood cells (RBC) increases over time, which is related to the expression/exposure of tissue factor (TF). However, there is a discrepancy between the TF measured and changes in PA observed, suggesting that other blood components contribute to this activity. Our goal was to evaluate changes in PA of stored RBCs and to determine possible contributors to it. RBC units from 4 healthy donors were prepared and stored at 4 °C. On selected days, RBC aliquots were reconstituted with autologous plasma and tested in the thromboelastography assay. Corresponding supernatants were tested in a clotting assay. For all donors, the clotting time (CT) of reconstituted RBC units decreased from ∼3000-4000s on day 1 to ∼1000-1600s on day 30, with the most dramatic changes occurring between days 1 and 5. Anti-TF antibody slightly prolonged the CT. The concentration of TF did not change significantly over time and was within the range of 0.3-2.3 pM. Bovine lactadherin (LTD) prolonged the CT of the RBC (by 2.4-3.4-fold in days 3-5 and by 1.3-1.8-fold at day 30). Anti-TF antibody together with LTD had a cumulative effect on the CT prolongation. CT of supernatants responded to both anti-TF and anti-FXIa antibodies. Three contributors to the PA of stored RBC were identified, i.e. FXIa in solution and phosphatidylserine and TF exposed on blood cells and microparticles. Failure of LTD and antibodies to completely eliminate PA suggests that other components of blood could contribute to it. Copyright © 2016 Elsevier Inc. All rights reserved.

  14. Sodium and potassium changes in blood bank stored human erythrocytes.

    PubMed

    Wallas, C H

    1979-01-01

    Storage of red cells for three weeks at 4 C under blood bank conditions resulted in a rise in intracellular Na+ and a fall in intracellular K+ with concomitant opposite changes in Na+ and K+ levels in the suspending plasma. A decline in red blood cell ATP during the storage period did not appear to be contributing to the changes. Increasing red blood cell ATP to levels 2 to 3 times normal did not prevent the cation changes from occurring. When assayed at 37 C in the presence of added Mg++, ouabain-sensitive membrane ATPase activity and kinetics of activation by Na+ were unaffected by the three week period of cold storage. However, when assayed at 4 C without added Mg++, simulating the conditions of storage, ATPase activity was negligible. Sodium and potassium did not change when red blood cells with normal ATP content were stored at 20 to 24 C even in the absence of added Mg++. Thus, a major cause for the development of cation changes in the red blood cell during blood bank storage in the temperature which inhibits membrane ATPase, allowing cations to leak unopposed into and out of the red blood cells.

  15. Mechanical properties of stored red blood cells using optical tweezers

    NASA Astrophysics Data System (ADS)

    Fontes, Adriana; Alexandre de Thomaz, Andre; de Ysasa Pozzo, Liliana; de Lourdes Barjas-Castro, Maria; Brandao, Marcelo M.; Saad, Sara T. O.; Barbosa, Luiz Carlos; Cesar, Carlos Lenz

    2005-08-01

    We have developed a method for measuring the red blood cell (RBC) membrane overall elasticity μ by measuring the deformation of the cells when dragged at a constant velocity through a plasma fluid by an optical tweezers. The deformability of erythrocytes is a critical determinant of blood flow in the microcirculation. We tested our method and hydrodynamic models, which included the presence of two walls, by measuring the RBC deformation as a function of drag velocity and of the distance to the walls. The capability and sensitivity of this method can be evaluated by its application to a variety of studies, such as, the measurement of RBC elasticity of sickle cell anemia patients comparing homozygous (HbSS), including patients taking hydroxyrea (HU) and heterozygous (HbAS) with normal donors and the RBC elasticity measurement of gamma irradiated stored blood for transfusion to immunosupressed patients as a function of time and dose. These studies show that the technique has the sensitivity to discriminate heterozygous and homozygous sickle cell anemia patients from normal donors and even follow the course of HU treatment of Homozygous patients. The gamma irradiation studies show that there is no significant change in RBC elasticity over time for up to 14 days of storage, regardless of whether the unit was irradiated or not, but there was a huge change in the measured elasticity for the RBC units stored for more than 21 days after irradiation. These finds are important for the assessment of stored irradiated RBC viability for transfusion purposes because the present protocol consider 28 storage days after irradiation as the limit for the RBC usage.

  16. The heritability of hemolysis in stored human red blood cells

    PubMed Central

    van ‘t Erve, Thomas J.; Wagner, Brett A.; Martin, Sean M.; Knudson, C. Michael; Blendowski, Robyn; Keaton, Mignon; Holt, Tracy; Hess, John R.; Buettner, Garry R.; Ryckman, Kelli K.; Darbro, Benjamin W.; Murray, Jeffrey C.; Raife, Thomas J.

    2015-01-01

    Background The transfusion of red blood cells (RBCs) with maximum therapeutic efficacy is a major goal in transfusion medicine. One of the criteria used in determining stored RBC quality is end of storage hemolysis. Between donors, a wide range of hemolysis is observed under identical storage conditions. Here, a potential mechanism for this wide range is investigated. We hypothesize that the magnitude of hemolysis is a heritable trait. Also, we investigated correlations between hemolysis and RBC metabolites; this will establish pathways influencing hemolysis as future targets for genetic analysis. Study Design and Methods Units of RBCs from identical and non-identical twins were collected and stored under standard conditions for 56 days. Hemolysis, ATP, and total glutathione were measured throughout storage. Non-targeted metabolic analyses were performed on RBCs that had been stored for 28 days. Heritability was determined by comparing values between identical and non-identical twins. Results Hemolysis was found to be heritable (mean >45 %) throughout the storage period. Correlations were observed between hemolysis and metabolites from the amino acid, sugar, and purine metabolism, lipid metabolism and transport, and glycolysis pathways. Three metabolites also exhibited heritability (> 20%). No correlation was found with ATP or total glutathione. Conclusion The susceptibility of RBCs to lysis during storage is partly determined by inheritance. We have also uncovered several pathways that are candidate targets for future genome wide association studies. These findings will aid in the design of better storage solutions and the development of donor screening tools that minimize hemolysis during storage. PMID:25644965

  17. Inactivation of human immunodeficiency virus type 1 in blood samples stored as high-salt lysates.

    PubMed

    Zolg, J W; Lanciotti, R S; Wendlinger, M; Meyer, W A

    1990-09-01

    Blood samples to be tested for the presence of parasite DNA by using specific DNA probes are routinely stored in our laboratory as high-salt lysates (HSL). To safeguard against the risk of accidental infection with etiological agents such as the human immunodeficiency virus type 1 (HIV-1) while manipulating large numbers of blood samples in preparation for DNA probing, we determined the residual infectivity of HIV-1 after exposure to HSL components. Both high-titer virus stocks or provirus-carrying cells, suspended either in tissue culture medium or freshly drawn blood, were completely inactivated upon contact with the HSL components. This was verified by the absence of any detectable HIV-1-specific antigen in the supernatants of long-term cultures and the absence of virus-specific DNA fragments after amplification by polymerase chain reaction with DNA from such cultures as target DNA. These results support the conclusion that the virus is in fact completely inactivated by contact with the HSL components, rendering blood specimens stored as HSL noninfectious in regard to HIV-1.

  18. Raman spectroscopy of stored red blood cells: evaluating clinically-relevant biochemical markers in donated blood

    NASA Astrophysics Data System (ADS)

    Atkins, Chad G.; Buckley, Kevin; Chen, Deborah; Schulze, H. G.; Devine, Dana V.; Blades, Michael W.; Turner, Robin F. B.

    2015-07-01

    Modern transfusion medicine relies on the safe, secure, and cost-effective delivery of donated red blood cells (RBCs). Once isolated, RBCs are suspended in a defined additive solution and stored in plastic blood bags in which, over time, they undergo chemical, physiological, and morphological changes that may have a deleterious impact on some patients. Regulations limit the storage period to 42 days and the cells do not routinely undergo analytical testing before use. In this study, we use Raman spectroscopy to interrogate stored RBCs and we identify metabolic and cell-breakdown products, such as haemoglobin and membrane fragments, that build-up in the blood bags as the cells age. Our work points the way to the development of an instrument which could quickly and easily assess the biochemical nature of stored RBC units before they are transfused.

  19. Quality of red blood cells isolated from umbilical cord blood stored at room temperature.

    PubMed

    Zhurova, Mariia; Akabutu, John; Acker, Jason

    2012-01-01

    Red blood cells (RBCs) from cord blood contain fetal hemoglobin that is predominant in newborns and, therefore, may be more appropriate for neonatal transfusions than currently transfused adult RBCs. Post-collection, cord blood can be stored at room temperature for several days before it is processed for stem cells isolation, with little known about how these conditions affect currently discarded RBCs. The present study examined the effect of the duration cord blood spent at room temperature and other cord blood characteristics on cord RBC quality. RBCs were tested immediately after their isolation from cord blood using a broad panel of quality assays. No significant decrease in cord RBC quality was observed during the first 65 hours of storage at room temperature. The ratio of cord blood to anticoagulant was associated with RBC quality and needs to be optimized in future. This knowledge will assist in future development of cord RBC transfusion product.

  20. Quality of Red Blood Cells Isolated from Umbilical Cord Blood Stored at Room Temperature

    PubMed Central

    Zhurova, Mariia; Akabutu, John; Acker, Jason

    2012-01-01

    Red blood cells (RBCs) from cord blood contain fetal hemoglobin that is predominant in newborns and, therefore, may be more appropriate for neonatal transfusions than currently transfused adult RBCs. Post-collection, cord blood can be stored at room temperature for several days before it is processed for stem cells isolation, with little known about how these conditions affect currently discarded RBCs. The present study examined the effect of the duration cord blood spent at room temperature and other cord blood characteristics on cord RBC quality. RBCs were tested immediately after their isolation from cord blood using a broad panel of quality assays. No significant decrease in cord RBC quality was observed during the first 65 hours of storage at room temperature. The ratio of cord blood to anticoagulant was associated with RBC quality and needs to be optimized in future. This knowledge will assist in future development of cord RBC transfusion product. PMID:24089645

  1. Technical improvements in culturing blood.

    PubMed

    Pardini, Giacomo

    2015-01-01

    Blood culture is a laboratory test where a blood specimen, taken from a patient, is inoculated into bottles containing culture media to determine if infection-causing microorganisms (bacteria or fungi) have invaded the patient's bloodstream. This test is an important investigation with major implications for the diagnosis and treatment of patients with bloodstream infections and possible sepsis. Moreover, blood culture will also provide the etiologic agent for antimicrobial susceptibility testing, enabling optimization of antibiotic therapy with significant impact on the outcome of the disease. Even if the potential benefices of blood culture are well known, critical factors mainly in pre- and post-analytical phases can reduce the clinical value of this test.

  2. Guidance for storing blood samples in laboratories performing complete blood count with differential.

    PubMed

    Cornet, E; Behier, C; Troussard, X

    2012-12-01

    The complete blood count (CBC) with differential leukocyte count (DIFF) is an important part of clinical laboratory analyses and provides crucial data for clinicians. Delivery time after blood collection and conditions of storage is known to affect the reliability of results of some hematologic parameters. The aim of this study was to assess the variations of hematologic parameters over time and the influence of storage temperature. Blood samples were randomly selected from hospitalized patients and stored at room temperature and at 4 °C. CBC and DIFF were performed on an automated hematology analyzer and the results between the two groups were compared. Samples stored at room temperature showed an important increase in mean corpuscular volume and hematocrit and a decrease in mean corpuscular hemoglobin concentration. Neutrophil counts tended to increase, whereas monocyte counts tended to decrease. Storing samples at 4 °C improved reproducibility over time of all quantitative and qualitative parameters. We also observed that NEUT-X, a routine parameter useful in detecting myelodysplastic syndrome, became unreliable when analyzed 24 h after sample collection. Our results led us to recommend that samples should be analyzed within 6 h, particularly if samples are transported at room temperature. We also recommend storing samples at 4 °C in case of remote CBC analysis, especially in the context of clinical trials. © 2012 Blackwell Publishing Ltd.

  3. Non-invasive spectroscopy of transfusable red blood cells stored inside sealed plastic blood-bags.

    PubMed

    Buckley, K; Atkins, C G; Chen, D; Schulze, H G; Devine, D V; Blades, M W; Turner, R F B

    2016-03-07

    After being separated from (donated) whole blood, red blood cells are suspended in specially formulated additive solutions and stored (at 4 °C) in polyvinyl chloride (PVC) blood-bags until they are needed for transfusion. With time, the prepared red cell concentrate (RCC) is known to undergo biochemical changes that lower effectiveness of the transfusion, and thus regulations are in place that limit the storage period to 42 days. At present, RCC is not subjected to analytical testing prior to transfusion. In this study, we use Spatially Offset Raman Spectroscopy (SORS) to probe, non-invasively, the biochemistry of RCC inside sealed blood-bags. The retrieved spectra compare well with conventional Raman spectra (of sampled aliquots) and are dominated by features associated with hemoglobin. In addition to the analytical demonstration that SORS can be used to retrieve RCC spectra from standard clinical blood-bags without breaking the sterility of the system, the data reveal interesting detail about the oxygenation-state of the stored cells themselves, namely that some blood-bags unexpectedly contain measurable amounts of deoxygenated hemoglobin after weeks of storage. The demonstration that chemical information can be obtained non-invasively using spectroscopy will enable new studies of RCC degeneration, and points the way to a Raman-based instrument for quality-control in a blood-bank or hospital setting.

  4. Storage of Blood Components Does Not Decrease Haemostatic Potential: In vitro Assessment of Fresh versus Stored Blood Components Using Thromboelastography.

    PubMed

    Bartfeld, Galia; Ellis, Martin; Lubetzky, Aharon; Yahalom, Vered; Kenet, Gili

    2010-01-01

    SUMMARY: BACKGROUND: Major surgery and severe trauma typically lead to massive blood loss requiring rapid transfusion of large amounts of blood products. It has been suggested that fresh, unrefrigerated whole blood provides a haemostatic advantage in this setting. The aim of the current study was to compare the clot formation parameters of fresh, unrefrigerated whole blood and whole blood reconstituted from components stored for varying periods of time, using rotational thromboelastography (ROTEM®). METHODS: Fresh whole blood and reconstituted whole blood using combinations of non-leucoreduced red cell units (stored for 7, 14, 21, 28, or 35 days), platelet concentrates (stored for 1, 3 or 5 days), and fresh frozen plasma (stored for 6 months) were analysed using ROTEM. Measurements of the clotting time (CT), clot formation time (CFT), and maximal clot firmness (MCF) were compared between units of fresh whole blood and reconstituted whole blood samples. RESULTS: There was no difference in the haemostatic parameters measured of fresh whole blood and reconstituted whole blood using red cell units stored for less than 21 days. ROTEM demonstrated that the CT and CFT were significantly shorter for reconstituted whole blood samples using red cells stored for longer than 21 days when compared to fresh whole blood and to reconstituted whole blood samples using red cell units stored for less than 21 days. The CT was inversely correlated to the duration of platelet storage. The MCF was unchanged regardless of duration of blood product storage. CONCLUSION: Fresh unrefrigerated whole blood and blood products stored for short duration (less than 21 days) were not superior to those stored for longer durations.

  5. Storage of Blood Components Does Not Decrease Haemostatic Potential: In vitro Assessment of Fresh versus Stored Blood Components Using Thromboelastography

    PubMed Central

    Bartfeld, Galia; Ellis, Martin; Lubetzky, Aharon; Yahalom, Vered; Kenet, Gili

    2010-01-01

    Summary Background Major surgery and severe trauma typically lead to massive blood loss requiring rapid transfusion of large amounts of blood products. It has been suggested that fresh, unrefrigerated whole blood provides a haemostatic advantage in this setting. The aim of the current study was to compare the clot formation parameters of fresh, unrefrigerated whole blood and whole blood reconstituted from components stored for varying periods of time, using rotational thromboelastography (ROTEM®). Methods Fresh whole blood and reconstituted whole blood using combinations of non-leucoreduced red cell units (stored for 7, 14, 21, 28, or 35 days), platelet concentrates (stored for 1, 3 or 5 days), and fresh frozen plasma (stored for 6 months) were analysed using ROTEM. Measurements of the clotting time (CT), clot formation time (CFT), and maximal clot firmness (MCF) were compared between units of fresh whole blood and reconstituted whole blood samples. Results There was no difference in the haemostatic parameters measured of fresh whole blood and reconstituted whole blood using red cell units stored for less than 21 days. ROTEM demonstrated that the CT and CFT were significantly shorter for reconstituted whole blood samples using red cells stored for longer than 21 days when compared to fresh whole blood and to reconstituted whole blood samples using red cell units stored for less than 21 days. The CT was inversely correlated to the duration of platelet storage. The MCF was unchanged regardless of duration of blood product storage. Conclusion Fresh unrefrigerated whole blood and blood products stored for short duration (less than 21 days) were not superior to those stored for longer durations. PMID:21416027

  6. Inhibition of Platelet Aggregation by Supernates from Stored Red Blood Cells

    DTIC Science & Technology

    2010-04-01

    platelet aggregates in fresh whole blood.[10] In those experiments , we observed that platelets in blood incubated with supernates from stored RBC...volumes in sterile cryovials, and the vials were stored at -80C. For each experiment , aliquots were thawed quickly in a 37°C water bath just before...The final ratio of whole blood to supernate was 2:1. For each experiment , blood was collected first into one red top Vacutainer tube (no anti

  7. Temperature-dependent haemolytic propensity of CPDA-1 stored red blood cells vs whole blood - Red cell fragility as donor signature on blood units.

    PubMed

    Tzounakas, Vassilis L; Anastasiadi, Alkmini T; Karadimas, Dimitrios G; Zeqo, Redisa A; Georgatzakou, Hara T; Pappa, Olga D; Papatzitze, Olga A; Stamoulis, Konstantinos E; Papassideri, Issidora S; Antonelou, Marianna H; Kriebardis, Anastasios G

    2017-09-01

    To preserve cellular integrity and avoid bacterial growth, storage and transfer of blood and blood products follow strict guidelines in terms of temperature control. We evaluated the impact of ineligible warming of whole blood donations on the quality of blood components. One-hundred and twenty units of whole blood (WB) from eligible blood donors were collected in CPDA-1 and stored at 4±2 °C. During shipment to the blood processing centre, a gradual warming up to 17 °C was recorded within a period of less than eight hours. The warmed units were processed to packed red blood cells (PRBCs) or stored as WB units at 4±2 °C. In-bag haemolysis, osmotic fragility (mean corpuscular fragility, MCF) and bacterial growth were assessed in blood and blood components throughout the storage period. Normal basal and early storage levels of haemolysis were recorded in both PRBC and WB units. Thereafter, PRBCs exhibited higher average in-bag haemolysis and MCF index compared to the WB units throughout the storage. Moreover, 14.3 and 52.4% of the PRBC units exceeded the upper permissible limit of 0.8% haemolysis at the middle (1.220±0.269%) or late (1.754±0.866%) storage period, respectively. MCF index was similar in all PRBCs at the middle of storage but significantly lower in the non-haemolysed compared to the haemolysed units of PRBCs on the last days. The fragility of stored RBCs was proportional to the donor-related values of day 2 samples (r=0.861, p<10(-32)). In the qualified PRBCs, MCF was correlated with haemolysis at every time point of the storage period (r=0.332, p<0.050). Bacterial growth was detected by blood culture in two units of PRBCs. Transient, gradient warming of whole blood from 4 to 17 °C led to increased incidence of in-bag haemolysis in PRBC but not in WB units. Haemolysis is a multi-parametric phenotype of stored blood, and MCF is a donor-related and highly dynamic measure that can, in part, predict the storage lesion.

  8. A National Coordinating Center for Prehospital Trauma Research Funding Transfusion Using Stored Fresh Whole Blood

    DTIC Science & Technology

    2016-09-01

    Using Stored Fresh Whole Blood PRINCIPAL INVESTIGATOR: Donald Jenkins, M.D. CONTRACTING ORGANIZATION: NATIONAL TRAUMA INSTITUTE San Antonio...Std. Z39.18 A National Coordinating Center for Prehospital Trauma Research Funding Transfusion Using Stored Fresh Whole Blood Table of Contents...Warfighter Medical Research Program Funding to extend the work previously completed looking at the use of fresh whole blood FWB) and its ability to

  9. Comparison of Stored Umbilical Cord Blood and Adult Donor Blood: Transfusion Feasibility

    PubMed Central

    Tokan, Rola Sahyoun; Arsan, Saadet; Erdeve, Ömer; Solaz, Nuri; Avcı, Aslıhan; Ülkar, Serenay Elgün; Gülyapar, Elif; Üstünyurt, Zeynep; Bıyıklı, Zeynep; Kemahlı, Sabri

    2012-01-01

    Objective: This study aimed to compare the storage properties of red blood cell (RBC) concentrates of umbilical cordblood (UCB) and adult donor blood (ADB), and to evaluate the feasibility of UCB-RBC concentrate as an autologoussource for blood transfusion in very low birth weight (VLBW) preterm neonates. Material and Methods: In all, 30 newborn (10 preterm, 20 full term) UCB and 31 ADB units were collected.RBC concentrates were stored and compared with regard to pH, potassium (K+), 2,3-biphosphoglycerate (2-3-BPG),adenosine tri-phosphate (ATP), plasma Hb, and bacterial contamination on d 1, 21, and 35 of storage. Results: The K+ level increased with time and differed significantly between storage d 1 and 21, and between storaged 1 and 35 in both the UCB and ADB units. Initial and d 21 K+ levels were higher in the UCB units than in the ADBunits. The 2,3-BPG level did not differ significantly between the UCB-PRC and ADB-PRC samples. After 35 d of storageboth UCB-PRC and ADB-PRC samples exhibited significant differences from the initial free Hb, intracellular ATP, andpH values. Significant differences in intracellular ATP and pH were also observed between the UCB-PRC and ADB-PRCsamples. Conclusion: The volume of harvested and prepared UCB-PRC can be used for some of the blood transfusions requiredduring the neonatal period and thus may decrease the number of allogeneic transfusions, especially in preterm newborns.The hematological and biochemical changes that occurred in UCB during storage were comparable with those observedin ADB, and do not pose a risk to the immature metabolism of neonates. UCB-RPC prepared and stored under standardconditions can be a safe alternative RBC source for transfusions in VLBW newborns. PMID:24744666

  10. Comparison of stored umbilical cord blood and adult donor blood: transfusion feasibility.

    PubMed

    Tokan, Rola Sahyoun; Arsan, Saadet; Erdeve, Omer; Solaz, Nuri; Avcı, Aslıhan; Ulkar, Serenay Elgün; Gülyapar, Elif; Ustünyurt, Zeynep; Bıyıklı, Zeynep; Kemahlı, Sabri

    2012-09-01

    This study aimed to compare the storage properties of red blood cell (RBC) concentrates of umbilical cordblood (UCB) and adult donor blood (ADB), and to evaluate the feasibility of UCB-RBC concentrate as an autologoussource for blood transfusion in very low birth weight (VLBW) preterm neonates. In all, 30 newborn (10 preterm, 20 full term) UCB and 31 ADB units were collected.RBC concentrates were stored and compared with regard to pH, potassium (K(+)), 2,3-biphosphoglycerate (2-3-BPG),adenosine tri-phosphate (ATP), plasma Hb, and bacterial contamination on d 1, 21, and 35 of storage. The K(+) level increased with time and differed significantly between storage d 1 and 21, and between storaged 1 and 35 in both the UCB and ADB units. Initial and d 21 K(+) levels were higher in the UCB units than in the ADBunits. The 2,3-BPG level did not differ significantly between the UCB-PRC and ADB-PRC samples. After 35 d of storageboth UCB-PRC and ADB-PRC samples exhibited significant differences from the initial free Hb, intracellular ATP, andpH values. Significant differences in intracellular ATP and pH were also observed between the UCB-PRC and ADB-PRCsamples. The volume of harvested and prepared UCB-PRC can be used for some of the blood transfusions requiredduring the neonatal period and thus may decrease the number of allogeneic transfusions, especially in preterm newborns.The hematological and biochemical changes that occurred in UCB during storage were comparable with those observedin ADB, and do not pose a risk to the immature metabolism of neonates. UCB-RPC prepared and stored under standardconditions can be a safe alternative RBC source for transfusions in VLBW newborns.

  11. Assessment of viable bacteria and bacterial DNA in blood and bloodstain specimens stored under various conditions.

    PubMed

    Hosokawa-Muto, Junji; Fujinami, Yoshihito; Mizuno, Natsuko

    2013-11-01

    Microbial forensic specimens that are collected at biocrime and bioterrorism scenes include blood, tissue, cloths containing biological fluids, swabs, water, soil, and aerosols. It is preferable that pathogens in such specimens are alive and kept in a steady state. Specimens may be stored for a prolonged period before analysis; therefore, it is important to understand the effect of the storage conditions on the pathogens contained within the specimens. In this study, we prepared blood and bloodstain specimens containing Gram-negative or -positive bacteria, stored the samples for 482 days under various conditions, and measured viable bacterial counts and total bacterial contents in the samples. Viable bacteria were preserved well in the samples stored at -30 and -80 °C, but were diminished or undetectable in the samples stored at 4 °C and room temperature. The total bacterial content was maintained in the blood samples stored at -30 and -80 °C and in the bloodstain samples stored under all temperature conditions, but decreased in the blood samples stored at 4 °C and room temperature. This study showed that the storage conditions affected viable bacteria and bacterial DNA and that freezing and drying were significant for their long-term storage. We provide important information for the storage of microbial forensic specimens.

  12. Modeling the potential impact on the US blood supply of transfusing critically ill patients with fresher stored red blood cells.

    PubMed

    Simonetti, Arianna; Ezzeldin, Hussein; Menis, Mikhail; McKean, Stephen; Izurieta, Hector; Anderson, Steven A; Forshee, Richard A

    2017-01-01

    Although some studies have suggested that transfusion recipients may have better medical outcomes if transfused with red blood cell units stored for a short time, the overall body of evidence shows mixed results. It is important to understand how using fresher stored red blood cell units for certain patient groups may affect blood availability. Based on the Stock-and-Flow simulation model of the US blood supply developed by Simonetti et al. 2014, we evaluated a newly implemented allocation method of preferentially transfusing fresher stored red blood cell units to a subset of high-risk group of critically ill patients and its potential impact on supply. Simulation results showed that, depending on the scenario, the US blood total supply might be reduced between 2-42%, when compared to the standard of care in transfusion medicine practice. Among our simulated scenarios, we observed that the number of expired red blood cell units modulated the supply levels. The age threshold of the required red blood cell units was inversely correlated with both the supply levels and the number of transfused units that failed to meet that age threshold. To our knowledge, this study represents the first attempt to develop a comprehensive framework to evaluate the impact of preferentially transfusing fresher stored red blood cells to the higher-risk critically ill patients on supply. Model results show the difficulties to identify an optimal scenario.

  13. Modeling the potential impact on the US blood supply of transfusing critically ill patients with fresher stored red blood cells

    PubMed Central

    Ezzeldin, Hussein; Menis, Mikhail; McKean, Stephen; Izurieta, Hector; Anderson, Steven A.; Forshee, Richard A.

    2017-01-01

    Background Although some studies have suggested that transfusion recipients may have better medical outcomes if transfused with red blood cell units stored for a short time, the overall body of evidence shows mixed results. It is important to understand how using fresher stored red blood cell units for certain patient groups may affect blood availability. Methods Based on the Stock-and-Flow simulation model of the US blood supply developed by Simonetti et al. 2014, we evaluated a newly implemented allocation method of preferentially transfusing fresher stored red blood cell units to a subset of high-risk group of critically ill patients and its potential impact on supply. Results Simulation results showed that, depending on the scenario, the US blood total supply might be reduced between 2-42%, when compared to the standard of care in transfusion medicine practice. Among our simulated scenarios, we observed that the number of expired red blood cell units modulated the supply levels. The age threshold of the required red blood cell units was inversely correlated with both the supply levels and the number of transfused units that failed to meet that age threshold. Conclusion To our knowledge, this study represents the first attempt to develop a comprehensive framework to evaluate the impact of preferentially transfusing fresher stored red blood cells to the higher-risk critically ill patients on supply. Model results show the difficulties to identify an optimal scenario. PMID:28319164

  14. Impact of the age of stored blood on trauma patient mortality: a systematic review

    PubMed Central

    Sowers, Nicholas; Froese, Patrick C.; Erdogan, Mete; Green, Robert S.

    2015-01-01

    Background The impact of the age of stored red blood cells on mortality in patients sustaining traumatic injuries requiring transfusion of blood products is unknown. The objective of this systematic review was to identify and describe the available literature on the use of older versus newer blood in trauma patient populations. Methods We searched PubMed, Embase, Lilac and the Cochrane Database for published studies comparing the transfusion of newer versus older red blood cells in adult patients sustaining traumatic injuries. Studies included for review reported on trauma patients receiving transfusions of packed red blood cells, identified the age of stored blood that was transfused and reported patient mortality as an end point. We extracted data using a standardized form and assessed study quality using the Newcastle–Ottawa Scale. Results Seven studies were identified (6780 patients) from 3936 initial search results. Four studies reported that transfusion of older blood was independently associated with increased mortality in trauma patients, while 3 studies did not observe any increase in patient mortality with the use of older versus newer blood. Three studies associated the transfusion of older blood with adverse patient outcomes, including longer stay in the intensive care unit, complicated sepsis, pneumonia and renal dysfunction. Studies varied considerably in design, volumes of blood transfused and definitions applied for old and new blood. Conclusion The impact of the age of stored packed red blood cells on mortality in trauma patients is inconclusive. Future investigations are warranted. PMID:26384149

  15. Mechanism of faster NO scavenging by older stored red blood cells.

    PubMed

    Liu, Chen; Liu, Xiaohua; Janes, John; Stapley, Ryan; Patel, Rakesh P; Gladwin, Mark T; Kim-Shapiro, Daniel B

    2014-01-01

    The blood storage lesion involves morphological and biochemical changes of red blood cells (RBCs) that occur during storage. These include conversion of the biconcave disc morphology to a spherical one, decreased mean corpuscular hemoglobin concentration, varied mean corpuscular volume, reduced integrity of the erythrocyte membrane with formation of microparticles, and increased cell-free hemoglobin. We studied the extent that older stored red blood cells scavenge nitric oxide (NO) faster than fresher stored red blood cells. Using electron paramagnetic resonance spectroscopy and stopped-flow absorption spectroscopy to measure the rate of NO uptake and reaction with hemoglobin in red cells, we found that older stored red blood cells scavenge NO about 1.8 times faster than fresher ones. Based on these experimental data, we simulated NO scavenging by fresher or older stored red blood cells with a biconcave or spherical geometry, respectively, in order to explore the mechanism of NO scavenging related to changes that occur during blood storage. We found that red blood cells with a spherical geometry scavenges NO about 2 times slower than ones with a biconcave geometry, and a smaller RBC hemoglobin concentration or volume increases NO scavenging by red blood cells. Our simulations demonstrate that even the most extreme possible changes in mean corpuscular hemoglobin concentration and mean corpuscular volume that favor increased NO scavenging are insufficient to account for what is observed experimentally. Therefore, RBC membrane permeability must increase during storage and we find that the permeability is likely to increase between 5 and 70 fold. Simulations using a two-dimensional blood vessel show that even a 5-fold increase in membrane permeability to NO can reduce NO bioavailability at the smooth muscle. Transfusion of older stored blood may be harmful. Older stored red blood cells scavenge nitric oxide more than fresher cells. As stored red blood cells age

  16. Pulmonary Hypertension in Lambs Transfused with Stored Blood is Prevented by Breathing Nitric Oxide

    PubMed Central

    Baron, David M.; Yu, Binglan; Lei, Chong; Bagchi, Aranya; Beloiartsev, Arkadi; Stowell, Christopher P.; Steinbicker, Andrea U.; Malhotra, Rajeev; Bloch, Kenneth D.; Zapol, Warren M.

    2012-01-01

    Background During extended storage, erythrocytes undergo functional changes. These changes reduce the viability of erythrocytes leading to release of oxyhemoglobin, a potent scavenger of nitric oxide. We hypothesized that transfusion of ovine packed erythrocytes (PRBC) stored for prolonged periods would induce pulmonary vasoconstriction in lambs, and that reduced vascular nitric oxide concentrations would increase this vasoconstrictor effect. Methods We developed a model of autologous stored blood transfusion in lambs (n=36). Leukoreduced blood was stored for either 2 days (fresh PRBC) or 40 days (stored PRBC). Fresh or stored PRBC were transfused into donors instrumented for awake hemodynamic measurements. Hemodynamic effects of PRBC transfusion were also studied after infusion of NG-nitro-L-arginine methyl-ester (25 mg/kg) or during inhalation of nitric oxide (80 ppm). Results Cell-free hemoglobin levels were higher in the supernatant of stored PRBC than in supernatant of fresh PRBC (Mean±SD, 148±20 versus 41±13 mg/dl, respectively, P<0.001). Pulmonary artery pressure during transfusion of stored PRBC transiently increased from 13±1 to 18±1 mmHg (P<0.001) and was associated with increased plasma hemoglobin concentrations. NG-nitro-L-arginine methyl-ester potentiated the increase in pulmonary arterial pressure induced by transfusing stored PRBC, whereas inhalation of nitric oxide prevented the vasoconstrictor response. Conclusions Our results suggest that patients with reduced vascular nitric oxide levels due to endothelial dysfunction may be more susceptible to adverse effects of transfusing blood stored for prolonged periods. These patients might benefit from transfusion of fresh PRBC, when available, or inhaled nitric oxide supplementation to prevent the pulmonary hypertension associated with transfusion of stored PRBC. PMID:22293717

  17. Isolation of Salmonella typhi from Standard Whole Blood Culture versus Blood-Clot Cultures

    DTIC Science & Technology

    1988-12-01

    The use of 10% oxgall and bile broth medium, both supplemented with freshly prepared 100 u/ml streptokinase, for isolating Salmonella typhi by clot...significantly better rate of isolation than the clot culture methods. Keywords: Cultures biology; Clot cultures; Salmonella typhi ; Isolation of S. typhi; Whole blood culture; Blood-clot culture; Reprints.

  18. Platelet-rich fibrin prepared from stored whole-blood samples.

    PubMed

    Isobe, Kazushige; Suzuki, Masashi; Watanabe, Taisuke; Kitamura, Yutaka; Suzuki, Taiji; Kawabata, Hideo; Nakamura, Masayuki; Okudera, Toshimitsu; Okudera, Hajime; Uematsu, Kohya; Nakata, Koh; Tanaka, Takaaki; Kawase, Tomoyuki

    2017-12-01

    In regenerative therapy, self-clotted platelet concentrates, such as platelet-rich fibrin (PRF), are generally prepared on-site and are immediately used for treatment. If blood samples or prepared clots can be preserved for several days, their clinical applicability will expand. Here, we prepared PRF from stored whole-blood samples and examined their characteristics. Blood samples were collected from non-smoking, healthy male donors (aged 27-67 years, N = 6), and PRF clots were prepared immediately or after storage for 1-2 days. Fibrin fiber was examined by scanning electron microscopy. Bioactivity was evaluated by means of a bioassay system involving human periosteal cells, whereas PDGF-BB concentrations were determined by an enzyme-linked immunosorbent assay. Addition of optimal amounts of a 10% CaCl2 solution restored the coagulative ability of whole-blood samples that contained an anticoagulant (acid citrate dextrose) and were stored for up to 2 days at ambient temperature. In PRF clots prepared from the stored whole-blood samples, the thickness and cross-links of fibrin fibers were almost identical to those of freshly prepared PRF clots. PDGF-BB concentrations in the PRF extract were significantly lower in stored whole-blood samples than in fresh samples; however, both extracts had similar stimulatory effects on periosteal-cell proliferation. Quality of PRF clots prepared from stored whole-blood samples is not reduced significantly and can be ensured for use in regenerative therapy. Therefore, the proposed method enables a more flexible treatment schedule and choice of a more suitable platelet concentrate immediately before treatment, not after blood collection.

  19. Prevalence of Medical Conditions Potentially Amenable to Cellular Therapy among Families Privately Storing Umbilical Cord Blood.

    PubMed

    Mazonson, Peter; Kane, Mark; Colberg, Kelin; Harris, Heather; Brown, Heather; Mohr, Andrew; Ziman, Alyssa; Santas, Chris

    2017-01-01

    Introduction Little is known about the prevalence of conditions potentially amenable to cellular therapy among families storing umbilical cord blood in private cord blood banks. Methods A cross-sectional study of families with at least one child who stored umbilical cord blood in the largest private cord blood bank in the United States was performed. Respondent families completed a questionnaire to determine whether children with stored cord blood or a first-degree relative had one or more of 16 conditions amenable primarily to allogeneic stem cell transplant ("transplant indications") or 16 conditions under investigation for autologous stem cell infusion ("regenerative indications"), regardless of whether they received a transplant or infusion. Results 94,803 families responded, representing 33.3 % of those surveyed. Of respondent families, 16.01 % indicated at least one specified condition. 1.64 % reported at least one first-degree member with a transplant indication potentially treatable with an allogeneic stem cell transplant. The most common transplant indications reported among first-degree family members were Non-Hodgkin's Lymphoma (0.33 %), Hodgkin's Lymphoma (0.30 %), and Acute Lymphoblastic Leukemia (0.28 %). 4.23 % reported at least one child with a regenerative indication potentially treatable with an autologous stem cell infusion. The most common regenerative indications among children with stored umbilical cord blood were Autism/Autism Spectrum Disorder/Apraxia (1.93 %), Other Developmental Delay (1.36 %), and Congenital Heart Defect (0.87 %). Discussion Among families storing umbilical cord blood in private cord blood banks, conditions for which stem cell transplant or infusion may be indicated, or are under investigation, appear to be prevalent, especially for regenerative medicine indications.

  20. Mechanical property analysis of stored red blood cell using optical tweezers.

    PubMed

    Li, Yanjie; Wen, Cheng; Xie, Huimin; Ye, Anpei; Yin, Yajun

    2009-05-01

    The deformation of human red blood cells subjected to direct stretching by optical tweezers was analyzed. The maximum force exerted by optical tweezers on the cell via a polystyrene microbead 5microm in diameter was 315pN. Digital image correlation (DIC) method was introduced to calculate the force and the deformation of the cell for the first time. Force-extension relation curves of the biconcave cell were quantitatively assessed when erythrocytes were stored in Alsever's Solution for 2 days, 5 days, 7 days and 14 days respectively. Experiment results demonstrated that the deformability of red blood cells was impaired with the stored time.

  1. Platelet concentrates from fresh or overnight-stored blood, an international study.

    PubMed

    Dijkstra-Tiekstra, M J; van der Meer, P F; Cardigan, R; Devine, D; Prowse, C; Sandgren, P; de Wildt-Eggen, J

    2011-01-01

    Whole blood and also buffy coats (BCs) can be held for a few hours or overnight before processing into blood components or platelet concentrates (PCs). Individual studies have reported a range of outcomes regarding in vitro variables for PCs prepared from fresh and stored whole blood. In this multicenter study, effects of storage of whole blood or BCs on the in vitro quality of PCs were studied. The leukoreduced BC PCs were prepared from fresh BCs (2-8 hr after collection; fresh/fresh), from BCs at 20 to 24 hours after collection (fresh/stored), or from BCs prepared from whole blood stored for 20 to 24 hours (stored/fresh). PCs were stored on a flat-bed shaker at 20 to 24°C for 7 days. PCs were tested on Days 0 (only fresh/fresh), 1, 5, and 7 for in vitro quality. There were six participating centers that tested all three conditions with n = 6 per condition. In comparison to fresh/stored and stored/fresh PCs, fresh/fresh PCs exhibited a lower platelet (PLT) count (Day 1-220 × 10(9) ± 70 × 10(9) vs. 324 × 10(9) ± 50 × 10(9) and 368 × 10(9) ± 56 × 10(9) PLTs/PC), lactate, pCO(2), and hypotonic shock response (HSR; Days 5 and 7; Day 7-50 ± 13% vs. 57 ± 12 and 63 ± 11%) and a higher pH, glucose, pO(2), and CD62P expression (than stored/fresh PCs only; Day 7-33 ± 10% vs. 28 ± 12 and 24 ± 11%; p < 0.05). No differences were observed for volume, swirling effect, white blood cell count, annexin V binding, or aggregation between these conditions. Based on PLT count, HSR, and PLT activation, PCs are best prepared after 20 to 24 hours hold of the whole blood or BCs. © 2010 American Association of Blood Banks.

  2. Evaluation of Iron Store by Serum Ferritin in Healthy Blood Donors of Bangladesh.

    PubMed

    Hoque, M M; Adnan, S D; Karim, S; Mamun, M A; Nandy, S; Faruki, M A; Islam, K

    2016-07-01

    Iron stores in the body exist primarily in the form of ferritin. Small amounts of ferritin secreted into the plasma and plasma ferritin is positively correlated with the size of the total body iron stores. The present study conducted to determine the iron status using the serum ferritin level among healthy Bangladeshi blood donors. The present cross sectional study was conducted in the Department of Transfusion Medicine, Dhaka Medical College, Dhaka, Bangladesh from July 2011 to June 2012. Blood donor signed informed consent and has satisfactory pre-donation health assessment and satisfactory post-donation blood test results were included in the study. Full blood counts were performed within 4 hours of collection using an automated haematology analyzer. Serum ferritin was measured using a validated enzyme immunoassay. Data were analyzed using SPSS version 16 (SPPS Incorporation, Chicago, IL, USA). P value <0.05 was considered as statistically significant. Total 100 blood donors were included in the study, among them 88 were male and 12 were female. Mean±SD of the age of the respondents was 26.8±5.9 years with a range of 19 to 45 years. Mean±SD of heamoglobin level (gm/dl) and total count of Red Blood Cell (million/cmm) were 14.1±1.4 and 5.1±0.4 respectively. Mean±SD of serum ferritin level (ng/ml) was 96.4±69.0ng/ml with a range of 4.1ng/ml to 298.7ng/ml. Among the respondents 9.0% had depleted iron store, 7.0 reduced iron store and 84.0% had normal iron store. Among the respondents 5.0% had iron deficiency anaemia in term of serum ferritin level. Statistically significant difference of serum ferritin level observed between male and female and donors with and without history of previous blood donation. Among the healthy blood donors of Bangladesh abnormal serum ferritin is highly prevalent among blood donors specially among female. Monitoring of iron stores by serum ferritin seems justified in order to identify those with depleted iron stores who will

  3. Gas diffusion liquid storage bag and method of use for storing blood

    NASA Technical Reports Server (NTRS)

    Bank, H.; Cleland, E. L. (Inventor)

    1979-01-01

    The shelf life of stored whole blood may be doubled by adding a buffer which maintains a desired pH level. However, this buffer causes the generation of CO2 which, if not removed at a controlled rate, causes the pH value of the blood to decrease, which shortens the useful life of the blood. A blood storage bag is described which permits the CO2 to be diffused out at a controlled rate into the atmosphere, thereby maintaining the desired pH value and providing a bag strong enough to permit handling.

  4. Effects of Aged Stored Autologous Red Blood Cells on Human Endothelial Function

    PubMed Central

    Kanias, Tamir; Triulzi, Darrel; Donadee, Chenell; Barge, Suchitra; Badlam, Jessica; Jain, Shilpa; Belanger, Andrea M.; Kim-Shapiro, Daniel B.

    2015-01-01

    Rationale: A major abnormality that characterizes the red cell “storage lesion” is increased hemolysis and reduced red cell lifespan after infusion. Low levels of intravascular hemolysis after transfusion of aged stored red cells disrupt nitric oxide (NO) bioavailabity, via accelerated NO scavenging reaction with cell-free plasma hemoglobin. The degree of intravascular hemolysis post-transfusion and effects on endothelial-dependent vasodilation responses to acetylcholine have not been fully characterized in humans. Objectives: To evaluate the effects of blood aged to the limits of Food and Drug Administration–approved storage time on the human microcirculation and endothelial function. Methods: Eighteen healthy individuals donated 1 U of leukopheresed red cells, divided and autologously transfused into the forearm brachial artery 5 and 42 days after blood donation. Blood samples were obtained from stored blood bag supernatants and the antecubital vein of the infusion arm. Forearm blood flow measurements were performed using strain-gauge plethysmography during transfusion, followed by testing of endothelium-dependent blood flow with increasing doses of intraarterial acetylcholine. Measurements and Main Results: We demonstrate that aged stored blood has higher levels of arginase-1 and cell-free plasma hemoglobin. Compared with 5-day blood, the transfusion of 42-day packed red cells decreases acetylcholine-dependent forearm blood flows. Intravascular venous levels of arginase-1 and cell-free plasma hemoglobin increase immediately after red cell transfusion, with more significant increases observed after infusion of 42-day-old blood. Conclusions: We demonstrate that the transfusion of blood at the limits of Food and Drug Administration–approved storage has a significant effect on the forearm circulation and impairs endothelial function. Clinical trial registered with www.clinicaltrials.gov (NCT 01137656) PMID:26222884

  5. Effects of Aged Stored Autologous Red Blood Cells on Human Endothelial Function.

    PubMed

    Risbano, Michael G; Kanias, Tamir; Triulzi, Darrel; Donadee, Chenell; Barge, Suchitra; Badlam, Jessica; Jain, Shilpa; Belanger, Andrea M; Kim-Shapiro, Daniel B; Gladwin, Mark T

    2015-11-15

    A major abnormality that characterizes the red cell "storage lesion" is increased hemolysis and reduced red cell lifespan after infusion. Low levels of intravascular hemolysis after transfusion of aged stored red cells disrupt nitric oxide (NO) bioavailabity, via accelerated NO scavenging reaction with cell-free plasma hemoglobin. The degree of intravascular hemolysis post-transfusion and effects on endothelial-dependent vasodilation responses to acetylcholine have not been fully characterized in humans. To evaluate the effects of blood aged to the limits of Food and Drug Administration-approved storage time on the human microcirculation and endothelial function. Eighteen healthy individuals donated 1 U of leukopheresed red cells, divided and autologously transfused into the forearm brachial artery 5 and 42 days after blood donation. Blood samples were obtained from stored blood bag supernatants and the antecubital vein of the infusion arm. Forearm blood flow measurements were performed using strain-gauge plethysmography during transfusion, followed by testing of endothelium-dependent blood flow with increasing doses of intraarterial acetylcholine. We demonstrate that aged stored blood has higher levels of arginase-1 and cell-free plasma hemoglobin. Compared with 5-day blood, the transfusion of 42-day packed red cells decreases acetylcholine-dependent forearm blood flows. Intravascular venous levels of arginase-1 and cell-free plasma hemoglobin increase immediately after red cell transfusion, with more significant increases observed after infusion of 42-day-old blood. We demonstrate that the transfusion of blood at the limits of Food and Drug Administration-approved storage has a significant effect on the forearm circulation and impairs endothelial function. Clinical trial registered with www.clinicaltrials.gov (NCT 01137656).

  6. Pediatric blood culture: time to positivity.

    PubMed

    Kara, Ateş; Kanra, Güler; Cengiz, A Bülent; Apiş, Menekşe; Gür, Deniz

    2004-01-01

    The aim of this study was to determine how long it takes blood culture to become positive using a blood culture system that can be monitored continuously in pediatric patients. Data were collected prospectively on 1,000 positive blood culture results from a tertiary pediatric university hospital from April 2000 to May 2002. The laboratory used the BACTEC 9120 fluorescent blood culture system. Patient's age ranged from less than a day to 20 years of age (mean 3 years). Five hundred and four cultures (50.4%) out of 1,000 yielded coagulase negative staphylococcus (CNS), 81 (8.1%) S. aureus, 53 (5.3%). Pseudomonas and 50 (5.0%) Klebsiella species. Of the 504 coagulase negative staphylococcal blood culture isolates, 314 (62.3% of CNS) were regarded as skin contaminants. Of the 1,000 cultures, 9.6% were reported as positive in the first day, 27.8% in the second day, 54.7% in the third day, 77.0% in the fourth and 89.4% in the fifth day. There was no association between previous antibiotic usage and the period required for isolate recovery. The clinician can expect to get results of positive blood cultures with susceptibility data, at a rate of 77.1% by day four and almost 90% by day five of sampling in the bacteriemic patient. Blood cultures yielding coagulase negative staphylococci in the first three days almost always show bacteremia with those microorganisms.

  7. Diving Related Changes in the Blood Oxygen Stores of Rehabilitating Harbor Seal Pups (Phoca vitulina)

    PubMed Central

    Thomas, Amber; Ono, Kathryn

    2015-01-01

    Harbor seal (Phoca vitulina) pups begin diving within hours of birth, stimulating the development of the blood oxygen (O2) stores necessary to sustain underwater aerobic metabolism. Since harbor seals experience a brief nursing period, the early-life development of these blood O2 stores is necessary for successful post-weaning foraging. If mothers and pups become prematurely separated, the pup may be transported to a wildlife rehabilitation center for care. Previous studies suggest that the shallow pools and lack of diving in rehabilitation facilities may lead to under-developed blood O2 stores, but diving behavior during rehabilitation has not been investigated. This study aimed to simultaneously study the diving behaviors and blood O2 store development of rehabilitating harbor seal pups. Standard hematology measurements (Hct, Hb, RBC, MCV, MCH, MCHC) were taken to investigate O2 storage capacity and pups were equipped with time-depth recorders to investigate natural diving behavior while in rehabilitation. Linear mixed models of the data indicate that all measured blood parameters changed with age; however, when compared to literature values for wild harbor seal pups, rehabilitating pups have smaller red blood cells (RBCs) that can store less hemoglobin (Hb) and subsequently, less O2, potentially limiting their diving capabilities. Wild pups completed longer dives at younger ages (maximum reported <25 days of age: 9 min) in previous studies than the captive pups in this study (maximum <25 days of age: 2.86 min). However, captivity may only affect the rate of development, as long duration dives were observed (maximum during rehabilitation: 13.6 min at 89 days of age). Further, this study suggests that there may be a positive relationship between RBC size and the frequency of long duration dives. Thus, rehabilitating harbor seal pups should be encouraged to make frequent, long duration dives to prepare themselves for post-release foraging. PMID:26061662

  8. Diving Related Changes in the Blood Oxygen Stores of Rehabilitating Harbor Seal Pups (Phoca vitulina).

    PubMed

    Thomas, Amber; Ono, Kathryn

    2015-01-01

    Harbor seal (Phoca vitulina) pups begin diving within hours of birth, stimulating the development of the blood oxygen (O2) stores necessary to sustain underwater aerobic metabolism. Since harbor seals experience a brief nursing period, the early-life development of these blood O2 stores is necessary for successful post-weaning foraging. If mothers and pups become prematurely separated, the pup may be transported to a wildlife rehabilitation center for care. Previous studies suggest that the shallow pools and lack of diving in rehabilitation facilities may lead to under-developed blood O2 stores, but diving behavior during rehabilitation has not been investigated. This study aimed to simultaneously study the diving behaviors and blood O2 store development of rehabilitating harbor seal pups. Standard hematology measurements (Hct, Hb, RBC, MCV, MCH, MCHC) were taken to investigate O2 storage capacity and pups were equipped with time-depth recorders to investigate natural diving behavior while in rehabilitation. Linear mixed models of the data indicate that all measured blood parameters changed with age; however, when compared to literature values for wild harbor seal pups, rehabilitating pups have smaller red blood cells (RBCs) that can store less hemoglobin (Hb) and subsequently, less O2, potentially limiting their diving capabilities. Wild pups completed longer dives at younger ages (maximum reported <25 days of age: 9 min) in previous studies than the captive pups in this study (maximum <25 days of age: 2.86 min). However, captivity may only affect the rate of development, as long duration dives were observed (maximum during rehabilitation: 13.6 min at 89 days of age). Further, this study suggests that there may be a positive relationship between RBC size and the frequency of long duration dives. Thus, rehabilitating harbor seal pups should be encouraged to make frequent, long duration dives to prepare themselves for post-release foraging.

  9. Washing stored red blood cells in an albumin solution improves their morphological and hemorheological properties

    PubMed Central

    Reinhart, Walter H.; Piety, Nathaniel Z.; Deuel, Jeremy W.; Makhro, Asya; Schulzki, Thomas; Bogdanov, Nikolay; Goede, Jeroen S.; Bogdanova, Anna; Abidi, Rajaa; Shevkoplyas, Sergey S.

    2015-01-01

    BACKGROUND Prolonged storage of red blood cells leads to storage lesions, which may impair clinical outcomes after transfusion. A hallmark of storage lesions is progressive echinocytic shape transformation, which can be partially reversed by washing in albumin solutions. Here we have investigated the impact of this shape recovery on biorheological parameters. METHODS Red blood cells stored hypothermically for 6–7 weeks were washed in a 1% human serum albumin solution. Red cell deformability was measured with osmotic gradient ektacytometry. The viscosity of red cell suspensions were measured with a Couette-type viscometer. The flow behaviour of red cells suspended at 40% hematocrit was tested with an artificial microvascular network. RESULTS Washing in 1% albumin reduced higher degrees of echinocytes and increased the frequency of discocytes, thereby shifting the morphological index towards discocytosis. Washing also reduced red cell swelling. This shape recovery was not seen after washing in saline, buffer or plasma. Red cell shape normalisation did not improve cell deformability measured by ektacytometry, but it tended to decrease suspension viscosities at low shear rates and improved the perfusion of an artificial microvascular network. CONCLUSIONS Washing of stored red blood cells in a 1% human serum albumin solution specifically reduces echinocytosis, and this shape recovery has a beneficial effect on microvascular perfusion in vitro. Washing in 1% albumin may represent a new approach to improving the quality of stored red cells, and thus potentially reducing the likelihood of adverse clinical outcomes associated with transfusion of blood stored for longer periods of time. PMID:25752902

  10. The possible advantages of cryoprecipitate prepared from fresh frozen plasma from blood stored for 24 hours.

    PubMed

    Philip, Joseph; Kumarage, Samantha; Chatterjee, Tathagata; Kumar, Sudeep; Mallhi, Rajeev

    2014-01-01

    To compare the coagulation-factor profile of cryoprecipitate produced from fresh frozen plasma from whole blood (WB) stored for 24 hours at room temperature (24CP) with that of standard cryoprecipitate (CP). We collected 80 units of WB from healthy volunteers, of which 20 units were of each blood group. Each unit of blood was divided into 2 parts. One part was used for preparation and quality-control evaluation of CP within 8 hours of collection; the other part was stored at room temperature for 24 hours and then subjected to CP preparation. Coagulation studies were carried out on each batch of CP after production. Fibrinogen, Factor VIII (FVII), and von Willibrand factor (vWF) were measured, and the blood groups were determined. We used the Student's t-test to perform comparisons and considered results to be significant at P < .005. Overall, all 3 clotting factors were increased in 24CP compared with CP, with a statistically significant increase in the level of FVIII. Blood group AB had significantly increased levels of fibrinogen and vWF in 24CP compared with CP. Our study showed that 24CP has equal or greater levels of coagulation factors compared with CP. His indicates that our alternate approach for preparation of CP may enable more efficient use of blood collected in satellite blood collection centers and during blood drives.

  11. Metabolic pathways that correlate with post-transfusion circulation of stored murine red blood cells

    PubMed Central

    de Wolski, Karen; Fu, Xiaoyoun; Dumont, Larry J.; Roback, John D.; Waterman, Hayley; Odem-Davis, Katherine; Howie, Heather L.; Zimring, James C.

    2016-01-01

    Transfusion of red blood cells is a very common inpatient procedure, with more than 1 in 70 people in the USA receiving a red blood cell transfusion annually. However, stored red blood cells are a non-uniform product, based upon donor-to-donor variation in red blood cell storage biology. While thousands of biological parameters change in red blood cells over storage, it has remained unclear which changes correlate with function of the red blood cells, as opposed to being co-incidental changes. In the current report, a murine model of red blood cell storage/transfusion is applied across 13 genetically distinct mouse strains and combined with high resolution metabolomics to identify metabolic changes that correlated with red blood cell circulation post storage. Oxidation in general, and peroxidation of lipids in particular, emerged as changes that correlated with extreme statistical significance, including generation of dicarboxylic acids and monohydroxy fatty acids. In addition, differences in anti-oxidant pathways known to regulate oxidative stress on lipid membranes were identified. Finally, metabolites were identified that differed at the time the blood was harvested, and predict how the red blood cells perform after storage, allowing the potential to screen donors at time of collection. Together, these findings map out a new landscape in understanding metabolic changes during red blood cell storage as they relate to red blood cell circulation. PMID:26921359

  12. Metabolic pathways that correlate with post-transfusion circulation of stored murine red blood cells.

    PubMed

    de Wolski, Karen; Fu, Xiaoyoun; Dumont, Larry J; Roback, John D; Waterman, Hayley; Odem-Davis, Katherine; Howie, Heather L; Zimring, James C

    2016-05-01

    Transfusion of red blood cells is a very common inpatient procedure, with more than 1 in 70 people in the USA receiving a red blood cell transfusion annually. However, stored red blood cells are a non-uniform product, based upon donor-to-donor variation in red blood cell storage biology. While thousands of biological parameters change in red blood cells over storage, it has remained unclear which changes correlate with function of the red blood cells, as opposed to being co-incidental changes. In the current report, a murine model of red blood cell storage/transfusion is applied across 13 genetically distinct mouse strains and combined with high resolution metabolomics to identify metabolic changes that correlated with red blood cell circulation post storage. Oxidation in general, and peroxidation of lipids in particular, emerged as changes that correlated with extreme statistical significance, including generation of dicarboxylic acids and monohydroxy fatty acids. In addition, differences in anti-oxidant pathways known to regulate oxidative stress on lipid membranes were identified. Finally, metabolites were identified that differed at the time the blood was harvested, and predict how the red blood cells perform after storage, allowing the potential to screen donors at time of collection. Together, these findings map out a new landscape in understanding metabolic changes during red blood cell storage as they relate to red blood cell circulation.

  13. Impact of non-storing biomass on PHA production: an enrichment culture on acetate and methanol.

    PubMed

    Marang, Leonie; Jiang, Yang; van Loosdrecht, Mark C M; Kleerebezem, Robbert

    2014-11-01

    The use of enrichment cultures for polyhydroxyalkanoate (PHA) production from substrate mixtures such as wastewater inevitably results in the establishment of a non-PHA-storing population besides the PHA-producing bacteria. This reduces the maximum PHA content that can be established, and increases downstream-processing costs. The aim of this study was to investigate the impact of non-storing biomass on the PHA production process. A microbial culture was enriched in a sequencing batch reactor fed with acetate and methanol. Methanol served as model substrate for compounds unsuitable for PHA production. The enrichment was dominated by Plasticicumulans acidivorans, a known PHA producer, and Methylobacillus flagellatus, an obligate methylotroph that cannot store PHA. As expected, the presence of the non-storing population lowered the maximum PHA content of the culture, from more than 80 to 66wt.%. To mimic a nitrogen-rich waste stream, additional accumulation experiments were performed with continuous supply of carbon and ammonium. In these experiments P. acidivorans still accumulated large amounts of PHA, but unrestricted growth of the non-storing, methylotrophic population reduced the maximum overall PHA content to 52wt.%. Besides ammonium limitation, other strategies to restrict the fraction of non-storing biomass should be developed. The mixture of acetate and methanol is a useful model substrate for the development of such strategies. Copyright © 2014 Elsevier B.V. All rights reserved.

  14. Analysis of RBC-microparticles in stored whole blood bags - a promising marker to detect blood doping in sports?

    PubMed

    Voss, Sven Christian; Jaganjac, Morana; Al-Thani, Amna Mohamed; Grivel, Jean-Charles; Raynaud, Christophe Michel; Al-Jaber, Hind; Al-Menhali, Afnan Saleh; Merenkov, Zeyed Ahmad; Alsayrafi, Mohammed; Latiff, Aishah; Georgakopoulos, Costas

    2017-05-04

    Blood doping in sports is prohibited by the World Anti-Doping Agency (WADA). To find a possible biomarker for the detection of blood doping, we investigated the changes in blood stored in CPDA-1 blood bags of eight healthy subjects who donated one unit of blood. Aliquots were taken on days 0, 14, and 35. Platelet-free plasma was prepared and stored at -80°C until analysis on a flow cytometer dedicated for the analysis of microparticles (MPs). Changes in the number of red blood cell (RBC) -MPs were highly significant (p < 0.0001) with a mean of 219 (10^3/μL) on day 0 changing to 23 120 (10^3/μL) on day 14 and 29 310 (10^3/μL) on day 35. We conclude that RBC-MPs seem to be a promising biomarker for doping control but confirmation by a transfusion study is necessary. Copyright © 2017 John Wiley & Sons, Ltd.

  15. Microfluidic evaluation of red cells collected and stored in modified processing solutions used in blood banking.

    PubMed

    Wang, Yimeng; Giebink, Adam; Spence, Dana M

    2014-01-01

    The most recent American Association of Blood Banks survey found that 40,000 units of blood are required daily for general medicine, hematology/oncology, surgery, and for accident and trauma victims. While blood transfusions are an extremely important component of critical healthcare, complications associated with transfusion of blood components still exist. It is well-established that the red blood cell (RBC) undergoes many physical and chemical changes during storage. Increased oxidative stress, formation of advanced glycation endproducts, and microparticle formation are all known to occur during RBC storage. Furthermore, it is also known that patients who receive a transfusion have reduced levels of available nitric oxide (NO), a major determinant in blood flow. However, the origin of this reduced NO bioavailability is not completely understood. Here, we show that a simple modification to the glucose concentration in the solutions used to process whole blood for subsequent RBC storage results in a remarkable change in the ability of these cells to stimulate NO. In a controlled in vitro microflow system, we discovered that storage of RBCs in normoglycemic versions of standard storage solutions resulted in RBC-derived ATP release values 4 weeks into storage that were significantly greater than day 1 release values for those RBCs stored in conventional solutions. During the same storage duration, microfluidic technologies enabled measurements of endothelium-derived NO that were stimulated by the ATP release from the stored RBCs. In comparison to currently accepted processing solutions, the NO production increased by more than 25% in the presence of the RBCs stored in the normoglycemic storage solutions. Control experiments using inhibitors of ATP release from the RBCs, or ATP binding to the endothelium, strongly suggest that the increased NO production by the endothelium is directly related to the ability of the stored RBCs to release ATP. We anticipate these

  16. Carbon nanotubes activate store-operated calcium entry in human blood platelets.

    PubMed

    Lacerda, Silvia H De Paoli; Semberova, Jana; Holada, Karel; Simakova, Olga; Hudson, Steven D; Simak, Jan

    2011-07-26

    Carbon nanotubes (CNTs) are known to potentiate arterial thrombosis in animal models, which raises serious safety issues concerning environmental or occupational exposure to CNTs and their use in various biomedical applications. We have shown previously that different CNTs, but not fullerene (nC60), induce the aggregation of human blood platelets. To date, however, a mechanism of potentially thrombogenic CNT-induced platelet activation has not been elucidated. Here we show that pristine multiwalled CNTs (MWCNTs) penetrate platelet plasma membrane without any discernible damage but interact with the dense tubular system (DTS) causing depletion of platelet intracellular Ca(2+) stores. This process is accompanied by the clustering of stromal interaction molecule 1 (STIM1) colocalized with Orai1, indicating the activation of store-operated Ca(2+) entry (SOCE). Our findings reveal the molecular mechanism of CNT-induced platelet activation which is critical in the evaluation of the biocompatibility of carbon nanomaterials with blood.

  17. Update on blood culture-negative endocarditis.

    PubMed

    Tattevin, P; Watt, G; Revest, M; Arvieux, C; Fournier, P-E

    2015-01-01

    Blood culture-negative endocarditis is often severe, and difficult to diagnose. The rate of non-documented infective endocarditis has decreased with the advent of molecular biology - improved performance for the diagnosis of bacterial endocarditis with blood cultures sterilized by previous antibacterial treatment - and cardiac surgery - access to the main infected focus, the endocardium, for half of the patients. Blood culture-negative endocarditis are classified in 3 main categories: (i) bacterial endocarditis with blood cultures sterilized by previous antibacterial treatment (usually due to usual endocarditis-causing bacteria, i.e. streptococci, more rarely staphylococci, or enterococci); (ii) endocarditis related to fastidious microorganisms (e.g. HACEK bacteria; defective streptococci - Gemella, Granulicatella, and Abiotrophia sp. - Propionibacterium acnes, Candida sp.): in these cases, prolonged incubation will allow identifying the causative pathogen in a few days; (iii) and the "true" blood culture-negative endocarditis, due to intra-cellular bacteria that cannot be routinely cultured in blood with currently available techniques: in France, these are most frequently Bartonella sp., Coxiella burnetti (both easily diagnosed by ad hoc serological tests), and Tropheryma whipplei (usually diagnosed by PCR on excised cardiac valve tissue). Non-infective endocarditis is rare, mostly limited to marantic endocarditis, and the rare endocarditis related to systemic diseases (lupus, Behçet).

  18. Platelets in blood stored in untreated and siliconed glass bottles and plastic bags. II. Survival studies.

    PubMed

    KISSMEYER-NIELSEN, F; MADSEN, C B

    1961-11-01

    Platelet survival was determined using untreated and siliconed glass bottles and plastic bags (Fenwal) for collecting and storing blood. The platelets were tagged in vivo with P(32) in six polycythaemic patients undergoing treatment with P(32). The results showed that fresh ACD blood collected in untreated glass, siliconed glass, and plastic gave the same recovery of platelets in the recipients. The use of EDTA (Fenwal formula) as anticoagulant gave results inferior to those obtained with blood using ACD as anticoagulant. Even after storage up to 24 hours in untreated glass bottles (ordinary bank blood) a satisfactory recovery of platelets was observed. After storage for 72 hours the recovery was less but not negligible.

  19. Laboratory contamination of blood cultures.

    PubMed

    Spencer, R C; Savage, M A

    1975-12-01

    A prospective study of the use of a laminar flow cabinet, an exhaust-ventilated safety hood, and the open bench for the microbiological examination of blood is described. Blood samples from 1600 patients were subcultured on the open bench, 2700 in a safety hood, and 2607 in a laminar flow cabinet. Use of the laminar flow cabinet produced a significantly greater level of contamination than the other methods, and it is concluded that the exhaust-ventilated safety hood should be used for this procedure.

  20. Reduction in potassium concentration of stored red blood cell units using a resin filter.

    PubMed

    Yamada, Chisa; Heitmiller, Eugenie S; Ness, Paul M; King, Karen E

    2010-09-01

    Hyperkalemia is a serious complication of rapid and massive blood transfusion due to high plasma potassium (K) in stored red blood cell (RBC) units. A potassium adsorption filter (PAF) was developed in Japan to remove K by exchanging with sodium (Na). We performed an in vitro evaluation of its efficacy and feasibility of use. Three AS-3 RBC units were filtered by each PAF using gravity; 10 PAFs were tested. Blood group, age, flow rate, and irradiation status were recorded. Total volume, K, Na, Cl, Mg, total Ca (tCa), RBC count, hemoglobin (Hb), hematocrit (Hct), and plasma Hb were measured before and after filtering each unit. Ionized Ca (iCa), pH, and glucose were measured for some units. After filtration, the mean decrease in K was 97.5% in the first RBC unit, 91.2% in the second unit, and 64.4% in the third unit. The mean increases in Na, Mg, and tCa were 33.0, 151.4, and 116.1%, respectively. iCa and pH remained low; glucose was unchanged. RBC count, Hb, and Hct decreased slightly after filtration of first units; plasma Hb was unchanged. After filtration, there was no visual evidence of increased hemolysis or clot formation. The PAF decreased K concentration in stored AS-3 RBC units to minimal levels in the first and second RBC units. Optimally, one filter could be used for 2 RBC units. Although Na increased, the level may not be clinically significant. PAF may be useful for at-risk patients receiving older units or blood that has been stored after gamma irradiation. © 2010 American Association of Blood Banks.

  1. Clinical implications of positive blood cultures.

    PubMed Central

    Bryan, C S

    1989-01-01

    Positive blood cultures can be classified according to their veracity (true-positive or false-positive culture), clinical severity (inconsequential or life threatening), place of origin (community acquired or nosocomial), source (primary or secondary), duration (transient, intermittent, or continuous), pattern of occurrence (single episode, persistent, or recurrent), or intensity (high or low grade). In general, however, positive blood cultures identify a patient population at high risk of death. In my studies, patients with positive blood cultures were 12 times more likely to die during hospitalization than patients without positive blood cultures. Many bacteremias and fungemias occur in complicated clinical settings, and it appears that only about one-half of the deaths among affected patients are due directly to infection. Hence, it is appropriate to speak of "crude mortality" and "attributable mortality." Among hospitalized patients, recent trends include rising incidences of Staphylococcus aureus and coagulase-negative staphylococcal and enterococcal bacteremias and a dramatic increase in the incidence of fungemias. The diagnostic and therapeutic implications of blood cultures positive for specific microorganisms continue to evolve and are the subject of a large and growing medical literature. PMID:2680055

  2. Effect of Iron Supplementation on Iron Stores and Total Body Iron after Whole Blood Donation

    PubMed Central

    Cable, Ritchard G.; Brambilla, Donald; Glynn, Simone A.; Kleinman, Steven; Mast, Alan E.; Spencer, Bryan R.; Stone, Mars; Kiss, Joseph E.

    2016-01-01

    Background Understanding the effect of blood donation and iron supplementation on iron balance will inform strategies to manage donor iron status Study Design and Methods 215 donors were randomized to receive ferrous gluconate daily (37.5 mg iron) or no iron for 24 weeks after blood donation. Iron stores were assessed using ferritin and soluble Transferrin Receptor. Hemoglobin iron was calculated from total body hemoglobin. Total Body Iron (TBI) was estimated by summing iron stores and hemoglobin iron. Results At 24 weeks, TBI in donors taking iron increased by 281.0 mg (95% Confidence Interval [CI]: 223.4, 338.6) compared to pre-donation, while TBI in donors not on iron decreased by 74.1 mg (CI: −112.3, −35.9), p<0.0001, iron vs. no iron. TBI increased rapidly following blood donation with iron supplementation, especially in iron depleted donors. Supplementation increased TBI compared to controls during the first 8 weeks after donation: 367.8 mg (CI: 293.5, 442.1) versus −24.1 mg (CI: −82.5, 34.3) for donors with baseline ferritin ≤26 ng/mL; and 167.8 mg (95%CI: 116.5, 219.2) versus −68.1 mg (CI: −136.7, 0.5) for donors with baseline ferritin >26 ng/mL. 88% of the benefit of iron supplementation occurred during the first 8 weeks after blood donation. Conclusion Donors on iron supplementation replaced donated iron while donors not on iron did not. Eight weeks of iron supplementation provided nearly all of the measured improvement in TBI. Daily iron supplementation after blood donation allows blood donors to recover the iron loss from blood donation and prevents sustained iron deficiency. PMID:27232535

  3. Effect of iron supplementation on iron stores and total body iron after whole blood donation.

    PubMed

    Cable, Ritchard G; Brambilla, Donald; Glynn, Simone A; Kleinman, Steven; Mast, Alan E; Spencer, Bryan R; Stone, Mars; Kiss, Joseph E

    2016-08-01

    Understanding the effect of blood donation and iron supplementation on iron balance will inform strategies to manage donor iron status. A total of 215 donors were randomized to receive ferrous gluconate daily (37.5 mg iron) or no iron for 24 weeks after blood donation. Iron stores were assessed using ferritin and soluble transferrin receptor. Hemoglobin (Hb) iron was calculated from total body Hb. Total body iron (TBI) was estimated by summing iron stores and Hb iron. At 24 weeks, TBI in donors taking iron increased by 281.0 mg (95% confidence interval [CI], 223.4-338.6 mg) compared to before donation, while TBI in donors not on iron decreased by 74.1 mg (95% CI, -112.3 to -35.9; p < 0.0001, iron vs. no iron). TBI increased rapidly after blood donation with iron supplementation, especially in iron-depleted donors. Supplementation increased TBI compared to controls during the first 8 weeks after donation: 367.8 mg (95% CI, 293.5-442.1) versus -24.1 mg (95% CI, -82.5 to 34.3) for donors with a baseline ferritin level of not more than 26 ng/mL and 167.8 mg (95% CI, 116.5-219.2) versus -68.1 mg (95% CI, -136.7 to 0.5) for donors with a baseline ferritin level of more than 26 ng/mL. A total of 88% of the benefit of iron supplementation occurred during the first 8 weeks after blood donation. Donors on iron supplementation replaced donated iron while donors not on iron did not. Eight weeks of iron supplementation provided nearly all of the measured improvement in TBI. Daily iron supplementation after blood donation allows blood donors to recover the iron loss from blood donation and prevents sustained iron deficiency. © 2016 AABB.

  4. Time Dependent Assessment of Morphological Changes: Leukodepleted Packed Red Blood Cells Stored in SAGM.

    PubMed

    Mustafa, Ibrahim; Al Marwani, Asma; Mamdouh Nasr, Khuloud; Abdulla Kano, Noora; Hadwan, Tameem

    2016-01-01

    Usually packed red blood cells (pRBCs) require specific conditions in storage procedures to ensure the maximum shelf life of up to 42 days in 2-6°C. However, molecular and biochemical consequences can affect the stored blood cells; these changes are collectively labeled as storage lesions. In this study, the effect of prolonged storage was assessed through investigating morphological changes and evaluating oxidative stress. Samples from leukodepleted pRBC in SAGM stored at 4°C for 42 days were withdrawn aseptically on day 0, day 14, day 28, and day 42. Morphological changes were observed using scanning electron microscopy and correlated with osmotic fragility and hematocrit. Oxidative injury was studied through assessing MDA level as a marker for lipid peroxidation. Osmotic fragility test showed that extended storage time caused increase in the osmotic fragility. The hematocrit increased by 6.6% from day 0 to day 42. The last 2 weeks show alteration in the morphology with the appearance of echinocytes and spherocytes. Storage lesions and morphological alterations appeared to affect RBCs during the storage period. Further studies should be performed to develop strategies that will aid in the improvement of stored pRBC quality and efficacy.

  5. Insulin sensitivity, vascular function, and iron stores in voluntary blood donors.

    PubMed

    Zheng, Haoyi; Patel, Milan; Cable, Ritchard; Young, Lawrence; Katz, Stuart D

    2007-10-01

    Reduced iron stores after blood donation are associated with improved vascular function and decreased cardiovascular risk. We sought to determine whether iron-dependent changes in glucose metabolism may contribute to improved vascular function in blood donors. We conducted a prospective cross-sectional study in 21 high-frequency blood donors (more than eight donations in the last 2 years) and 21 low-frequency blood donors (one to two donations in the last 2 years) aged 50-75 years. Serum markers of iron stores, whole-body insulin sensitivity index (WBISI) during oral glucose tolerance testing, and flow-mediated dilation in the brachial artery were determined in all subjects. Serum ferritin was decreased (median values 23 vs. 36 ng/ml, P < 0.05) and flow-mediated dilation in the brachial artery was increased (median values 5.9 vs. 5.3%, P < 0.05) in high-frequency donors compared with low-frequency donors, respectively, but WBISI (median values 4.8 vs. 4.7) and related measures of glucose tolerance did not differ between groups. Flow-mediated dilation significantly decreased at 1 h after oral glucose loading in both groups, but the decrease in flow-mediated dilation at 1 h did not differ between high- and low-frequency donors. High-frequency blood donation reduced serum ferritin and increased flow-mediated dilation compared with low-frequency donation but did not improve insulin sensitivity or protect the vascular endothelium from the adverse effects of acute hyperglycemia after oral glucose loading. These findings suggest that the mechanisms linking blood donation to improved vascular function are not likely related to changes in glucose metabolism.

  6. Blood iron stores reduction affects lipoprotein status--a potential benefit of blood donation.

    PubMed

    Jadrić, Radivoj; Hasić, Sabaheta; Kiseljaković, Emina; Corić, Jozo; Prnjavorac, Besim; Winterhalter-Jadrić, Mira

    2011-02-01

    To determine the lipoprotein profile of voluntary blood donors, and on the basis of parameters to evaluate the risk of atherosclerosis. The study included voluntary blood donors of both sexes. Participants were divided into two groups. The first group of subjects consisted of men and women in menopause (BD 1). The second group consisted of women in reproductive age (BD 2). Analysis of concentration of lipoproteins was performed by direct determination of total cholesterol, LDL-C and HDL-C. From the total serum cholesterol and concentration of lipoproteins ratios of total cholesterol/HDL-C ratio and LDL-C/HDL-C were calculated. Significantly higher concentration of LDL-C was obtained in the serum of BD 1, compared to LDL-C in the serum of BD 2, within the reference range. Mean concentration of HDL-C in the serum of BD 2 group was higher than the values measured in the BD group 1, without significant difference. The ratio of total cholesterol/HDL-C showed significantly higher values in the BD 1 group compared with results in the BD 2 group. Significantly higher values in the BD group 1 were observed for the ratio of LDL-C/HDL-C. Obtained results showed that all voluntary blood donors had a concentration of individual lipoprotein fractions in a lower risk range for atherosclerosis development. Female voluntary blood donors in reproductive age have a more favorable lipid status in relation to the voluntary blood donors, men and women in menopause, indicating that this population of women is exposed to lower risk of developing atherosclerosis.

  7. Prospects of Vitamin C as an Additive in Plasma of Stored Blood

    PubMed Central

    Vani, R.; Soumya, R.; Carl, H.; Chandni, V. A.; Neha, K.; Pankhuri, B.; Trishna, S.; Vatsal, D. P.

    2015-01-01

    There is a dire necessity to improve blood storage and prolong shelf-life of blood. Very few studies have focused on oxidative stress (OS) in blood and its influence on plasma with storage. This study attempts to (i) elucidate the continuous changes occurring in plasma during storage through oxidant levels and antioxidant status and (ii) evaluate the influence of vitamin C (VC) as an additive during blood storage. Blood was drawn from male Wistar rats and stored for 25 days at 4°C. Blood samples were divided into control and experimental groups. Plasma was isolated every 5 days and the OS markers, antioxidant enzymes, lipid peroxidation, and protein oxidation products, were studied. Catalase activity increased in all groups with storage. Lipid peroxidation decreased in VC (10) but was maintained in VC (30) and VC (60). Although there were variations in all groups, carbonyls were maintained towards the end of storage. Advanced oxidation protein products (AOPP) increased in VC (30) and were maintained in VC (10) and VC (60). Sulfhydryls were maintained in all groups. Vitamin C could not sufficiently attenuate OS and hence, this opens the possibilities for further studies on vitamin C in combination with other antioxidants, in storage solutions. PMID:26345502

  8. The quality assessment of stored red blood cells probed using atomic-force microscopy.

    PubMed

    Lamzin, I M; Khayrullin, R M

    2014-01-01

    At the moment the suitability of stored red blood cells (sRBC) for transfusion is checked by routine methods such as haemoglobin estimation and the level of haemolysis. These methods cannot characterize directly the quality of the membranes of sRBC. The aim of this work is to assess the quality of sRBC based on such criteria as the membrane's stiffness and the size and the form of sRBC. Materials and Methods. We have investigated 5 series of dry cytosmears of the sRBC which had been kept in blood bank in a period from 1 to 35 days. After AFM imaging, in every specimen, 5 RBC were chosen at random; the diameter, the height, and the stiffness were measured on each of them. Results. The present study shows high increase of the mean values of YM and height of RBC after 35 days of storage and decrease of the mean values of their diameter. Conclusion. Statistically significant high increase of the mean values of YM indicates the decrease of the elasticity of the cells in the course of storing of the RBC. This parameter along with the morphological characteristics can be used as criterion for assessment of applicability of the sRBC for blood transfusion.

  9. The Quality Assessment of Stored Red Blood Cells Probed Using Atomic-Force Microscopy

    PubMed Central

    Lamzin, I. M.; Khayrullin, R. M.

    2014-01-01

    At the moment the suitability of stored red blood cells (sRBC) for transfusion is checked by routine methods such as haemoglobin estimation and the level of haemolysis. These methods cannot characterize directly the quality of the membranes of sRBC. The aim of this work is to assess the quality of sRBC based on such criteria as the membrane's stiffness and the size and the form of sRBC. Materials and Methods. We have investigated 5 series of dry cytosmears of the sRBC which had been kept in blood bank in a period from 1 to 35 days. After AFM imaging, in every specimen, 5 RBC were chosen at random; the diameter, the height, and the stiffness were measured on each of them. Results. The present study shows high increase of the mean values of YM and height of RBC after 35 days of storage and decrease of the mean values of their diameter. Conclusion. Statistically significant high increase of the mean values of YM indicates the decrease of the elasticity of the cells in the course of storing of the RBC. This parameter along with the morphological characteristics can be used as criterion for assessment of applicability of the sRBC for blood transfusion. PMID:25610651

  10. Evaluation of Verigene Blood Culture Test Systems for Rapid Identification of Positive Blood Cultures.

    PubMed

    Kim, Jae-Seok; Kang, Go-Eun; Kim, Han-Sung; Kim, Hyun Soo; Song, Wonkeun; Lee, Kyu Man

    2016-01-01

    The performance of molecular tests using the Verigene Gram-Positive and Gram-Negative Blood Culture nucleic acid tests (BC-GP and BC-GN, resp.; Naosphere, Northbrook, IL, USA) was evaluated for the identification of microorganisms detected from blood cultures. Ninety-nine blood cultures containing Gram-positive bacteria and 150 containing Gram-negative bacteria were analyzed using the BC-GP and BC-GN assays, respectively. Blood cultures were performed using the Bactec blood culture system (BD Diagnostic Systems, Franklin Lakes, NJ, USA) and conventional identification and antibiotic-susceptibility tests were performed using a MicroScan system (Siemens, West Sacramento, CA, USA). When a single strain of bacteria was isolated from the blood culture, Verigene assays correctly identified 97.9% (94/96) of Gram-positive bacteria and 93.8% (137/146) of Gram-negative bacteria. Resistance genes mecA and vanA were correctly detected by the BC-GP assay, while the extended-spectrum β-lactamase CTX-M and the carbapenemase OXA resistance gene were detected from 30 cases cultures by the BC-GN assay. The BC-GP and BC-GN assays showed high agreement with conventional identification and susceptibility tests. These tests are useful for rapid identification of microorganisms and the detection of clinically important resistance genes from positive Bactec blood cultures.

  11. Comparison of stored red blood cell washing techniques for priming extracorporeal circuits.

    PubMed

    Sasaki, Jun; Tirotta, Christopher; Lim, Hyunsoo; Kubes, Kathleen; Salvaggio, Jane; Hannan, Robert; Burke, Redmond; Ojito, Jorge

    2017-09-01

    The aim of this study was to compare three different blood washing techniques and describe the differences for the composition of the washed red blood cells (RBC). Stored RBCs less than 5 days old were washed using three different techniques. 1) Washing with normal saline with the COBE Model 2991 blood processor in the blood bank (BB-S). 2) Washing with normal saline with the Continuous AutoTransfusion System (C.A.T.S) in the operating room (OR-S). 3) Washing with Plasma-Lyte with the C.A.T.S in the operating room (OR-PL). Then, we compared the values for hemoglobin (Hb), hematocrit (Hct), blood volume, RBC volume, lactate, glucose, sodium and potassium of the three different groups. Forty-five units of RBCs were washed and analyzed (15 for each technique). The OR-S RBCs, when compared to the BB-S RBCs, had lower hemoglobin (g/dL) (22.8 vs 24.1, p=0.006), lower hematocrit (%) (67 vs 71, p=0.006), higher RBC volume (ml) (161 vs 130, p<0.001), higher glucose (mg/dL) (185 vs 46, p<0.001) and lower sodium (mmol/L) (153 vs 158, p<0.001). When compared to the OR-S RBCs, the OR-PL RBCs showed higher potassium (mmol/L) (5.3 vs 2, p<0.001) and lower sodium (mmol/L) (129 vs 153, p<0.001). RBCs washed with an autotransfusion device had a higher RBC volume and more physiological levels of glucose and sodium when compared with the blood processor in the blood bank. It can be an alternative option to use RBCs washed with an autotransfusion device for priming the extracorporeal circuits utilized in patients undergoing cardiac surgery.

  12. Characterization of bacteriophages infecting clinical isolates of Pseudomonas aeruginosa stored in a culture collection

    PubMed Central

    Zanetti, C.C.S.; Mingrone, R.C.C.; Kisielius, J.J.; Ueda-Ito, M.; Pignatari, A.C.C.

    2013-01-01

    Some clinical isolates of Pseudomonas aeruginosa stored in our culture collection did not grow or grew poorly and showed lysis on the culture plates when removed from the collection and inoculated on MacConkey agar. One hypothesis was that bacteriophages had infected and killed those clinical isolates. To check the best storage conditions to maintain viable P. aeruginosa for a longer time, clinical isolates were stored at various temperatures and were grown monthly. We investigated the presence of phage in 10 clinical isolates of P. aeruginosa stored in our culture collection. Four strains of P. aeruginosa were infected by phages that were characterized by electron microscopy and isolated to assess their ability to infect. The best condition to maintain the viability of the strains during storage was in water at room temperature. Three Siphoviridae and two Myoviridae phages were visualized and characterized by morphology. We confirmed the presence of bacteriophages infecting clinical isolates, and their ability to infect and lyse alternative hosts. Strain PAO1, however, did not show lysis to any phage. Mucoid and multidrug resistant strains of P. aeruginosa showed lysis to 50% of the phages tested. PMID:23969975

  13. Performance of Gram staining on blood cultures flagged negative by an automated blood culture system.

    PubMed

    Peretz, A; Isakovich, N; Pastukh, N; Koifman, A; Glyatman, T; Brodsky, D

    2015-08-01

    Blood is one of the most important specimens sent to a microbiology laboratory for culture. Most blood cultures are incubated for 5-7 days, except in cases where there is a suspicion of infection caused by microorganisms that proliferate slowly, or infections expressed by a small number of bacteria in the bloodstream. Therefore, at the end of incubation, misidentification of positive cultures and false-negative results are a real possibility. The aim of this work was to perform a confirmation by Gram staining of the lack of any microorganisms in blood cultures that were identified as negative by the BACTEC™ FX system at the end of incubation. All bottles defined as negative by the BACTEC FX system were Gram-stained using an automatic device and inoculated on solid growth media. In our work, 15 cultures that were defined as negative by the BACTEC FX system at the end of the incubation were found to contain microorganisms when Gram-stained. The main characteristic of most bacteria and fungi growing in the culture bottles that were defined as negative was slow growth. This finding raises a problematic issue concerning the need to perform Gram staining of all blood cultures, which could overload the routine laboratory work, especially laboratories serving large medical centers and receiving a large number of blood cultures.

  14. Hepcidin is a Better Predictor of Iron Stores in Premenopausal Women than Blood Loss or Dietary Intake.

    PubMed

    Lim, Karen H C; Booth, Alison O; Nowson, Caryl A; Szymlek-Gay, Ewa A; Irving, David O; Riddell, Lynn J

    2016-09-02

    The relationship between dietary intake, circulating hepcidin and iron status in free-living premenopausal women has not been explored. This cross-sectional study aimed to identify dietary determinants of iron stores after accounting for blood loss and to determine whether iron intake predicts iron stores independently of hepcidin in a sample of Australian women. Three hundred thirty eight women aged 18-50 years were recruited. Total intake and food sources of iron were determined via food frequency questionnaire; the magnitude of menstrual losses was estimated by self-report; and blood donation volume was quantified using blood donation records and self-reported donation frequency. Serum samples were analysed for ferritin, hepcidin and C-reactive protein concentrations. Linear regression was used to investigate associations. Accounting for blood loss, each 1 mg/day increase in dietary iron was associated with a 3% increase in iron stores (p = 0.027); this association was not independent of hepcidin. Hepcidin was a more influential determinant of iron stores than blood loss and dietary factors combined (R² of model including hepcidin = 0.65; R² of model excluding hepcidin = 0.17, p for difference <0.001), and increased hepcidin diminished the positive association between iron intake and iron stores. Despite not being the biggest contributor to dietary iron intake, unprocessed meat was positively associated with iron stores, and each 10% increase in consumption was associated with a 1% increase in iron stores (p = 0.006). No other dietary factors were associated with iron stores. Interventions that reduce hepcidin production combined with dietary strategies to increase iron intake may be important means of improving iron status in women with depleted iron stores.

  15. Hepcidin is a Better Predictor of Iron Stores in Premenopausal Women than Blood Loss or Dietary Intake

    PubMed Central

    Lim, Karen H. C.; Booth, Alison O.; Nowson, Caryl A.; Szymlek-Gay, Ewa A.; Irving, David O.; Riddell, Lynn J.

    2016-01-01

    The relationship between dietary intake, circulating hepcidin and iron status in free-living premenopausal women has not been explored. This cross-sectional study aimed to identify dietary determinants of iron stores after accounting for blood loss and to determine whether iron intake predicts iron stores independently of hepcidin in a sample of Australian women. Three hundred thirty eight women aged 18–50 years were recruited. Total intake and food sources of iron were determined via food frequency questionnaire; the magnitude of menstrual losses was estimated by self-report; and blood donation volume was quantified using blood donation records and self-reported donation frequency. Serum samples were analysed for ferritin, hepcidin and C-reactive protein concentrations. Linear regression was used to investigate associations. Accounting for blood loss, each 1 mg/day increase in dietary iron was associated with a 3% increase in iron stores (p = 0.027); this association was not independent of hepcidin. Hepcidin was a more influential determinant of iron stores than blood loss and dietary factors combined (R2 of model including hepcidin = 0.65; R2 of model excluding hepcidin = 0.17, p for difference <0.001), and increased hepcidin diminished the positive association between iron intake and iron stores. Despite not being the biggest contributor to dietary iron intake, unprocessed meat was positively associated with iron stores, and each 10% increase in consumption was associated with a 1% increase in iron stores (p = 0.006). No other dietary factors were associated with iron stores. Interventions that reduce hepcidin production combined with dietary strategies to increase iron intake may be important means of improving iron status in women with depleted iron stores. PMID:27598194

  16. Quantifying morphological heterogeneity: a study of more than 1 000 000 individual stored red blood cells.

    PubMed

    Piety, N Z; Gifford, S C; Yang, X; Shevkoplyas, S S

    2015-10-01

    The morphology of red blood cells (RBCs) deteriorates progressively during hypothermic storage. The degree of deterioration varies between individual cells, resulting in a highly heterogeneous population of cells contained within each RBC unit. Current techniques capable of categorizing the morphology of individual stored RBCs are manual, laborious and error-prone procedures that limit the number of cells that can be studied. Our objective was to create a simple, automated system for high-throughput RBC morphology classification. A simple microfluidic device, designed to enable rapid, consistent acquisition of images of optimally oriented RBCs, was fabricated using soft lithography. A custom image analysis algorithm was developed to categorize the morphology of each individual RBC in the acquired images. The system was used to determine morphology of individual RBCs in several RBC units stored hypothermically for 6-8 weeks. The system was used to automatically determine the distribution of cell diameter within each morphological class for >1 000 000 individual stored RBCs (speed: >10 000 cells/h; accuracy: 91·9% low resolution, 75·3% high resolution). Diameter mean and standard deviation by morphology class were as follows: discocyte 7·80 ± 0·49 μm, echinocyte 1 7·61 ± 0·63 μm, echinocyte 2 7·02 ± 0·61 μm, echinocyte 3 6·47 ± 0·42 μm, sphero-echinocyte 6·01 ± 0·26 μm, spherocyte 6·02 ± 0·27 μm, stomatocyte 1 6·95 ± 0·61 μm and stomatocyte 2 7·32 ± 0·47 μm. The automated morphology classification procedure described in this study is significantly simpler, faster and less subjective than conventional manual procedures. The ability to evaluate the morphology of individual RBCs automatically, rapidly and in statistically significant numbers enabled us to perform the most extensive study of stored RBC morphology to date. © 2015 International Society of Blood Transfusion.

  17. Fibrin glue from stored human plasma. An inexpensive and efficient method for local blood bank preparation.

    PubMed

    Spotnitz, W D; Mintz, P D; Avery, N; Bithell, T C; Kaul, S; Nolan, S P

    1987-08-01

    European surgeons have used fibrin glue extensively during thoracic, cardiovascular, and general surgical operations. Until now, however, it has been available only as a commercial preparation made from pooled human plasma, and it has not been approved by the U.S. Food and Drug Administration for use in the United States because of a high associated risk of hepatitis and acquired immune deficiency syndrome. Methods of obtaining fibrinogen, an essential component of fibrin glue, from cryoprecipitate or fresh frozen plasma have been published recently. However, the cryoprecipitate method results in relatively low concentrations of fibrinogen, which can reduce glue effectiveness. The fresh frozen plasma method is more expensive and does not meet the standards of the American Association of Blood Banks for the "closed" system required for safe handling and management of blood component products. Both the cryoprecipitate and the fresh frozen plasma methods result in waste of unstable clotting factors. These factors are necessary to replace human plasma clotting deficiencies but are not necessary for the production of fibrin glue. The authors have developed an efficient, high-concentration blood bank method for producing and maintaining a local supply of a safer and less expensive but equally effective material derived from stored human plasma. This material is produced using approved blood bank techniques for a "closed" system in blood component production, thus reducing the risks of contamination and infection, and its fibrinogen concentration is higher than that of standard cryoprecipitate. The cost of 1 unit of this fibrin glue is comparable to that for 1 unit of cryoprecipitate and less than that for 1 unit of fresh frozen plasma.(ABSTRACT TRUNCATED AT 250 WORDS)

  18. N-acetylcysteine improves the quality of red blood cells stored for transfusion.

    PubMed

    Amen, Florencia; Machin, Andrea; Touriño, Cristina; Rodríguez, Ismael; Denicola, Ana; Thomson, Leonor

    2017-04-06

    Storage inflicts a series of changes on red blood cells (RBC) that compromise the cell survival and functionality; largely these alterations (storage lesions) are due to oxidative modifications. The possibility of improving the quality of packed RBC stored for transfusion including N-acetylcysteine (NAC) in the preservation solution was explored. Relatively high concentrations of NAC (20-25 mM) were necessary to prevent the progressive leakage of hemoglobin, while lower concentrations (≥2.5 mM) were enough to prevent the loss of reduced glutathione during the first 21 days of storage. Peroxiredoxin-2 was also affected during storage, with a progressive accumulation of disulfide-linked dimers and hetero-protein complexes in the cytosol and also in the membrane of stored RBC. Although the presence of NAC in the storage solution was unable to avoid the formation of thiol-mediated protein complexes, it partially restored the capacity of the cell to metabolize H2O2, indicating the potential use of NAC as an additive in the preservation solution to improve RBC performance after transfusion.

  19. The heritability of metabolite concentrations in stored human red blood cells.

    PubMed

    van 't Erve, Thomas J; Wagner, Brett A; Martin, Sean M; Knudson, C Michael; Blendowski, Robyn; Keaton, Mignon; Holt, Tracy; Hess, John R; Buettner, Garry R; Ryckman, Kelli K; Darbro, Benjamin W; Murray, Jeffrey C; Raife, Thomas J

    2014-08-01

    The degeneration of red blood cells (RBCs) during storage is a major issue in transfusion medicine. Family studies in the 1960s established the heritability of the RBC storage lesion based on poststorage adenosine triphosphate (ATP) concentrations. However, this critical discovery has not been further explored. In a classic twin study we confirmed the heritability of poststorage ATP concentrations and established the heritability of many other RBC metabolites. ATP concentrations and metabolomic profiles were analyzed in RBC samples from 18 twin pairs. On samples stored for 28 days, the heritability of poststorage ATP concentrations were 64 and 53% in CP2D- and AS-3-stored RBCs, respectively. Metabolomic analyses identified 87 metabolites with an estimated heritability of 20% or greater. Thirty-six metabolites were significantly correlated with ATP concentrations (p ≤ 0.05) and 16 correlated with borderline significance (0.05 ≤ p ≤ 0.10). Of the 52 metabolites that correlated significantly with ATP, 24 demonstrated 20% or more heritability. Pathways represented by heritable metabolites included glycolysis, membrane remodeling, redox homeostasis, and synthetic and degradation pathways. We conclude that many RBC metabolite concentrations are genetically influenced during storage. Future studies of key metabolic pathways and genetic modifiers of RBC storage could lead to major advances in RBC storage and transfusion therapy. © 2014 AABB.

  20. Isolation of Leptospira from blood culture bottles.

    PubMed

    Girault, Dominique; Soupé-Gilbert, Marie-Estelle; Geroult, Sophie; Colot, Julien; Goarant, Cyrille

    2017-01-31

    With the increasing use of real-time PCR techniques, Leptospira isolation has mostly been abandoned for the diagnosis of human leptospirosis. However, there is a great value of collecting Leptospira isolates to better understand the epidemiology of this complex zoonosis and to provide the researchers with different isolates. In this study, we have successfully isolated different Leptospira strains from BacT/Alert aerobic blood culture bottles and suggest that this privileged biological material offers an opportunity to isolate leptospires.

  1. Blood culture of the canine patient.

    PubMed

    Hirsh, D C; Jang, S S; Biberstein, E L

    1984-01-15

    Blood for bacteriologic culture was obtained from 581 sick dogs. Of these, 134 (23%) were considered to have bacteremia. The conditions most frequently associated with bacteremia were malignant neoplasms and infections of the skeletal, cardiovascular, and urogenital systems. The most frequently isolated bacteria were members of the family Enterobacteriaceae and coagulase-positive staphylococci, in sum accounting for more than 50% of the 150 isolates. Most of the dogs with bacteremia had high proportions of immature neutrophils, segmented neutrophils, and monocytes in blood. Dogs with bacteremia and osteomyelitis due to staphylococci had normal hemograms. Blood from dogs with bacteremia due to gram-negative bacteria was more likely to have a high proportion of immature and segmented neutrophil leukocytes than was blood from dogs with bacteremia due to a gram-positive species. Toxic neutrophils were observed more often in blood obtained from patients with bacteremia due to gram-negative bacteria. The development of fever correlated with the bacteremic state regardless of the species of bacteria in the blood.

  2. Initial safety and feasibility of cold-stored uncrossmatched whole blood transfusion in civilian trauma patients.

    PubMed

    Yazer, Mark H; Jackson, Byron; Sperry, Jason L; Alarcon, Louis; Triulzi, Darrell J; Murdock, Alan D

    2016-07-01

    The transfusion of cold-stored uncrossmatched whole blood (WB) has not been extensively used in civilian trauma resuscitation. This report details the initial experience with the safety and feasibility of using WB in this setting after a change of practice at a Level 1 trauma center was instituted. Up to two units of uncrossmatched group O positive WB that was leukoreduced using a platelet-sparing filter from male donors were transfused to male trauma patients with hypotension secondary to bleeding. Hemolytic marker haptoglobin and reports of transfusion reactions in these patients were followed. Additionally, transfusion volumes and outcomes were compared to a historical cohort of male trauma patients who received at least one red blood cell (RBC) unit, but not WB, during the first 24 hours of admission. There were 47 WB patients who were transfused with a mean (SD) of 1.74 (0.61) WB units. The median haptoglobin concentration on post-WB transfusion Day 1 was 25.1 (9.3) mg/dL in 7 of 30 non-group O recipients. No adverse reactions in temporal relation to the WB transfusions were reported. There were 145 male historical control patients identified who were resuscitated with component therapy; the median volume of incompatible plasma transfused to the WB versus component therapy group was not significantly different (1,000 vs. 800 mL, respectively; p = 0.38); the mean plasma:RBC (0.99 [0.47] vs. 0.77 [ 0.73], respectively; p = 0.006) and platelet:RBC (0.72 [0.40] vs. 0.51 [0.734], respectively; p < 0.0001) ratios were significantly higher in the WB group. Transfusion of two units of cold-stored uncrossmatched WB is feasible and seems to be safe in civilian trauma resuscitation. Determining the efficacy of WB with regard to reducing the number of blood products transfused in the first 24 hours or improving recipient survival will require a larger randomized trial. Therapeutic study, level IV.

  3. Bone contamination and blood culture in tissue donors.

    PubMed

    Segur, Josep M; Almela, Manuel; Farinas, Oscar; Lazaro, Alexandre; Navarro, Aurora; Trias, Esteve; Domingo, Anna; Marco, Francesc

    2005-01-01

    Swab cultures are the most usual method to detect graft contamination; nevertheless it has been confirmed his limited sensibility. We have studied the relationship between blood cultures, swab surface cultures and cultures of entirely samples of cancellous bone. We have evaluated 5 donors with positive blood culture, from 70 multiorganic donors during 2002. Blood samples were obtained prior the heart arrest. The bone procurement was done just after the organ recovery under aseptic conditions, and surface cultures were performed of each bone. After storage at -80 degrees C, cancellous samples were obtained by trephine and were completely cultured. In one case, the same microorganism grown in blood culture, in 2 of 9 surface cultures, and in 15 of 26 samples of cancellous bone. We conclude that to guarantee allograft's safety it is recommended to add donor's blood culture to the habitual surface swab culture if secondary sterilisation is not performed.

  4. Utility of collecting blood cultures through newly inserted intravenous catheters.

    PubMed

    Isaacman, D J; Karasic, R B

    1990-11-01

    We prospectively examined the utility of obtaining blood cultures through newly inserted intravenous catheters in 99 children who required both a blood culture and placement of an intravenous catheter. Two blood cultures were collected from each patient, one through a freshly inserted intravenous catheter and another through a butterfly needle at a separate venipuncture site. A standardized technique of skin preparation with povidone-iodine was used. The rate of contamination was 1.0% (95% confidence intervals, 0 to 3.0%) for each method. Ten patients had blood cultures yielding true pathogens; in five of these bacteremic children, only one of two sets of blood cultures was positive. We conclude that blood cultures can be collected through freshly placed intravenous catheters without increasing the risk of contamination. These results also raise the possibility that obtaining two blood cultures instead of a single culture may improve the detection of bacteremia in children.

  5. Transfusion of stored red blood cells in critically ill trauma patients: a retrospective study.

    PubMed

    Spadaro, S; Reverberi, R; Fogagnolo, A; Ragazzi, R; Napoli, N; Marangoni, E; Bellini, T; Volta, C A

    2015-01-01

    The many published studies on the effects of the transfusion of stored red blood cells on clinical outcomes yielded discordant results. Therefore, we chose to study patients with severe trauma. The clinical outcomes considered included in-hospital mortality, the occurrence of sepsis, length of stay in intensive care unit and in hospital, and days of mechanical ventilation. We selected all patients with traumatic injury, who received at least 2 red cell units in the first day of admission. Patients were divided into two groups: those who had received fresh red cells only (fresh group) and those who had received at least one "old" red cell unit (old group). The red cells were considered fresh if they had been stored <14 days. The fresh and old groups included 376 and 321 patients, respectively. Baseline demographic and clinical characteristics were comparable between the groups. However, old group received more red cell and plasma units during whole hospital stay (red cells: 11 ± 7 vs 6 ± 4, p < 0.001; plasma: 7 [0-9] vs 3 [0-6]). Among outcomes, only length of stay in intensive care unit (old vs fresh: 18 ± 9 vs 12 ± 8 days, p < 0.001) and in hospital (77 ± 35 vs 45 ± 30 days, p < 0.001) differed significantly between groups. The association remained statistically significant in a multivariate analysis including known confounding factors. Patients with major trauma transfused with old (≥14 days) red cells had a longer length of stay in intensive care unit and in hospital, without any difference in mortality, occurence of sepsis or days of mechanical ventilation.

  6. CO2 -dependent metabolic modulation in red blood cells stored under anaerobic conditions.

    PubMed

    Dumont, Larry J; D'Alessandro, Angelo; Szczepiorkowski, Zbigniew M; Yoshida, Tatsuro

    2016-02-01

    Anaerobic red blood cell (RBC) storage reduces oxidative damage, maintains adenosine triphosphate (ATP) and 2,3-diphosphoglycerate (DPG) levels, and has superior 24-hour recovery at 6 weeks compared to standard storage. This study will determine if removal of CO2 during O2 depletion by gas exchange may affect RBCs during anaerobic storage. This is a matched three-arm study (n = 14): control, O2 and CO2 depleted with Ar (AN), and O2 depleted with 95%Ar/5%CO2 (AN[CO2 ]). RBCs in additives AS-3 or OFAS-3 were evenly divided into three bags, and anaerobic conditions were established by gas exchange. Bags were stored at 1 to 6°C in closed chambers under anaerobic conditions or ambient air, sampled weekly for up to 9 weeks for a panel of in vitro tests. A full metabolomics screening was conducted for the first 4 weeks of storage. Purging with Ar (AN) results in alkalization of the RBC and increased glucose consumption. The addition of 5% CO2 to the purging gas prevented CO2 loss with an equivalent starting and final pH and lactate to control bags (p > 0.5, Days 0-21). ATP levels are higher in AN[CO2 ] (p < 0.0001). DPG was maintained beyond 2 weeks in the AN arm (p < 0.0001). Surprisingly, DPG was lost at the same rate in both control and AN[CO2 ] arms (p = 0.6). Maintenance of ATP in the AN[CO2 ] arm demonstrates that ATP production is not solely a function of the pH effect on glycolysis. CO2 in anaerobic storage prevented the maintenance of DPG, and DPG production appears to be pH dependent. CO2 as well as O2 depletion provides metabolic advantage for stored RBCs. © 2015 AABB.

  7. Seasonal variation in blood and muscle oxygen stores attributed to diving behavior, environmental temperature and pregnancy in a marine predator, the California sea lion.

    PubMed

    Villegas-Amtmann, Stella; Atkinson, Shannon; Paras-Garcia, Alberto; Costa, Daniel P

    2012-08-01

    Survival depends on an animal's ability to find and acquire prey. In diving vertebrates, this ability is directly related to their physiological capability (e.g. oxygen stores). We studied the seasonal variation in oxygen stores, body temperature and body condition in California sea lions (Zalophus californianus) (CSL) as a function of seasonal variation in temperature, primary productivity, diving behavior and reproductive stage. During summer, blood oxygen stores were significantly greater and muscle oxygen stores were significantly lower than in winter. Total oxygen stores, body condition and body temperature did not change between seasons but variations in body temperature were greater during summer. Changes in oxygen stores are partly attributed to diving behavior, temperature and pregnancy that could increase oxygen consumption. Blood and muscle oxygen stores appear to be influenced by reproductive state. Blood oxygen stores are more likely influenced by diving behavior and temperature than muscle oxygen stores. Copyright © 2012 Elsevier Inc. All rights reserved.

  8. Effect of Storage Temperature on Cultured Epidermal Cell Sheets Stored in Xenobiotic-Free Medium

    PubMed Central

    Jackson, Catherine; Aabel, Peder; Eidet, Jon R.; Messelt, Edward B.; Lyberg, Torstein; von Unge, Magnus; Utheim, Tor P.

    2014-01-01

    Cultured epidermal cell sheets (CECS) are used in regenerative medicine in patients with burns, and have potential to treat limbal stem cell deficiency (LSCD), as demonstrated in animal models. Despite widespread use, short-term storage options for CECS are limited. Advantages of storage include: flexibility in scheduling surgery, reserve sheets for repeat operations, more opportunity for quality control, and improved transportation to allow wider distribution. Studies on storage of CECS have thus far focused on cryopreservation, whereas refrigeration is a convenient method commonly used for whole skin graft storage in burns clinics. It has been shown that preservation of viable cells using these methods is variable. This study evaluated the effect of different temperatures spanning 4°C to 37°C, on the cell viability, morphology, proliferation and metabolic status of CECS stored over a two week period in a xenobiotic–free system. Compared to non-stored control, best cell viability was obtained at 24°C (95.2±9.9%); reduced cell viability, at approximately 60%, was demonstrated at several of the temperatures (12°C, 28°C, 32°C and 37°C). Metabolic activity was significantly higher between 24°C and 37°C, where glucose, lactate, lactate/glucose ratios, and oxygen tension indicated increased activation of the glycolytic pathway under aerobic conditions. Preservation of morphology as shown by phase contrast and scanning electron micrographs was best at 12°C and 16°C. PCNA immunocytochemistry indicated that only 12°C and 20°C allowed maintenance of proliferative function at a similar level to non-stored control. In conclusion, results indicate that 12°C and 24°C merit further investigation as the prospective optimum temperature for short-term storage of cultured epidermal cell sheets. PMID:25170754

  9. Detection of Neisseria meningitidis from negative blood cultures and cerebrospinal fluid with the FilmArray blood culture identification panel.

    PubMed

    Pardo, Joe; Klinker, Kenneth P; Borgert, Samuel J; Butler, Brittany M; Rand, Kenneth H; Iovine, Nicole M

    2014-06-01

    The FilmArray blood culture identification (BCID) panel is a rapid molecular diagnostic test approved for use with positive blood culture material. We describe a fatal case of meningococcemia with central nervous system (CNS) involvement detected using the BCID test with culture-negative blood and cerebrospinal fluid.

  10. Adverse Effects of Hemorrhagic Shock Resuscitation with Stored Blood are Ameliorated by Inhaled Nitric Oxide in Lambs

    PubMed Central

    Baron, David M.; Beloiartsev, Arkadi; Nakagawa, Akito; Martyn, Trejeeve; Stowell, Christopher P.; Malhotra, Rajeev; Mayeur, Claire; Bloch, Kenneth D.; Zapol, Warren M.

    2013-01-01

    Objectives Transfusion of stored red blood cells (RBCs) is associated with increased morbidity and mortality in trauma patients. Plasma hemoglobin scavenges nitric oxide (NO), which can cause vasoconstriction, induce inflammation and activate platelets. We hypothesized that transfusion of RBCs stored for prolonged periods would induce adverse effects (pulmonary vasoconstriction, tissue injury, inflammation, and platelet activation) in lambs subjected to severe hemorrhagic shock, and that concurrent inhalation of NO would prevent these adverse effects. Design Animal study. Setting Research laboratory at the Massachusetts General Hospital, Boston, MA. Subjects Seventeen awake Polypay-breed lambs. Interventions Lambs were subjected to 2 h of hemorrhagic shock by acutely withdrawing 50% of their blood volume. Lambs were resuscitated with autologous RBCs stored for 2 h or less (fresh) or 39±2 (mean±SD) days (stored). Stored RBCs were administered with or without breathing NO (80 ppm) during resuscitation and for 21 h thereafter. Measurements and Main Results We measured hemodynamic and oxygenation parameters, markers of tissue injury and inflammation, plasma hemoglobin concentrations, and platelet activation. Peak pulmonary arterial pressure was higher after resuscitation with stored than with fresh RBCs (24±4 vs. 14±2 mmHg, p<0.001) and correlated with peak plasma hemoglobin concentrations (R2=0.56, p=0.003). At 21 h after resuscitation, pulmonary myeloperoxidase activity was higher in lambs resuscitated with stored than with fresh RBCs (11±2 vs. 4±1 U/g, p=0.007). Furthermore, transfusion of stored RBCs increased plasma markers of tissue injury and sensitized platelets to adenosine diphosphate activation. Breathing NO prevented the pulmonary hypertension, and attenuated the pulmonary myeloperoxidase activity, as well as tissue injury and sensitization of platelets to adenosine diphosphate. Conclusions Our data suggest that resuscitation of lambs from hemorrhagic

  11. Diagnosis of brucellosis by using blood cultures.

    PubMed Central

    Ruiz, J; Lorente, I; Pérez, J; Simarro, E; Martínez-Campos, L

    1997-01-01

    The performances of three blood culture systems, Hemoline performance diphasic medium (bioMérieux, Marcy l'Etoile, France), Bactec Plus Aerobic/F* (Becton Dickinson, Paramus, N.J.), and Vital Aer (bioMérieux), were compared for the diagnosis of 17 cases of brucellosis. By using a 5-day incubation protocol, positive results were 52.9, 82.4, and 11.8%, respectively. When the protocol was extended to 7 days, the results were 76.5, 94.1, and 47.1%, respectively. Bactec was the fastest system (P < 0.05). PMID:9276429

  12. Optical Tweezers as a New Biomedical Tool to Measure Zeta Potential of Stored Red Blood Cells

    PubMed Central

    Silva, Carlos A. L.; Fernandes, Heloise P.; Filho, Milton M.; Lucena, Sheyla C.; Costa, Ana Maria D. N.; Cesar, Carlos L.; Barjas-Castro, Maria L.; Santos, Beate S.; Fontes, Adriana

    2012-01-01

    During storage, red blood cells (RBCs) for transfusion purposes suffer progressive deterioration. Sialylated glycoproteins of the RBC membrane are responsible for a negatively charged surface which creates a repulsive electrical zeta potential. These charges help prevent the interaction between RBCs and other cells, and especially among each RBCs. Reports in the literature have stated that RBCs sialylated glycoproteins can be sensitive to enzymes released by leukocyte degranulation. Thus, the aim of this study was, by using an optical tweezers as a biomedical tool, to measure the zeta potential in standard RBCs units and in leukocyte reduced RBC units (collected in CPD-SAGM) during storage. Optical tweezers is a sensitive tool that uses light for measuring cell biophysical properties which are important for clinical and research purposes. This is the first study to analyze RBCs membrane charges during storage. In addition, we herein also measured the elasticity of RBCs also collected in CPD-SAGM. In conclusion, the zeta potential decreased 42% and cells were 134% less deformable at the end of storage. The zeta potential from leukodepleted units had a similar profile when compared to units stored without leukoreduction, indicating that leukocyte lyses were not responsible for the zeta potential decay. Flow cytometry measurements of reactive oxygen species suggested that this decay is due to membrane oxidative damages. These results show that measurements of zeta potentials provide new insights about RBCs storage lesion for transfusion purposes. PMID:22363729

  13. Platelets in blood stored in untreated and siliconed glass bottles and plastic bags. I. Studies in vitro.

    PubMed

    KISSMEYER-NIELSEN, F

    1961-11-01

    The number and function (clot retraction) of platelets in blood stored at 4 degrees C. for three weeks in plastic bags and in untreated and siliconed glass containers were determined using EDTA and acid-citrate dextrose (ACD) as anticoagulants. No essential differences were found.

  14. Urinary di-(2-ethylhexyl) phthalate metabolites for detecting transfusion of autologous blood stored in plasticizer-free bags.

    PubMed

    Leuenberger, Nicolas; Barras, Laura; Nicoli, Raul; Robinson, Neil; Baume, Norbert; Lion, Niels; Barelli, Stefano; Tissot, Jean-Daniel; Saugy, Martial

    2016-03-01

    Autologous blood transfusion (ABT) efficiently increases sport performance and is the most challenging doping method to detect. Current methods for detecting this practice center on the plasticizer di(2-ethlyhexyl) phthalate (DEHP), which enters the stored blood from blood bags. Quantification of this plasticizer and its metabolites in urine can detect the transfusion of autologous blood stored in these bags. However, DEHP-free blood bags are available on the market, including n-butyryl-tri-(n-hexyl)-citrate (BTHC) blood bags. Athletes may shift to using such bags to avoid the detection of urinary DEHP metabolites. A clinical randomized double-blinded two-phase study was conducted of healthy male volunteers who underwent ABT using DEHP-containing or BTHC blood bags. All subjects received a saline injection for the control phase and a blood donation followed by ABT 36 days later. Kinetic excretion of five urinary DEHP metabolites was quantified with liquid chromatography coupled with tandem mass spectrometry. Surprisingly, considerable levels of urinary DEHP metabolites were observed up to 1 day after blood transfusion with BTHC blood bags. The long-term metabolites mono-(2-ethyl-5-carboxypentyl) phthalate and mono-(2-carboxymethylhexyl) phthalate were the most sensitive biomarkers to detect ABT with BTHC blood bags. Levels of DEHP were high in BTHC bags (6.6%), the tubing in the transfusion kit (25.2%), and the white blood cell filter (22.3%). The BTHC bag contained DEHP, despite being labeled DEHP-free. Urinary DEHP metabolite measurement is a cost-effective way to detect ABT in the antidoping field even when BTHC bags are used for blood storage. © 2015 AABB.

  15. Impact of constant storage temperatures and multiple warming cycles on the quality of stored red blood cells.

    PubMed

    Wagner, T; Pabst, M-A; Leitinger, G; Reiter, U; Kozma, N; Lanzer, G; Huppertz, B

    2014-01-01

    Red blood cells (RBCs) are routinely stored in liquid state at temperatures below 6°C, and RBC unit core temperature should not exceed 10°C during transport. Since the critical temperature of 10°C was chosen mostly arbitrarily, this study investigated the effect of both constant temperature settings as well as multiple rewarming cycles on stored RBCs with respect to morphology, biochemical parameters and haemolysis. Buffy coat-depleted filtered RBCs were used as standard products. RBCs were stored at 1-6°C (reference group, n = 12), 13 and 22°C (test groups, n = 12 each) or stored at 1-6°C and warmed up five times to 10, 13, or 22°C for a period of 24 h each. Various biochemical parameters were measured weekly. RBCs were further investigated using electron microscopy. Red blood cells stored constantly at 13 or 22°C showed stable haemolysis rates until day 28 and day 14, respectively. RBCs stored at 1-6°C with five warming-up periods to 10, 13 or 22°C each lasting 24 h (total 120 h) did not exceed the limit of the haemolysis rate at the end of storage. Differently shaped erythrocytes were found in all samples, but more crenate erythrocytes appeared after 42 days of storage independent of temperature profiles. Red cells can be kept at constant temperatures above 6°C without apparent harmful effects at least until day 14, whereas multiple warming cycles for no longer than 24 h at 10, 13 or 22°C with subsequent cooling do not cause quality loss as assessed using the in vitro assays employed in this study. © 2013 International Society of Blood Transfusion.

  16. Effect of additive solutions on red blood cell (RBC) membrane properties of stored RBCs prepared from whole blood held for 24 hours at room temperature.

    PubMed

    Veale, Margaret F; Healey, Gerry; Sparrow, Rosemary L

    2011-01-01

    The quality of RBC components is influenced by collection, processing and storage conditions. Regulations require that whole blood (WB) units be refrigerated within 8 hours and processed into RBCs within 24 hours of collection. Overnight room temperature hold of WB has logistical advantages, but the effect on RBC quality has not been fully investigated. RBC additive solutions were compared for their ability to provide improved quality of RBCs prepared from WB held at room temperature for 24 hours. Leukocyte-reduced RBCs were prepared from WB held at 20°C on cooling plates for 24 hours prior to processing. RBCs were stored in additive solutions, SAG-M (control), Erythrosol-4, and PAGGSM, under standard blood banking conditions and sampled during 49 days of storage. Stored RBCs were evaluated for RBC shape and microparticle (MP) accumulation using flow cytometry. Osmotic fragility, adhesion of RBCs to endothelium under shear stress conditions (0.5 dyne/cm(2) ), and routine RBC quality parameters were assessed. RBCs stored in Erythrosol-4 and PAGGSM had decreased cell size, reduced osmotic fragility, and decreased accumulation of glycophorin A-positive MPs and annexin V-binding MPs compared with RBCs stored in SAG-M. RBCs stored in erythrosol-4 had increased adherence to endothelium at days 42 and 49 compared with RBCs stored in SAG-M or PAGGSM. RBCs stored in PAGGSM or Erythrosol-4 had improved retention of RBC membrane and osmotic resilience. The development of new additive solutions may offer improved quality of RBC components prepared from WB held overnight at room temperature. © 2010 American Association of Blood Banks.

  17. Bartonella henselae Infective Endocarditis Detected by a Prolonged Blood Culture

    PubMed Central

    Mito, Tsutomu; Hirota, Yusuke; Suzuki, Shingo; Noda, Kazutaka; Uehara, Takanori; Ohira, Yoshiyuki; Ikusaka, Masatomi

    2016-01-01

    A 65-year-old Japanese man was admitted with a 4-month history of fatigue and exertional dyspnea. Transthoracic echocardiography revealed a vegetation on the aortic valve and severe aortic regurgitation. Accordingly, infective endocarditis and heart failure were diagnosed. Although a blood culture was negative on day 7 after admission, a prolonged blood culture with subculture was performed according to the patient's history of contact with cats. Consequently, Bartonella henselae was isolated. Bartonella species are fastidious bacteria that cause blood culture-negative infective endocarditis. This case demonstrates that B. henselae may be detected by prolonged incubation of blood cultures. PMID:27746451

  18. Bartonella henselae Infective Endocarditis Detected by a Prolonged Blood Culture.

    PubMed

    Mito, Tsutomu; Hirota, Yusuke; Suzuki, Shingo; Noda, Kazutaka; Uehara, Takanori; Ohira, Yoshiyuki; Ikusaka, Masatomi

    A 65-year-old Japanese man was admitted with a 4-month history of fatigue and exertional dyspnea. Transthoracic echocardiography revealed a vegetation on the aortic valve and severe aortic regurgitation. Accordingly, infective endocarditis and heart failure were diagnosed. Although a blood culture was negative on day 7 after admission, a prolonged blood culture with subculture was performed according to the patient's history of contact with cats. Consequently, Bartonella henselae was isolated. Bartonella species are fastidious bacteria that cause blood culture-negative infective endocarditis. This case demonstrates that B. henselae may be detected by prolonged incubation of blood cultures.

  19. The Relationship of Oxidation Sensitivity of Red Blood Cells and Carbonic Anhydrase Activity in Stored Human Blood: Effect of Certain Phenolic Compounds

    PubMed Central

    Şekeroğlu, Mehmet Ramazan; Balahoroğlu, Ragıp; Karakoyun, Tahsin; Çokluk, Erdem

    2016-01-01

    It has been reported that many modifications occur with the increase of oxidative stress during storage in erythrocytes. In order to delay these negative changes, we evaluated whether the addition of substances likely to protect antioxidant capacity in stored blood would be useful. Therefore, we investigated the effects of resveratrol, tannic acid, and caffeic acid in lipid peroxidation and antioxidant capacity of erythrocytes in stored blood. Donated blood was taken into four CPD containing blood bags. One bag was used as the control, and the others were supplemented with caffeic acid (30 μg/mL), resveratrol (30 μg/mL), and tannic acid (15 μg/mL), respectively. Erythrocyte lipid peroxidation, sensitivity to oxidation, glutathione levels and carbonic anhydrase, glutathione peroxidase, and catalase activities were measured on days 0, 7, 14, 21, and 28. In the control group, erythrocyte malondialdehyde levels and sensitivity to oxidation were increased whereas glutathione, glutathione peroxidase, and catalase levels were decreased (p < 0.05). Resveratrol and caffeic acid prevented malondialdehyde accumulation and preserved glutathione, glutathione peroxidase, and catalase activities in erythrocytes. We demonstrated that resveratrol, caffeic acid, and tannic acid in stored blood could decrease the sensitivity to oxidation of erythrocytes in vitro but did not exhibit such effects on CA activity. PMID:27413740

  20. [Clinical consideration of coagulase negative Staphylococci isolated in blood culture].

    PubMed

    Oshitani, Yohei; Ishikawa, Tomoyuki; Murata, Ken; Aoyagi, Yoshiki; Yabe, Yasuyo; Aoshima, Masahiro

    2012-01-01

    Despite blood culture's usefulness in antimicrobial therapy, fewer blood cultures and the infrequency of more than 1 set in cultures appear to be problems in Japan. Since June 2007 infection control team (ICT) recommended more than 1 set in blood sampling and intervention in positive blood culture, coagulase negative Staphylococci (CNS) has frequently been isolated from blood culture and its clinical significance is often difficult to judge. To determine the effect of ICT intervention, we evaluated the number of blood culture specimens, the frequency of more than 1 set in all blood culture specimens, and decision-making on antimicrobial treatment for CNS isolated retrospectively from blood. The study was divided into term I in August 2007 to July 2008, term II in August 2008 to July 2009, and term III in August 2009 to February 2010. We also analyzed how physicians treated infection or its suspicion after CNS and its drug susceptibility. The monthly number of blood culture specimens increased from 40.3 to 51.6 between terms I and III. The frequency of more than 1 set in a single blood culture session rose significantly from 67% to 89% between these terms (p < 0.001). The number of indeterminate also dropped cases significantly during these 2 terms from 27% to 6% (p = 0.017). Infection or suspected infection cases--45 of 49--had central vein catheter implantation. Inappropriate treatment by physicians in these cases also dropped significantly from 85% (11/13) to 45% (5/11) (p = 0.043) during the same 2 terms. ICT Intervention may thus increase the number of blood culture specimens, enable more than 1 set in blood sampling, make it easier to judge the presence of infection, and increase appropriate treatment by physicians. We thus believe that the quality of antimicrobial treatment could be improved through education such as ICT action.

  1. Systems biology of stored blood cells: can it help to extend the expiration date?

    PubMed

    Paglia, Giuseppe; Palsson, Bernhard Ø; Sigurjonsson, Olafur E

    2012-12-05

    With increasingly stringent regulations regarding deferral and elimination of blood donors it will become increasingly important to extend the expiration date of blood components beyond the current allowed storage periods. One reason for the storage time limit for blood components is that platelets and red blood cells develop a condition called storage lesions during their storage in plastic blood containers. Systems biology provides comprehensive bio-chemical descriptions of organisms through quantitative measurements and data integration in mathematical models. The biological knowledge for a target organism can be translated in a mathematical format and used to compute physiological properties. The use of systems biology represents a concrete solution in the study of blood cell storage lesions, and it may open up new avenues towards developing better storage methods and better storage media, thereby extending the storage period of blood components. This article is part of a Special Issue entitled: Integrated omics.

  2. Cultured blood versus donated blood: long-run perspectives of the economy of blood.

    PubMed

    Mercier Ythier, Jean

    2015-01-01

    Recent advances of fundamental research on the in vitro generation of red blood cells (RBCs) from hematopoietic stem cells in the laboratory open new possibilities of the utilization of cultured RBCs in transfusion medicine. We study the economic challenge of the setup and development of the mass industrial production of RBCs in mature transfusion organizations. We argue that: (i) RBC manufacturing could be set up and developed in the short-medium run for the treatment of the small proportion of transfused patients who have a rare blood type or are alloimmunized against blood antigens; (ii) manufactured RBCs could substitute for donated RBCs in the long run if the physical productivity of RBC engineering technology approaches that of bone marrow.

  3. Correlation between mass and volume of collected blood with positivity of blood cultures.

    PubMed

    Neves, Lariessa; Marra, Alexandre Rodrigues; Camargo, Thiago Zinsly Sampaio; dos Santos, Maura Cristina; Zulin, Flávia; da Silva, Patrícia Candido; de Moura, Natália Ariede; Victor, Elivane da Silva; Pasternak, Jacyr; dos Santos, Oscar Fernando Pavão; Edmond, Michael B; Martino, Marines Dalla Valle

    2015-08-28

    The collection of blood cultures is an extremely important method in the management of patients with suspected infection. Microbiology laboratories should monitor blood culture collection. Over an 8-month period we developed a prospective, observational study in an adult Intensive Care Unit (ICU). We correlated the mass contained in the blood vials with blood culture positivity and we also verified the relationship between the mass of blood and blood volume collected for the diagnosis of bloodstream infection (BSI), as well as we explored factors predicting positive blood cultures. We evaluated 345 patients with sepsis, severe sepsis or septic shock for whom blood culture bottles were collected for the diagnosis of BSI. Of the 55 patients with BSI, 40.0% had peripheral blood culture collection only. BSIs were classified as nosocomial in 34.5%. In the multivariate model, the blood culture mass (in grams) remained a significant predictor of positivity, with an odds ratio 1.01 (i.e., for each additional 1 mL of blood collected there was a 1% increase in positivity; 95% CI 1.01-1.02, p = 0.001; Nagelkerke R Square [R(2)] = 0.192). For blood volume collected, the adjusted odds ratio was estimated at 1.02 (95% CI: 1.01-1.03, p < 0.001; R(2) = 0.199). For each set of collected blood cultures beyond one set, the adjusted odds ratio was estimated to be 1.27 (95% CI: 1.14-1.41, p < 0.001; R(2) = 0.221). Our study was a quality improvement project that showed that microbiology laboratories can use the weight of blood culture bottles to determine if appropriate volume has been collected to improve the diagnosis of BSI.

  4. Plasma and lipids from stored packed red blood cells cause acute lung injury in an animal model.

    PubMed Central

    Silliman, C C; Voelkel, N F; Allard, J D; Elzi, D J; Tuder, R M; Johnson, J L; Ambruso, D R

    1998-01-01

    Transfusion-related acute lung injury (TRALI) is a serious complication of hemotherapy. During blood storage, lipids are generated and released into the plasma. In this study, the role of these lipids in TRALI was investigated using an isolated, perfused rat lung model. Rats were pretreated with endotoxin (LPS) or saline in vivo and the lungs were isolated, ventilated, and perfused with saline, or (a) 5% (vol/ vol) fresh human plasma, (b) plasma from stored blood from the day of isolation (D.0) or from the day of outdate (D.42), (c) lipid extracts from D.42 plasma, or (d) purified lysophosphatidylcholines. Lungs from saline or LPS-pretreated rats perfused with fresh (D.0) plasma showed no pulmonary damage as compared with saline perfused controls. LPS pretreatment/D.42 plasma perfusion caused acute lung injury (ALI) manifested by dramatic changes in both pulmonary artery pressure and edema. Incubation of LPS pre-tx rats with mibefradil, a Ca2+ channel blocker, or WEB 2170, a platelet-activating factor (PAF) receptor antagonist, inhibited ALI caused by D.42 plasma. Lung histology showed neutrophil sequestration without ALI with LPS pretreatment/saline or D.0 plasma perfusion, but ALI with LPS pretreatment/D.42 plasma perfusion, and inhibition of D.42 plasma induced ALI with WEB 2170 or mibefradil. A significant increase in leukotriene E4 was present in LPS-pretreated/D.42 plasma-perfused lungs that was inhibited by WEB 2170. Lastly, significant pulmonary edema was produced when lipid extracts of D.42 plasma or lysophosphatidylcholines were perfused into LPS-pretreated lungs. Lipids caused ALI without vasoconstriction, except at the highest dose employed. In conclusion, both plasma and lipids from stored blood produced pulmonary damage in a model of acute lung injury. TRALI, like the adult respiratory distress syndrome, may be the result of two insults: one derived from stored blood and the other from the clinical condition of the patient. PMID:9525989

  5. The miRNA Profile of Platelets Stored in a Blood Bank and Its Relation to Cellular Damage from Storage.

    PubMed

    Pontes, Thaís Brilhante; Moreira-Nunes, Caroline de Fátima Aquino; Maués, Jersey Heitor da Silva; Lamarão, Letícia Martins; de Lemos, José Alexandre Rodrigues; Montenegro, Raquel Carvalho; Burbano, Rommel Mário Rodriguez

    2015-01-01

    Millions of blood products are transfused each year, and many lives are directly affected by transfusion. Platelet concentrate (PC) is one of the main products derived from blood. Even under good storage conditions, PC is likely to suffer cell damage. The shape of platelets changes after 5 to 7 days of storage at 22°C. Taking into consideration that some platelet proteins undergo changes in their shape and functionality during PC storage. Sixteen PC bags were collected and each PC bag tube was cut into six equal pieces to perform experiments with platelets from six different days of storage. Thus, on the first day of storage, 1/6 of the tube was used for miRNA extraction, and the remaining 5/6 was stored under the same conditions until extraction of miRNAs on each the following five days. Samples were sequenced on an Illumina Platform to demonstrate the most highly expressed miRNAs. Three miRNAs, mir127, mir191 and mir320a were validated by real-time quantitative PCR (RQ-PCR) in 100 PC bags tubes. Our method suggests, the use of the miRNAs mir127 and mir320a as biomarkers to assess the "validity period" of PC bags stored in blood banks for long periods. Thus, bags can be tested on the 5th day of storage for the relative expression levels of mir127 and mir320a. Thus, we highlight candidate miRNAs as biomarkers of storage damage that can be used as tools to evaluate the quality of stored PC. The use of miRNAs as biomarkers of damage is unprecedented and will contribute to improved quality of blood products for transfusions.

  6. The miRNA Profile of Platelets Stored in a Blood Bank and Its Relation to Cellular Damage from Storage

    PubMed Central

    Maués, Jersey Heitor da Silva; Lamarão, Letícia Martins; de Lemos, José Alexandre Rodrigues; Montenegro, Raquel Carvalho; Burbano, Rommel Mário Rodriguez

    2015-01-01

    Millions of blood products are transfused each year, and many lives are directly affected by transfusion. Platelet concentrate (PC) is one of the main products derived from blood. Even under good storage conditions, PC is likely to suffer cell damage. The shape of platelets changes after 5 to 7 days of storage at 22°C. Taking into consideration that some platelet proteins undergo changes in their shape and functionality during PC storage. Sixteen PC bags were collected and each PC bag tube was cut into six equal pieces to perform experiments with platelets from six different days of storage. Thus, on the first day of storage, 1/6 of the tube was used for miRNA extraction, and the remaining 5/6 was stored under the same conditions until extraction of miRNAs on each the following five days. Samples were sequenced on an Illumina Platform to demonstrate the most highly expressed miRNAs. Three miRNAs, mir127, mir191 and mir320a were validated by real-time quantitative PCR (RQ-PCR) in 100 PC bags tubes. Our method suggests, the use of the miRNAs mir127 and mir320a as biomarkers to assess the "validity period" of PC bags stored in blood banks for long periods. Thus, bags can be tested on the 5th day of storage for the relative expression levels of mir127 and mir320a. Thus, we highlight candidate miRNAs as biomarkers of storage damage that can be used as tools to evaluate the quality of stored PC. The use of miRNAs as biomarkers of damage is unprecedented and will contribute to improved quality of blood products for transfusions. PMID:26121269

  7. Detection of bacterial contamination and DNA quantification in stored blood units in 2 veterinary hospital blood banks.

    PubMed

    Stefanetti, Valentina; Miglio, Arianna; Cappelli, Katia; Capomaccio, Stefano; Sgariglia, Elisa; Marenzoni, Maria L; Antognoni, Maria T; Coletti, Mauro; Mangili, Vittorio; Passamonti, Fabrizio

    2016-09-01

    Blood transfusions in veterinary medicine have become increasingly more common and are now an integral part of lifesaving and advanced treatment in small and large animals. Important risks associated with transfusion of blood products include the transmission of various infectious diseases. Several guidelines suggest what infectious agents to screen for in canine and feline transfusion medicine. However, while the risk of bacterial contamination of blood products during storage and administration has not been documented in veterinary medicine, it has emerged as a cause of morbidity and mortality in human transfusion medicine. Clinical experience shows that the majority of blood component bacterial contaminations are caused by only a few species. Unlike other types of bacteria, psychrotolerant species like Pseudomonas spp. and Serratia spp. can proliferate during the storage of blood units at 4°C from a very low titer at the time of blood collection to a clinically significant level (> 10(5) CFU/mL) causing clinical sepsis resulting from red blood cell concentrate transfusions in human medicine. The purpose of this report was to describe the detection and quantification procedures applied in 4 cases of bacterial contamination of canine and feline blood units, which suggest the need for further investigations to optimize patients' safety in veterinary transfusion medicine.

  8. 76 FR 51041 - Hemoglobin Standards and Maintaining Adequate Iron Stores in Blood Donors; Public Workshop

    Federal Register 2010, 2011, 2012, 2013, 2014

    2011-08-17

    ...). Nonetheless, some donors, especially repeat and premenopausal female donors, can develop iron deficiency, with... reduce donor deferrals due to low hemoglobin and hematocrit levels and reduce iron deficiency that can result from blood donation. Different strategies to minimize iron deficiency in blood donors (e.g...

  9. [Blood culture positivity: is it pathogen or contaminant?].

    PubMed

    Balıkçı, Ahmet; Belas, Zeliha; Eren Topkaya, Aynur

    2013-01-01

    Blood culture is the gold standard for diagnosis of bloodstream infections. Many studies have shown that rapid isolation and identification of the microorganisms in blood culture and initiation of early antimicrobial therapy are critically important to reduce the mortality rate. It was found that the rate of contamination in blood cultures is increasing with automated systems developed to facilitate the growth of microorganism and tracking positivity. It is more difficult to interpret a positive blood culture result especially in the case of having only one sample bottle. In this study the effect of growth time observed in the automated blood culture systems was evaluated in terms of interpretation of blood culture results as being pathogens or contaminants. A total of 1201 blood cultures tested in BACTEC 9120 (Becton Dickinson, USA) system in Maltepe University Hospital Medical Microbiology Laboratory, Istanbul, Turkey during one-year period were included in the study and growth times were recorded for positive bottles. The decision about the growth as being a pathogen or contamination was made by considering the clinical condition of the patient, the number of positive blood cultures and the results of inflammation markers (white blood cell counts, procalsitonin and CRP levels). Of the blood cultures 290 (24%) yielded positive results and 73% (212/290) of them were evaluated as pathogens, while 27% (78/290) were identified as contaminants. The mean detection time for clinically significant isolates was 17.87 hours and for contaminants was 40.56 hours. The difference between the growth time of pathogens and contaminants was found statistically significant (p< 0.0001). With regard to all positive results, it was detected that 66% of the bacteria grew within the first 24 hours. While 29.6% of the pathogens grew within 12 hours, none of the contaminants grew during that time. The evaluation of growth time among staphylococci in terms of methicillin resistance

  10. The purified vepoloxamer prevents haemolysis in 42-day stored, DEHP/PVC-free red blood cell units

    PubMed Central

    Cancelas, Jose A.; Rugg, Neeta; Nestheide, Shawnagay; Hill, Sarah E.; Emanuele, R. Martin; McKenzie, Douglas S.

    2017-01-01

    Background Use of the plasticiser di(2-ethylhexyl) phthalate (DEHP) in polyvinyl chloride (PVC) blood bags poses a potential dilemma. The presence of DEHP in blood bags has been shown to be beneficial to red blood cells during storage by diminishing haemolysis. However, DEHP use in PVC may be carcinogenic or estrogenising. Vepoloxamer is a poloxamer with rheological and cytoprotective rheological properties and a favourable toxicity profile in clinical trials. We hypothesised that vepoloxamer may be sufficient to replace the plasticiser DEHP to prevent elevated haemolysis while conserving the biochemical and redox potential++ in RBCs stored for up to 42 days. Materials and methods Paired analyses of aliquots from pooled RBC suspensions of ABO identical donors were aseptically split into test storage containers (DEHP/PVC or DEHP-free/ethylene vinyl acetate [EVA]) supplemented with or without vepoloxamer (at concentrations of 0.1, 1, 5 or 7.89 mg/mL) and cold stored for up to 42 days. Results Vepoloxamer significantly prevented the increased haemolysis induced by the absence of DEHP in EVA bags in a dose-dependent manner by days 28 and 42 of storage (approx. 50% reduction of the maximum concentration of vepoloxamer; p<0.001). There was an inverse correlation between the concentration of vepoloxamer used and the haemolysis rate (r2=0.27, p<0.001) and a direct correlation between haemolysis and phosphatidylserine (PS) exposure (r2=0.42; p<0.01). Increased osmotic fragility and shear induced deformability of 42-day stored RBC in EVA bags was significantly corrected by the addition of vepoloxamer. Discussion Vepoloxamer, in a concentration-dependent fashion, is able to partly rescue the increased haemolysis and PS exposure induced by the absence of the commonly used plasticiser DEHP. These results provide initial but strong evidence to support vepoloxamer use to replace DEHP in long-term storage of RBC. PMID:28263175

  11. Impact of 13.56-MHz radiofrequency identification systems on the quality of stored red blood cells.

    PubMed

    Kozma, Noemi; Speletz, Harald; Reiter, Ursula; Lanzer, Gerhard; Wagner, Thomas

    2011-11-01

    Radiofrequency identification (RFID) technology is emerging as one of the most pervasive computing technologies due to its broad applicability. Storage of red blood cells (RBCs) is a routine procedure worldwide. Depending on the additive solution, RBCs can be stored at 4 ± 2°C up to 49 days. To support the decision of discarding or further using a blood product, temperature measurement of each unit could be provided by RFID application. The safety evaluation of RFID devices was demonstrated in a regulatory agency required study. It has been concluded in limit tests that high frequency-based RFID technology performed safely for blood products; therefore, a longer exposure of radiofrequency (RF) energy on blood units was performed in this study to detect any biologic effects in RBC samples. Buffy coat-depleted, in line-filtered RBCs were used as standard products in all tests. Various variables like pH, potassium, glucose, lactate, hemoglobin (Hb), hematocrit, free Hb, and hemolysis rate were measured in a test group with RFID tags placed on their surface and continuously radiated with 13.56-MHz RFID reader radiation for 42 days while stored at 4 ± 2°C and compared to a control group by two-sample t test. In both groups glucose and pH levels decreased while lactate, free Hb, and potassium increased within the expected levels. The hemolysis rate showed increase after the 25th day but remained below the maximum acceptable threshold of 0.8%. It is feasible to implement RFID-enabled processes, without detecting any known biologic effects of longer exposure of RF energy on the quality of RBCs. © 2011 American Association of Blood Banks.

  12. The stability of iso-α-acids and reduced iso-α-acids in stored blood specimens.

    PubMed

    Rodda, Luke N; Gerostamoulos, Dimitri; Drummer, Olaf H

    2014-06-01

    The long-term stability of the iso-α-acids, and three structurally similar but chemically altered iso-α-acids (known as 'reduced iso-α-acids' and consisting of the rho-, tetrahydro- and hexahydro-iso-α-acid groups) were investigated in whole blood. Pools of blank blood spiked with the four beer-specific ingredient congener groups at two different concentration levels were stored at 20°C, 4°C and -20°C; and extracted in duplicate in weeks 1, 3, 5 and 8, using a previously published method. A loss of 15% of the initial concentration was considered to indicate possible instability and losses greater than 30% demonstrated significant losses. The individual analytes within the four iso-α-acid groups were also measured to determine which iso-α-acids were subject to greater degradation and were responsible for the overall group instability. All four iso-α-acid groups showed significant losses after 8 weeks of storage under room temperature conditions in particularly the natural iso-α-acid group where major losses were observed (96% and 85% losses for low and high concentrations, respectively). Some degradation in all iso-α-acid groups were seen at 4°C samples predominantly due to the 'n' analogs of the groups showing an increased instability in blood. The -20°C storage conditions resulted in minimal changes in concentrations of all analytes. Higher than frozen storage temperatures can result in substantial changes on the stability of the iso-α-acid type groups in blood. The aim of this study was to highlight the stabilities of the IAA analytes in order to assist in the interpretation of IAA in stored blood specimens. Copyright © 2014 Elsevier Ireland Ltd. All rights reserved.

  13. Effects of volume and periodicity on blood cultures.

    PubMed Central

    Li, J; Plorde, J J; Carlson, L G

    1994-01-01

    Blood specimens collected for culture by using the high-volume resin-based BACTEC system over an 18-month period at the Seattle Veterans Administration Center were examined in this study. Of 7,783 cultures obtained, 624 were classified as true positives. Patients in this group had between 20 and 60 ml of blood drawn per culture and separated into 10-ml aliquots for incubation. Analysis of the results stratified by cultured volume and time interval between specimen collection accorded yield advantage to culture volume at the maximal amounts tested. No advantage was observed with any particular interval of collection. Increasing cultured volume from 20 to 40 ml increased yield by 19%. Increasing cultured volume from 40 to 60 ml increased yield by an additional 10%. The same effect was seen whether cultures were drawn simultaneously or serially within 24 h. These observations support other reports demonstrating increased yield with increased cultured blood volume. However, they demonstrate increases in yield at volumes much higher than previously considered. In conclusion, this study demonstrates that high-volume blood cultures drawn serially or simultaneously return the best yields. PMID:7852579

  14. Stability of user-friendly blood typing kits stored under typical military field conditions.

    PubMed

    Bienek, Diane R; Chang, Cheow K; Charlton, David G

    2009-10-01

    To help preserve in-theater strength within deployed military units, commercially available, rapid, user-friendly ABO-Rh blood typing kits were evaluated to determine their stability in storage conditions commonly encountered by the warfighter. Methods for environmental exposure testing were based on MIL-STD-810F. When Eldon Home Kits 2511 were exposed to various temperature/relative humidity conditions, the results were comparable to those obtained with the control group and those obtained with industry-standard methods. For the ABO-Rh Combination Blood Typing Experiment Kits, 2 of the exposure treatments rendered them unusable. In addition, a third set of exposure treatments adversely affected the kits, resulting in approximately 30% blood type misclassifications. Collectively, this evaluation of commercial blood typing kits revealed that diagnostic performance can vary between products, lots, and environmental storage conditions.

  15. The quality of fresh-frozen plasma produced from whole blood stored at 4 degrees C overnight.

    PubMed

    Cardigan, Rebecca; Lawrie, Andrew S; Mackie, Ian J; Williamson, Lorna M

    2005-08-01

    The aim of this study was to assess whether the quality of FFP produced from whole blood stored at 4 degrees C overnight is adequate for its intended purpose. Fresh-frozen plasma (FFP) separated from whole blood (n = 60) leukodepleted (LD) after storage at 4 degrees C overnight (18-24 hr from donation, Day 1 FFP) was compared with that LD within 8 hours of donation (Day 0 FFP, the current standard method). In more than 95 percent of Day 1 FFP units, levels of factor (F) II, FV, FVII, FVIII, F IX, FX, FXI, and FXII were greater than 0.50 U per mL except for von Willebrand factor (VWF) antigen and FVIII, where 92 and 87 percent of units, respectively, contained greater than 0.50 IU per mL. Compared with historical data on FFP stored for 8 hours, fibrinogen, FV, FVIII, and FXI were reduced by 12, 15, 23, and 7 percent, respectively, but other factors were not significantly reduced. Levels of VWF-cleaving protease activity were not different between FFP prepared from paired units of blood (n = 3) held for 8 or 24 hours, but were below the reference range in an additional 2 of 6 units held for 24 hours. The activities of protein S, protein C, antithrombin III, and alpha(2)-antiplasmin were reduced by less than 10 percent in Day 1 FFP (n = 20), but with final levels above the lower limit of the normal range in greater than 95 percent of units. Activated FXII antigen was not significantly raised in plasma stored for 18 to 24 hours, but levels of prothrombin fragment 1 + 2 were slightly increased (0.88 ng/mL, 18-24 hr; 0.65 ng/mL, < 8 hr). These data suggest that there is good retention of relevant coagulation factor activity in plasma produced from whole blood stored at 4 degrees C for 18 to 24 hours and that this would be an acceptable product for most patients requiring FFP.

  16. [Time bacterial growth in blood cultures in neonates].

    PubMed

    Mendoza, Luis; Osorio, Miguel; Fernández, Marisol; Henao, Claudia; Arias, Martha; Mendoza, Laura; Manzano, Stefania; Varela, Ana

    2015-01-01

    Sepsis is a major cause of neonatal morbidity and mortality. To detect the time when the bacterial growth curve is evidenced in the blood sample inoculated blood cultures and comparing the times of bacterial growth between Gram negative and Gram positive bacteria, among the types of neonatal sepsis and identifying microorganisms more often isolated from preterm and term. A descriptive study. 114 positive blood cultures from 1,932 blood cultures taken from 01-May-2010 and 31-May-2014 were evaluated. Data were analyzed with Stata(®) 11.0. 5.9% of blood cultures had bacterial growth. The median and interquartile range of Gram negative times of bacterial growth was 11h (10-13h), for Gram positive coagulase-negative Staphylococcus different (CoNS) 12h (12-18h) and CoNS 42h (36-44h). 95.8% of Gram positive and 96% of Gram negative, were the times of bacterial growth≤24h incubation, whereas the 100% CoNS was positive≤62h of incubation. 100% of sepsis by Gram negative and Gram positive no CoNS and 90% of those caused by CoNS are identified in blood cultures in 48h, so we can conclude that to rule out sepsis, an incubation period of 48h in blood cultures is sufficient. Copyright © 2015 Sociedad Chilena de Pediatría. Publicado por Elsevier España, S.L.U. All rights reserved.

  17. Reducing blood culture contamination by a simple informational intervention.

    PubMed

    Roth, A; Wiklund, A E; Pålsson, A S; Melander, E Z; Wullt, M; Cronqvist, J; Walder, M; Sturegård, E

    2010-12-01

    Compared to truly negative cultures, false-positive blood cultures not only increase laboratory work but also prolong lengths of patient stay and use of broad-spectrum antibiotics, both of which are likely to increase antibiotic resistance and patient morbidity. The increased patient suffering and surplus costs caused by blood culture contamination motivate substantial measures to decrease the rate of contamination, including the use of dedicated phlebotomy teams. The present study evaluated the effect of a simple informational intervention aimed at reducing blood culture contamination at Skåne University Hospital (SUS), Malmö, Sweden, during 3.5 months, focusing on departments collecting many blood cultures. The main examined outcomes of the study were pre- and postintervention contamination rates, analyzed with a multivariate logistic regression model adjusting for relevant determinants of contamination. A total of 51,264 blood culture sets were drawn from 14,826 patients during the study period (January 2006 to December 2009). The blood culture contamination rate preintervention was 2.59% and decreased to 2.23% postintervention (odds ratio, 0.86; 95% confidence interval, 0.76 to 0.98). A similar decrease in relevant bacterial isolates was not found postintervention. Contamination rates at three auxiliary hospitals did not decrease during the same period. The effect of the intervention on phlebotomists' knowledge of blood culture routines was also evaluated, with a clear increase in level of knowledge among interviewed phlebotomists postintervention. The present study shows that a relatively simple informational intervention can have significant effects on the level of contaminated blood cultures, even in a setting with low rates of contamination where nurses and auxiliary nurses conduct phlebotomies.

  18. Improving the sensitivity of blood culture for Streptococcus pneumoniae.

    PubMed

    Saha, Samir; Darmstadt, Gary; Naheed, Aliya; Arifeen, Shams; Islam, Maksuda; Fatima, Kaniz; Breiman, Robert; Sack, David; Hamer, Davidson

    2011-06-01

    Isolation of Streptococcus pneumoniae is jeopardized by low sensitivity of blood culture, autolysis and contamination with fast-growing organism(s). We performed an immunochromatographic (ICT) test for S. pneumoniae on chocolatized blood culture bottles and also sub-cultured contaminated bottles on a selective medium, thus identifying an additional eight and three cases, respectively, and improving the detection of pneumococcus by 23% (48% vs. 59%). Prescreening of culture bottles in a blinded fashion could rationalize the use of ICT with ~99% accuracy. These two approaches can aid microbiology laboratories in resource-poor countries to substantially improve rates of detection of S. pneumoniae.

  19. Blood cultures in emergency medical admissions: a key patient cohort.

    PubMed

    Chotirmall, Sanjay H; Callaly, Elizabeth; Lyons, Judith; O'Connell, Brian; Kelleher, Mary; Byrne, Declan; O'Riordan, Deirdre; Silke, Bernard

    2016-02-01

    Blood cultures are performed in the emergency room when sepsis is suspected, and a cohort of patients is thereby identified. The present study investigated the outcomes (mortality and length of hospital stay) in this group following an emergency medical admission. Prospective assessment of all emergency medical admissions presenting to the emergency department at St James's Hospital, Dublin, over an 11-year period (2002-2012) was carried out. Outcomes including 30-day in-hospital mortality and length of stay were explored in the context of an admission blood culture. Generalized estimating equations, logistic or zero-truncated Poisson multivariate models were used, with adjustment for confounding variables including illness severity, comorbidity, and chronic disabling disease, to assess the effect of an urgent blood culture on mortality and length of stay. A total of 60 864 episodes were recorded in 35 168 patients admitted over the time period assessed. Patients more likely to undergo blood cultures in the emergency department were male, younger, and had more comorbidity. Univariate and multivariate analyses showed that those who had a blood culture, irrespective of result, had increased mortality and a longer in-hospital stay. This was highest for those with a positive culture, irrespective of the organism isolated. A clinical decision to request a blood culture identified a subset of emergency admissions with markedly worse outcomes. This patient cohort warrants close monitoring in the emergency setting.

  20. Reducing blood-culture contamination through an education program.

    PubMed

    Robert, Ruth R

    2011-01-01

    A blood culture is the cornerstone of an established etiological diagnosis of septicemia. Although it is not currently possible to eliminate blood-culture contamination, many interventions have been shown to reduce contamination rates. Retrospective data analysis through an initial audit with major departments at one hospital, including the intensive care unit and emergency department, showed that the blood-culture contamination rate was 4.8%, which is more than the set standard (ie, less than 3%). A decrease in blood-culture contamination rates from the initial 4.8% to less than 3% was obtained with a supervised training and evaluation program through collaborative efforts of the nursing and laboratory departments.

  1. Mortality increases after massive exchange transfusion with older stored blood in canines with experimental pneumonia.

    PubMed

    Solomon, Steven B; Wang, Dong; Sun, Junfeng; Kanias, Tamir; Feng, Jing; Helms, Christine C; Solomon, Michael A; Alimchandani, Meghna; Quezado, Martha; Gladwin, Mark T; Kim-Shapiro, Daniel B; Klein, Harvey G; Natanson, Charles

    2013-02-28

    Two-year-old purpose-bred beagles (n = 24) infected with Staphylococcus aureus pneumonia were randomized in a blinded fashion for exchange transfusion with either 7- or 42-day-old canine universal donor blood (80 mL/kg in 4 divided doses). Older blood increased mortality (P = .0005), the arterial alveolar oxygen gradient (24-48 hours after infection; P ≤ .01), systemic and pulmonary pressures during transfusion (4-16 hours) and pulmonary pressures for ~ 10 hours afterward (all P ≤ .02). Further, older blood caused more severe lung damage, evidenced by increased necrosis, hemorrhage, and thrombosis (P = .03) noted at the infection site postmortem. Plasma cell–free hemoglobin and nitric oxide (NO) consumption capability were elevated and haptoglobin levels were decreased with older blood during and for 32 hours after transfusion (all P ≤ .03). The low haptoglobin (r = 0.61; P = .003) and high NO consumption levels at 24 hours (r = −0.76; P < .0001) were associated with poor survival. Plasma nontransferrin-bound and labile iron were significantly elevated only during transfusion (both P = .03) and not associated with survival (P = NS). These data from canines indicate that older blood after transfusion has a propensity to hemolyze in vivo, releases vasoconstrictive cell-free hemoglobin over days, worsens pulmonary hypertension, gas exchange, and ischemic vascular damage in the infected lung, and thereby increases the risk of death from transfusion.

  2. Does endothelin mobilize calcium from intracellular store sites in rat aortic vascular smooth muscle cells in primary culture

    SciTech Connect

    Miasiro, N.; Yamamoto, H.; Kanaide, H.; Nakamura, M.

    1988-10-14

    In the presence of endothelin, there was a rapid increase in the /sup 45/Ca++ efflux from primary cultured rat vascular smooth muscle cells, both in physiological salt solution and in calcium free medium containing 2 mM EGTA. The /sup 45/Ca++ influx was not affected. The endothelin-induced, transient increase in cytosolic calcium concentration is probably mainly due to release of calcium from the intracellular store in vascular smooth muscle cells.

  3. Bacteriological culture of blood from critically ill neonatal calves.

    PubMed Central

    Fecteau, G; Van Metre, D C; Paré, J; Smith, B P; Higgins, R; Holmberg, C A; Jang, S; Guterbock, W

    1997-01-01

    The objectives of this study were to estimate the prevalence of bacteremia in critically ill, neonatal calves with severe diarrhea or depression, and to describe the variety of bacteria involved. Two studies were conducted in the summers of 1991 and 1993 involving 190 neonatal calves, 1-day to 19-days-old. Bacteremia was detected by blood culture in 31% (28/90) of calves in study 1, and in 24% (19/79) of ill calves and 0% (0/21) of control calves in study 2. Bacteria cultured from blood included Escherichia coli (51% of all isolates), other gram-negative enterics (25.5%), gram-negative anaerobes (5.9%), gram-positive cocci (11.8%), and gram-positive rods (5.9%). Among clinically ill calves, the average age was significantly lower in the blood culture-negative group (5.5 d) than in the blood culture-positive group (7.5 d) (P = 0.004). Mean serum IgG concentration was significantly (P = 0.0001) lower in blood culture-positive calves (1.146 g/L) than in blood culture-negative calves (3.077 g/L). The mortality rate was significantly (P < 0.0001) higher in the blood culture-positive group (57.4%) than in the blood culture-negative group (15.1%). Bacteremia appeared to be a frequent entity in this particular rearing situation. Early recognition of the problem, as well as appropriate treatment, may be beneficial in increasing survival rates. Results also support the need to address the failure of passive transfer of maternal antibodies to prevent bacteremia in calves. Images Figure 1. PMID:9028592

  4. Blood culture results do not affect treatment in complicated cellulitis.

    PubMed

    Paolo, William F; Poreda, Andrew R; Grant, William; Scordino, David; Wojcik, Susan

    2013-08-01

    Cellulitis, a frequently encountered complaint in the Emergency Department, is typically managed with antibiotics. There is some debate as to whether obtaining blood cultures and knowing their results would change the management of cellulitis, although most authors argue that information from blood cultures does not change the empirical management of uncomplicated cellulitis. However, for complicated cellulitis (as defined by the presence of significant comorbidity), there is considerable disagreement and lack of evidence as to the utility of blood cultures. Our aim was to determine the role of blood cultures in the management of complicated cellulitis. This retrospective chart review assessed the utility of obtaining blood cultures in complicated cellulitis (as defined by active chemotherapy, dialysis, human immunodeficiency virus/acquired immune deficiency syndrome, diabetes, or organ transplantation) vs. a cohort of individuals without medical comorbidity. Six hundred and thirty-nine patients were identified, 314 of which were deemed cases and 325 controls. Within the cases, 29 of 314 returned as positive blood cultures vs. 17 of 325 positive blood culture controls within the cases (p = 0.05; odds ratio = 1.84; 95% confidence interval 0.99-3.43). A clinically significant change in management (a change in the class of antibiotic) was found in 6 of 314 cases vs. 4 of 325 controls (p = 0.578; odds ratio = 1.5525; 95% confidence interval 0.434-5.5541). Within this cohort of patients with complicated cellulitis, blood cultures rarely changed management from empirical coverage. Copyright © 2013 Elsevier Inc. All rights reserved.

  5. Automated quantitative analysis of 3D morphology and mean corpuscular hemoglobin in human red blood cells stored in different periods.

    PubMed

    Moon, Inkyu; Yi, Faliu; Lee, Yeon H; Javidi, Bahram; Boss, Daniel; Marquet, Pierre

    2013-12-16

    Quantitative phase (QP) images of red blood cells (RBCs), which are obtained by off-axis digital holographic microscopy, can provide quantitative information about three-dimensional (3D) morphology of human RBCs and the characteristic properties such as mean corpuscular hemoglobin (MCH) and MCH surface density (MCHSD). In this paper, we investigate modifications of the 3D morphology and MCH in RBCs induced by the period of storage time for the purpose of classification of RBCs with different periods of storage by using off-axis digital holographic microscopy. The classification of RBCs based on the duration of storage is highly relevant because a long storage of blood before transfusion may alter the functionality of RBCs and, therefore, cause complications in patients. To analyze any changes in the 3D morphology and MCH of RBCs due to storage, we use data sets from RBC samples stored for 8, 13, 16, 23, 27, 30, 34, 37, 40, 47, and 57 days, respectively. The data sets consist of more than 3,300 blood cells in eleven classes, with more than 300 blood cells per class. The classes indicate the storage period of RBCs and are listed in chronological order. Using the RBCs donated by healthy persons, the off-axis digital holographic microscopy reconstructs several quantitative phase images of RBC samples stored for eleven different periods. We employ marker-controlled watershed transform to remove the background in the RBC quantitative phase images obtained by the off-axis digital holographic microscopy. More than 300 single RBCs are extracted from the segmented quantitative phase images for each class. Such a large number of RBC samples enable us to obtain statistical distributions of the characteristic properties of RBCs after a specific period of storage. Experimental results show that the 3D morphology of the RBCs, in contrast to MCH, is essentially related to the aging of the RBCs.

  6. Stored blood--an effective immunosuppressive method for transplantation of kidneys from unrelated donors. An 11-year follow-up.

    PubMed

    Galvão, M M; Peixinho, Z F; Mendes, N F; Sabbaga, E

    1997-06-01

    Thirty-seven patients were submitted to kidney transplantation after transfusion at 2-week intervals with 4-week stored blood from their potential donors. All patients and donors were typed for HLA-A-B and DR antigens. The patients were also tested for cytotoxic antibodies against donor antigens before each transfusion. The percentage of panel reactive antibodies (PRA) was determined against a selected panel of 30 cell donors before and after the transfusions. The patients were immunosuppressed with azathioprine and prednisone. Rejection crises were treated with methylprednisolone. The control group consisted of 23 patients who received grafts from an unrelated donor but who did not receive donor-specific pretransplant blood transfusion. The incidence and reversibility of rejection episodes, allograft loss caused by rejection, and patient and graft survival rates were determined for both groups. Non-parametric methods (chi-square and Fisher tests) were used for statistical analysis, with the level of significance set at P < 0.05. The incidence and reversibility of rejection crises during the first 60 post-transplant days did not differ significantly between groups. The actuarial graft and patient survival rates at five years were 56% and 77%, respectively, for the treated group and 39.8% and 57.5% for the control group. Graft loss due to rejection was significantly higher in the untreated group (P = 0.0026) which also required more intense immunosuppression (P = 0.0001). We conclude that transfusions using stored blood have the immunosuppressive effect of fresh blood transfusions without the risk of provoking a widespread formation of antibodies. In addition, this method permits a reduction of the immunosuppressive drugs during the process without impairing the adequate functioning of the renal graft.

  7. Multiphoton excited hemoglobin fluorescence and third harmonic generation for non-invasive microscopy of stored blood.

    PubMed

    Saytashev, Ilyas; Glenn, Rachel; Murashova, Gabrielle A; Osseiran, Sam; Spence, Dana; Evans, Conor L; Dantus, Marcos

    2016-09-01

    Red blood cells (RBC) in two-photon excited fluorescence (TPEF) microscopy usually appear as dark disks because of their low fluorescent signal. Here we use 15fs 800nm pulses for TPEF, 45fs 1060nm pulses for three-photon excited fluorescence, and third harmonic generation (THG) imaging. We find sufficient fluorescent signal that we attribute to hemoglobin fluorescence after comparing time and wavelength resolved spectra of other expected RBC endogenous fluorophores: NADH, FAD, biliverdin, and bilirubin. We find that both TPEF and THG microscopy can be used to examine erythrocyte morphology non-invasively without breaching a blood storage bag.

  8. Multiphoton excited hemoglobin fluorescence and third harmonic generation for non-invasive microscopy of stored blood

    PubMed Central

    Saytashev, Ilyas; Glenn, Rachel; Murashova, Gabrielle A.; Osseiran, Sam; Spence, Dana; Evans, Conor L.; Dantus, Marcos

    2016-01-01

    Red blood cells (RBC) in two-photon excited fluorescence (TPEF) microscopy usually appear as dark disks because of their low fluorescent signal. Here we use 15fs 800nm pulses for TPEF, 45fs 1060nm pulses for three-photon excited fluorescence, and third harmonic generation (THG) imaging. We find sufficient fluorescent signal that we attribute to hemoglobin fluorescence after comparing time and wavelength resolved spectra of other expected RBC endogenous fluorophores: NADH, FAD, biliverdin, and bilirubin. We find that both TPEF and THG microscopy can be used to examine erythrocyte morphology non-invasively without breaching a blood storage bag. PMID:27699111

  9. The mechanical properties of stored red blood cells measured by a convenient microfluidic approach combining with mathematic model.

    PubMed

    Wang, Ying; You, Guoxing; Chen, Peipei; Li, Jianjun; Chen, Gan; Wang, Bo; Li, Penglong; Han, Dong; Zhou, Hong; Zhao, Lian

    2016-03-01

    The mechanical properties of red blood cells (RBCs) are critical to the rheological and hemodynamic behavior of blood. Although measurements of the mechanical properties of RBCs have been studied for many years, the existing methods, such as ektacytometry, micropipette aspiration, and microfluidic approaches, still have limitations. Mechanical changes to RBCs during storage play an important role in transfusions, and so need to be evaluated pre-transfusion, which demands a convenient and rapid detection method. We present a microfluidic approach that focuses on the mechanical properties of single cell under physiological shear flow and does not require any high-end equipment, like a high-speed camera. Using this method, the images of stretched RBCs under physical shear can be obtained. The subsequent analysis, combined with mathematic models, gives the deformability distribution, the morphology distribution, the normalized curvature, and the Young's modulus (E) of the stored RBCs. The deformability index and the morphology distribution show that the deformability of RBCs decreases significantly with storage time. The normalized curvature, which is defined as the curvature of the cell tail during stretching in flow, suggests that the surface charge of the stored RBCs decreases significantly. According to the mathematic model, which derives from the relation between shear stress and the adherent cells' extension ratio, the Young's moduli of the stored RBCs are also calculated and show significant increase with storage. Therefore, the present method is capable of representing the mechanical properties and can distinguish the mechanical changes of the RBCs during storage. The advantages of this method are the small sample needed, high-throughput, and easy-use, which make it promising for the quality monitoring of RBCs.

  10. Analysis of stored and transplanted cord blood units from KoreaCORD: reappraisal of banking guidelines and selection strategy.

    PubMed

    Lee, Young-Ho; Kwon, Young-Hee; Hwang, Kyoujung; Jun, Hyunju; Lee, Mi-Ae; Jang, Hyung-In; Nah, Jung-Hwa; Koo, Hong-Hoe; Hwang, Tai-Ju

    2013-01-01

    We analyzed the characteristics of stored and transplanted cord blood (CB) units from the Korean network for public CB donation (KoreaCORD) to reassess the banking guidelines and optimize CB selection based on cell dose and human leukocyte antigen (HLA) mismatching. We retrospectively reviewed data, with regard to total nucleated cell (TNC) count and HLA match in the KoreaCORD registry from August 2001 to December 2010. A total of 21,914 CB units have been registered, of which 904 units (4.1%) contained less than 5 × 10(8) TNCs, which did not meet the present storage criteria for public CB banking in Korea. Although the proportion of stored CBs providing TNC of 5 × 10(8) to 7.9 × 10(8) was 45.7%, only 22.0% of all transplanted CBs were derived from these stored CBs. In the single CB transplantation setting, 79% (85/108) of CB units provided 4 × 10(7) TNCs/kg or more in the transplanted one-mismatch (1-MM) CB units and 51% (19/37) of CBs provided 6 × 10(7) TNCs/kg or more in the transplanted 2-MM CB units. The minimal requirement of TNCs for banking of CB units for public banking should be evaluated and increased to support the selection of CB units with higher cell doses, especially for use in the 1- and 2-MM transplant settings. © 2012 American Association of Blood Banks.

  11. Impact of allogeneic 2-RBC apheresis on iron stores of Brazilian blood donors.

    PubMed

    Mendrone, Alfredo; Arrais, Cyntia Araújo; Almeida Neto, César; Gualandro, Sandra de Fátima Menocci; Dorlhiac-Llacer, Pedro Enrique; Chamone, Dalton de Alencar Fischer; Sabino, Ester Cerdeira

    2009-08-01

    One limiting factor for automated two-red blood cells collections (2-RBC) is its potential iron depletion. We analyzed hematological parameters and iron balance before, two and four months after 2-RBC of 96 non-supplemented male donors. Four months after 2-RBC, ferritin level was significantly lower (P<0.01) than baseline levels and the number of donors who presented ferritin <30 ng/ml increased from 18 to 47. We concluded that four months was not sufficient for iron recuperation in the population studied. In an attempt to avoid iron depletion after 2-RBC, we recommend augmentation in the interval between blood donations and pre-donation ferritin measurement.

  12. [Blood culture analysis in children with central venous catheter].

    PubMed

    Delińska-Galińska, Anna; Arłukowicz, Elzbieta; Plata-Nazar, Katarzyna; Luczak, Grazyna; Kozielska, Ewa; Kotłowska-Kmieć, Aldona; Borkowska, Anna

    2008-01-01

    We analyzed 99 blood cultures taken from 28 children with central venous catheter. Children were hospitalized in pediatric, pediatric surgery and pediatric intensive care department. All samples were collected from peripherial vein. Positive blood cultures were obtained more frequently from children with central venous catheter than from children without central venous catheter (57.5% vs. 7.4%). Staphylococcus epidermidis was the most frequently obtained bacteria. The other bacterial species were obtained less frequently. The highest percentage of multi resistant straines was isolated from blood samples collected from intensive care department patients. In each departments in which coagulase-negative Staphylococci were isolated, metycillin-resistant straines dominated.

  13. Rapid Identification of Pathogens from Pediatric Blood Cultures by Use of the FilmArray Blood Culture Identification Panel

    PubMed Central

    Polanco, Wanda; Carter, Donna; Shulman, Stanford

    2014-01-01

    The performance of the FilmArray blood culture identification (BCID) panel has been studied in adult patients. We describe here an evaluation of this assay for the rapid identification of pathogens in Bactec Peds Plus/F and Bactec standard anaerobic/F bottles that contained blood samples from pediatric patients at a tertiary care children's hospital. PMID:25274998

  14. Post mortem concentrations of endogenous gamma hydroxybutyric acid (GHB) and in vitro formation in stored blood and urine samples.

    PubMed

    Busardò, Francesco Paolo; Bertol, Elisabetta; Vaiano, Fabio; Baglio, Giovanni; Montana, Angelo; Barbera, Nunziata; Zaami, Simona; Romano, Guido

    2014-10-01

    Gamma-hydroxybutyrate (GHB) is a central nervous system depressant, primarily used as a recreational drug of abuse with numerous names. It has also been involved in various instances of drug-facilitated sexual assault due to its potential incapacitating effects. The first aim of this paper is to measure the post-mortem concentration of endogenous GHB in whole blood and urine samples of 30 GHB free-users, who have been divided according to the post-mortem interval (PMI) in three groups (first group: 24-36h; second group: 37-72h; third group: 73-192h), trying to evaluate the role of PMI in affecting post mortem levels. Second, the Authors have evaluated the new formation of GHB in vitro in blood and urine samples of the three groups, which have been stored at -20°C, 4°C and 20°C over a period of one month. The concentrations were measured by GC-MS after liquid-liquid extraction according to the method validated and published by Elliot (For. Sci. Int., 2003). For urine samples, GHB concentrations were creatinine-normalized. In the first group the GHB mean concentration measured after autopsy was: 2.14mg/L (range 0.54-3.21mg/L) in blood and 3.90mg/g (range 0.60-4.81mg/g) in urine; in the second group it was: 5.13mg/L (range 1.11-9.60mg/L) in blood and 3.93mg/g (range 0.91-7.25mg/g) in urine; in the third group it was: 11.8mg/L (range 3.95-24.12mg/L) in blood and 9.83mg/g (range 3.67-21.90mg/g) in urine. The results obtained in blood and urine samples showed a statistically significant difference among groups (p<0.001) in the first analysis performed immediately after autopsy. Throughout the period of investigation up to 4 weeks, the comparison of storage temperatures within each group showed in blood and urine samples a mean difference at 20°C compared to -20°C not statistically significant at the 10% level. These findings allow us to affirm that the PMI strongly affects the post mortem production of GHB in blood and urine samples. Regarding the new formation of

  15. Limited Resuscitation With Fresh or Stored Whole Blood Corrects Cardiovascular and Metabolic Function in a Rat Model of Polytrauma and Hemorrhage.

    PubMed

    Chen, Jacob; Wu, Xiaowu; Keesee, Jeffrey; Liu, Bin; Darlington, Daniel N; Cap, Andrew P

    2017-02-01

    We have recently shown that human whole blood stored at 4°C maintains hemostatic and platelet function. In this study, we compared restoration of hemodynamic, metabolic and hemostatic function after limited resuscitation with rat fresh whole blood, rat stored whole blood, or Lactated Ringers in traumatized rats. Rat whole blood was stored for 10 days at 4°C for evaluation of hemostatic function. Polytrauma was performed on isoflurane-anesthetized Sprague-Dawley rats (350-450 g) by damage to the intestines, liver, right leg skeletal muscle, and right femur fracture, followed by 40% hemorrhage. At 1 h, rats were resuscitated (20%) with either fresh whole blood (FWB), stored whole blood, 4°C for 7 days (SWB), Lactated Ringers (LR), or nothing. Blood samples were taken before and 2 h after trauma and hemorrhage to evaluate metabolic and hemostatic function. Whole blood stored for 10 days showed a significant prolongation in prothrombin time (PT) and activated partial thromboplastin time (aPTT), and fall in fibrinogen concentration, but no change in Maximum Clot Firmness or speed of clot formation. Platelet function was maintained until day 7 in storage, than fell significantly. Polytrauma and hemorrhage in rats led to a fall in arterial pressure, plasma bicarbonate, fibrinogen, and platelet function, and a rise in plasma lactate, PT, aPTT, and creatinine. Resuscitation with either FWB or 7 day SWB, but not LR, returned arterial pressure, plasma lactate and plasma bicarbonate to levels similar to control, but had no effect on the fall in fibrinogen or platelet function, or the rise in PT, aPTT, or creatinine. Hemostatic and platelet function of rat whole blood stored at 4°C is preserved for at least 7 days in vitro. Low volume resuscitation with SWB or FWB, but not LR, restores hemodynamic and metabolic function, but not the coagulopathy after severe trauma and hemorrhage.

  16. Quality assurance in blood culture: A retrospective study of blood culture contamination rate in a tertiary hospital in Nigeria

    PubMed Central

    Chukwuemeka, Iregbu Kenneth; Samuel, Yakubu

    2014-01-01

    Background: Blood culture is a critical tool for diagnosing septicaemia. Quite frequently, contamination of blood sample poses a great challenge to accurate diagnosis. This study evaluated the rate of blood culture contamination in our hospital over a one-year period. Materials and Methods: It was a retrospective study of 1032 blood cultures carried out in a clinical laboratory of a tertiary hospital in North Central part of Nigeria between 2010 and 2011. Results: There were 730 blood cultures from paediatric and 302 adult patients. The overall yield was 22%; 107 out of the 730 were contaminated giving a contamination rate of 10.4%. Contamination rate was higher in children than in adult (11% vs 8%) specimen. These rates were much higher than the acceptable benchmark of 2-3%. The main contaminants were coagulase negative Staphylococcus, Bacillus species, Diphtheroids and Enterococcus species. Conclusion: Contamination rate is high, and mainly due to normal skin flora, suggesting aseptic collection challenges as the main cause. We recommend a review of the entire process of blood collection for culture and analysis with a view to instituting appropriate quality assurance measures to reduce the contamination rate. PMID:25013249

  17. Rapid Detection of Polymyxin-Resistant Enterobacteriaceae from Blood Cultures.

    PubMed

    Jayol, Aurélie; Dubois, Véronique; Poirel, Laurent; Nordmann, Patrice

    2016-09-01

    Enterobacterial strains resistant to polymyxins are being increasingly reported worldwide. The conventional methods for detection of colistin-resistant isolates such as broth microdilution remain time-consuming (24 to 48 h), and methods such as disc diffusion and Etest are not reliable. Recently, the rapid polymyxin NP test was developed for rapid identification of polymyxin-resistant Enterobacteriaceae This test is based on the detection of glucose metabolism related to bacterial growth in the presence of a defined concentration of colistin (or polymyxin B). The formation of acid metabolites is evidenced by a color change of a pH indicator (red phenol) in less than 2 h. In this study, the polymyxin NP test was evaluated for detection of colistin-resistant Enterobacteriaceae directly from blood cultures. The test was performed with 73 blood culture sets (either spiked or clinical blood cultures) with various enterobacterial species. The test exhibited excellent discrimination between polymyxin-resistant and polymyxin-susceptible enterobacterial isolates, and results are obtained from blood cultures within 4 h. It is easy to perform and requires neither subculture nor a centrifugation step. This test is rapid, specific, and sensitive and allows early identification of polymyxin-resistant Enterobacteriaceae directly from blood cultures. Copyright © 2016, American Society for Microbiology. All Rights Reserved.

  18. Rapid Detection of Polymyxin-Resistant Enterobacteriaceae from Blood Cultures

    PubMed Central

    Jayol, Aurélie; Dubois, Véronique; Poirel, Laurent

    2016-01-01

    Enterobacterial strains resistant to polymyxins are being increasingly reported worldwide. The conventional methods for detection of colistin-resistant isolates such as broth microdilution remain time-consuming (24 to 48 h), and methods such as disc diffusion and Etest are not reliable. Recently, the rapid polymyxin NP test was developed for rapid identification of polymyxin-resistant Enterobacteriaceae. This test is based on the detection of glucose metabolism related to bacterial growth in the presence of a defined concentration of colistin (or polymyxin B). The formation of acid metabolites is evidenced by a color change of a pH indicator (red phenol) in less than 2 h. In this study, the polymyxin NP test was evaluated for detection of colistin-resistant Enterobacteriaceae directly from blood cultures. The test was performed with 73 blood culture sets (either spiked or clinical blood cultures) with various enterobacterial species. The test exhibited excellent discrimination between polymyxin-resistant and polymyxin-susceptible enterobacterial isolates, and results are obtained from blood cultures within 4 h. It is easy to perform and requires neither subculture nor a centrifugation step. This test is rapid, specific, and sensitive and allows early identification of polymyxin-resistant Enterobacteriaceae directly from blood cultures. PMID:27307457

  19. Biochemical and Cellular Changes in Leukocyte-Depleted Red Blood Cells Stored for Transfusion

    PubMed Central

    Nogueira, Diana; Rocha, Susana; Abreu, Estela; Costa, Elísio; Santos-Silva, Alice

    2015-01-01

    Summary Background To evaluate biochemical and cellular changes associated with the storage of leukocyte-depleted red blood cells (RBCs). Methods We investigated 10 leukocyte-depleted RBC units, randomly chosen from volunteer donors. Every week an aliquot was collected for laboratorial evaluation, which included complete cell blood count, glucose-6-phosphate dehydrogenase (G6PD) activity, extracellular sodium, potassium and pH, membrane-bound hemoglobin (MBH), band 3 profile, and quantification of RBC membrane proteins composition. Results We observed an increase in mean cell volume (from 91.86 ± 4.65 fl to 98.10 ± 5.80 fl, day 0 vs. day 21; p < 0.05), red cell distribution width, percentage of macrocytic RBCs, reticulocyte hemoglobin content and a decreased percentage of microcytic RBCs, mean cell volume concentration and G6PD activity. The extracellular concentration of sodium decreased, and that of potassium increased significantly over time. RBC membrane composition revealed an increase in spectrin/ankyrin ratio after 21 days (from 4.84 ± 0.99 to 5.27 ± 0.94, day 0 vs. day 21; p < 0.05). At day 35, a decrease in ankyrin (from 6.44 ± 1.70% to 5.49 ± 1.96%, day 0 vs. day 35; p < 0.05), in protein 4.1/band 3, protein 4.2/band 3, and ankyrin/band 3 ratios and in band 5 was observed. Conclusions Our data show that leukocyte-depleted RBCs present changes in the RBC morphology, membrane protein composition, enzymatic activity, and extracellular electrolyte concentration and pH. PMID:25960715

  20. Release of cytokines in stored whole blood and red cell concentrate: Effect of leukoreduction

    PubMed Central

    Shukla, Rinku; Patel, Tanvi; Gupte, Snehalata

    2015-01-01

    Background: Storage time of blood components plays a major role in the accumulation of cytokines causing adverse transfusion reactions. Aims: The aim was to study the trend in the levels of interleukin-6 (IL-6), IL-8, tumor necrosis factor-alpha (TNF-α) and regulated upon activation, normal T-cells expressed and secreted (RANTES) during storage of whole blood (WB) and red cell concentrate (RCC) and to study the effect of leukoreduction (LR). Materials and Methods: WB sample was taken on 0, 7, 14, 21, and between 28 and 35 days and plasma aliquots were frozen. Samples from RCC and buffy-coat depleted RCC prepared using Optipress II were collected on 0, 7, 14, 21 and between 28 and 35 days. Cytokine estimation was done using ELISA development kits. Normal range of cytokines was established using 0 day samples of WB. Statistical analysis was done using nonparametric tests. Results and Conclusion: The normal range of IL-6 was 0-23 pg/ml, IL-8 0-12 pg/ml, TNF-α 0-3 pg/ml, and RANTES 1200-2000 pg/ml. IL-6 was in normal range and showed a decreasing trend during storage. IL-8 levels increased significantly from 0 to 35 days. In RCC, the highest level was 480 pg/ml on 28th day. It was in the normal range in buffy-coat depleted RCC up to 28 days. RANTES level was significantly low in buffy-coat depleted RCC compared to RCC. We conclude that WB has high levels of IL-8 and RANTES. The levels of cytokines are affected by storage period and LR. Comparison of WB and buffy-coat depleted RCC shows significantly low levels of IL-6, IL-8, and RANTES in buffy-coat depleted RCC. This study emphasizes the use of red cell components instead of WB and buffy-coat depleted RCC instead of RCC. PMID:26420933

  1. Blood withdrawal affects iron store dynamics in primates with consequences on monoaminergic system function.

    PubMed

    Hyacinthe, C; De Deurwaerdere, P; Thiollier, T; Li, Q; Bezard, E; Ghorayeb, I

    2015-04-02

    Iron homeostasis is essential for the integrity of brain monoaminergic functions and its deregulation might be involved in neurological movement disorders such as the restless legs syndrome (RLS). Although iron metabolism breakdown concomitantly appears with monoaminergic system dysfunction in iron-deficient rodents and in RLS patients, the direct consequences of peripheral iron deficiency in the central nervous system (CNS) of non-human primates have received little attention. Here, we evaluated the peripheral iron-depletion impact on brain monoamine levels in macaque monkeys. After documenting circadian variations of iron and iron-related proteins (hemoglobin, ferritin and transferrin) in both serum and cerebrospinal fluid (CSF) of normal macaques, repeated blood withdrawals (RBW) were used to reduce peripheral iron-related parameter levels. Decreased serum iron levels were paradoxically associated with increased CSF iron concentrations. Despite limited consequences on tissue monoamine contents (dopamine - DA, 3, 4-dihydroxyphenylacetic acid - DOPAC, homovanillic acid, L-3, 4-dihydroxyphenylalanine - L-DOPA, 5-8 hydroxytryptamine - 5-HT, 5-hydroxyindoleacetic acid - 5-HIAA and noradrenaline) measured with post-mortem chromatography, we found distinct and region-dependent relationships of these tissue concentrations with CSF iron and/or serum iron and/or blood hemoglobin. Additionally, striatal extracellular DA, DOPAC and 5-HIAA levels evaluated by in vivo microdialysis showed a substantial increase, suggesting an overall increase in both DA and 5-HT tones. Finally, a trending increase in general locomotor activity, measured by actimetry, was observed in the most serum iron-depleted macaques. Taken together, our data are compatible with an increase in nigrostriatal DAergic function in the event of iron deficiency and point to a specific alteration of the 5-HT/DA interaction in the CNS that is possibly involved in the etiology of RLS.

  2. [Multivariate analysis of blood culture positive rate of ICU patients].

    PubMed

    Wu, A P; Liu, D; Chen, J; Li, X Y; Wang, H; An, Y Z

    2016-07-19

    To investigate the factors associated with positive results of blood culture and the impact of positive results on the prognosis of patients in ICU of Peking University People's Hospital. We retrospectively analyzed 1 008 blood culture results of 379 critical ill adult patients in ICU from July 1st, 2013 to June 30th, 2014. According to blood culture results, the patients were divided into positive and negative groups. The patients' maximal body temperature, sample collection times, number of bottles within 24 hours, routine hematological variables [(white blood cell count (WBC), percentage of neutrophils (NEU%), lymphocyte count (LYM), platelet count (PLT)], serum C-reactive protein (CRP), usage of antibiotics were compared between the two groups, as well as the patients' gender, age, duration of mechanical ventilation, length of ICU stay and hospital mortality rate. The total positiverate of blood culture of our study was 15.38%, and the positive rate of patients was 24.27%.When compared between positive group and negative group, the medians of sample collection times were 3 and 1(P<0.000 1); the medians of sample bottles were 4 and 4(P=0.001 2); the medians of WBC were 8.61×10(9)/L and 9.95×10(9)/L(P=0.001 7); and the medians of mechanical ventilation time were 179.5 hours and 47 hours(P<0.000 1); the medians of length of ICU stay were 17 days and 7 days(P<0.000 1), respectively. Hospital mortality rates in positive patients and negative patients were 35.87% and 20.21%(P=0.002 2), respectively. There was no significant difference(P>0.05) between the two groups in body temperature, NEU%, LYM, PLT, CRP or usage of antibiotics. Increasing the frequency of sampling and the bottles of blood culture will improve the positive rate of blood culture. The body temperature, WBC, NEU%, LYM, PLT, CRP, us age of antibiotics, gender and age have no effect on the positive rate of blood culture. The patients with positive blood culture results have longer duration of

  3. Factors affecting Brucella spp. blood cultures positivity in children.

    PubMed

    Apa, Hurşit; Devrim, Ilker; Memur, Seyma; Günay, Ilker; Gülfidan, Gamze; Celegen, Mehmet; Bayram, Nuri; Karaarslan, Utku; Bağ, Ozlem; Işgüder, Rana; Oztürk, Aysel; Inan, Seyhan; Unal, Nurrettin

    2013-03-01

    Brucella infections have a wide spectrum of symptoms especially in children, making the diagnosis a complicated process. The gold standard for the final diagnosis for brucellosis is to identify the Brucella spp. isolated from blood or bone marrow cultures. The main purpose of this work was to evaluate the factors affecting the isolation of Brucella spp. from blood cultures. In our study, the ratio of fever, presence of hepatomegaly, and splenomegaly were found to be higher in the bacteremic group. In addition, C-reactive protein levels and liver function enzymes were found to be higher in the bacteremic group. In our opinion, while evaluating the febrile child with suspected Brucella infection, we highly recommend sampling blood cultures regardless of the history of previous antimicrobial therapy and duration of the symptoms.

  4. Quantifying morphological heterogeneity: a study of more than 1,000,000 individual stored red blood cells

    PubMed Central

    Piety, Nathaniel Z.; Gifford, Sean C.; Yang, Xiaoxi; Shevkoplyas, Sergey S.

    2015-01-01

    Background and Objectives The morphology of red blood cells (RBCs) deteriorates progressively during hypothermic storage. The degree of deterioration varies between individual cells, resulting in a highly heterogeneous population of cells contained within each RBC unit. Current techniques capable of categorizing the morphology of individual stored RBCs are manual, laborious, error-prone procedures that limit the number of cells that can be studied. Our objective was to create a simple, automated system for high-throughput RBC morphology classification. Materials and Methods A simple microfluidic device, designed to enable rapid, consistent acquisition of images of optimally oriented RBCs, was fabricated using soft lithography. A custom image analysis algorithm was developed to categorize the morphology of each individual RBC in the acquired images. The system was used to determine morphology of individual RBCs in several RBC units stored hypothermically for 6–8 weeks. Results The system was used to automatically determine the distribution of cell diameter within each morphological class for >1,000,000 individual stored RBCs (speed: >10,000 cells/hour; accuracy: 91.9% low-resolution, 75.3% high-resolution). Diameter mean and standard deviation by morphology class: discocyte 7.80±0.49μm, echinocyte 1 7.61±0.63μm, echinocyte 2 7.02±0.61μm, echinocyte 3 6.47±0.42μm, sphero-echinocyte 6.01±0.26μm, spherocyte 6.02±0.27μm, stomatocyte 1 6.95±0.61μm, stomatocyte 2 7.32 ± 0.47μm. Conclusion The automated morphology classification procedure described in this study is significantly simpler, faster and less subjective than conventional manual procedures. The ability to evaluate the morphology of individual RBCs automatically, rapidly and in statistically significant numbers enabled us to perform the most extensive study of stored RBC morphology to date. PMID:25900518

  5. DNA quality and quantity from up to 16 years old post-mortem blood stored on FTA cards.

    PubMed

    Rahikainen, Anna-Liina; Palo, Jukka U; de Leeuw, Wiljo; Budowle, Bruce; Sajantila, Antti

    2016-04-01

    Blood samples preserved on FTA cards offer unique opportunities for genetic research. DNA recovered from these cards should be stable for long periods of time. However, it is not well established as how well the DNA stored on FTA card for substantial time periods meets the demands of forensic or genomic DNA analyses and especially so for from post-mortem (PM) samples in which the quality can vary upon initial collection. The aim of this study was to evaluate the time-dependent degradation on DNA quality and quantity extracted from up to 16 years old post-mortem bloodstained FTA cards. Four random FTA samples from eight time points spanning 1998 to 2013 (n=32) were collected and extracted in triplicate. The quantity and quality of the extracted DNA samples were determined with Quantifiler(®) Human Plus (HP) Quantification kit. Internal sample and sample-to-sample variation were evaluated by comparing recovered DNA yields. The DNA from the triplicate samplings were subsequently combined and normalized for further analysis. The practical effect of degradation on DNA quality was evaluated from normalized samples both with forensic and pharmacogenetic target markers. Our results suggest that (1) a PM change, e.g. blood clotting prior to sampling, affects the recovered DNA yield, creating both internal and sample-to-sample variation; (2) a negative correlation between the FTA card storage time and DNA quantity (r=-0.836 at the 0.01 level) was observed; (3) a positive correlation (r=0.738 at the level 0.01) was found between FTA card storage time and degradation levels. However, no inhibition was observed with the method used. The effect of degradation was manifested clearly with functional applications. Although complete STR-profiles were obtained for all samples, there was evidence of degradation manifested as decreased peak heights in the larger-sized amplicons. Lower amplification success was notable with the large 5.1 kb CYP2D6 gene fragment which strongly supports

  6. Assessment of anaerobic blood cultures in pediatric oncology patients.

    PubMed

    Monsonís Cabedo, Manuel; Rives Solá, Susana; Noguera-Julian, Antoni; Urrea Ayala, Mireia; Cruz Martinez, Ofelia; Gené Giralt, Amadeu

    2017-01-01

    The routine use of a single aerobic bottle for blood culture in pediatric patients has become commonplace, as anaerobic bacteria are not frequently involved in clinically significant infections. The aim of this study was to assess the usefulness of routinely performing anaerobic blood cultures in pediatric oncology patients. Prospective study was conducted on pediatric (<18 years) patients affected with febrile syndrome after receiving chemotherapy for hematological or solid malignancies. Samples were inoculated into pediatric aerobic and standard anaerobic bottles (BacT/Alert automatic system). Strains were considered clinically significant, or deemed as contaminants, depending on isolation circumstances and clinical criteria. A total of 876 blood cultures from 228 patients were processed during the 21-month study period (January 2014 to September 2015). Baseline diagnosis included 143 solid tumors and 67/18 cases of leukemia/lymphoma. Bacterial growth was detected in 90 (10.2%) blood cultures for 95 different isolates, of which 62 (7.1%)/63 isolates were considered clinically significant. Among the latter, 38 (60.3%) microorganisms grew in both aerobic and anaerobic bottles, 18 (28.6%) only in aerobic bottles, and 7 (11.1%) only in anaerobic bottles. Gram-negative bacilli (33; 52.4%), mainly from the Enterobacteriaceae family, were the most frequently isolated microorganisms. Overall, only 3 out of 90 isolates (3.3%) were strict anaerobes (Propionibacterium acnes), and all of them were deemed contaminants. Strict anaerobes did not cause significant infections in febrile pediatric oncology patients, and anaerobic blood culture bottles offered no additional advantages over aerobic media. Our results suggest that routine blood cultures should be solely processed in aerobic media in this group of patients. Copyright © 2016 Elsevier España, S.L.U. and Sociedad Española de Enfermedades Infecciosas y Microbiología Clínica. All rights reserved.

  7. Blood cultures in the evaluation of uncomplicated cellulitis.

    PubMed

    Bauer, Sophie; Aubert, Carole E; Richli, Mischa; Chuard, Christian

    2016-12-01

    The frequency of bacteremia and the array of microorganisms involved in cellulitis vary greatly among studies. Although current guidelines do not recommend routine blood culture in uncomplicated cellulitis, their implementation in clinical practice remains challenging. We therefore aimed to assess the frequency, determinants and microbiology of bacteremia in hospitalized patients with uncomplicated cellulitis. We retrospectively reviewed the medical records of all adult patients admitted at a primary-care hospital with a diagnosis of community-acquired uncomplicated cellulitis during a 4-year period. We looked at the factors associated with blood cultures sampling and at the incidence, determinants and microbiology of bacteremia in this population. Among the 476 patients hospitalized with a diagnosis of cellulitis, 250 (52.5%) had blood cultures. Fever, high C-reactive protein and lymphatic insufficiency were significantly associated with the sampling of blood cultures. Twelve (4.8%) patients had bacteremia. Alcoholism and duration of hospitalization were associated with bacteremia in multivariate analysis. Among the 12 patients with bacteremia, 9 had Streptococcus sp. and 3 had Staphylococcus aureus infection. In our study population with uncomplicated cellulitis, representative of unselected population admitted at primary-care hospitals, bacteremia was uncommon and not associated with discriminant patient characteristics, except for alcohol abuse. Episodes of bacteremia were exclusively due to gram-positive cocci susceptible to co-amoxicilin, a common first-line empirical therapy. In accordance with existing guidelines, we do not recommend to collect blood for cultures in uncomplicated cellulitis. Clinicians' awareness of guidelines and of the poor yield of blood cultures could reduce useless investigation. Copyright © 2016 European Federation of Internal Medicine. Published by Elsevier B.V. All rights reserved.

  8. Radiometric detection of yeasts in blood cultures of cancer patients

    SciTech Connect

    Hopfer, R.L.; Orengo, A.; Chesnut, S.; Wenglar, M.

    1980-09-01

    During a 12-month period, 19,457 blood cultures were collected. Yeasts were isolated from 193 cultures derived from 76 cancer patients. Candida albicans or Candida tropicalis accounted for 79% of isolates. Of the three methods compared, the radiometric method required 2.9 days to become positive, blind subculture required 2.6 days, and Gram stains required 1 day. However, the radiometric method was clearly superior in detecting positive cultures, since 73% of all cultures were first detected radiometrically, 22% were detected by subculture, and only 5% were detected by Gram stain. Although 93% of the isolates were detected by aerobic culture, five (7%) isolates were obtained only from anaerobic cultures. Seven days of incubation appear to be sufficient for the radiometric detection of yeasts.

  9. Storing red blood cells with vitamin C and N-acetylcysteine prevents oxidative stress-related lesions: a metabolomics overview

    PubMed Central

    Pallotta, Valeria; Gevi, Federica; D’Alessandro, Angelo; Zolla, Lello

    2014-01-01

    Background Recent advances in red blood cell metabolomics have paved the way for further improvements of storage solutions. Materials and methods In the present study, we exploited a validated high performance liquid chromatography-mass spectrometry analytical workflow to determine the effects of vitamin C and N-acetylcysteine supplementation (anti-oxidants) on the metabolome of erythrocytes stored in citrate-phosphate-dextrose saline-adenine-glucose-mannitol medium under blood bank conditions. Results We observed decreased energy metabolism fluxes (glycolysis and pentose phosphate pathway). A tentative explanation of this phenomenon could be related to the observed depression of the uptake of glucose, since glucose and ascorbate are known to compete for the same transporter. Anti-oxidant supplementation was effective in modulating the redox poise, through the promotion of glutathione homeostasis, which resulted in decreased haemolysis and less accumulation of malondialdehyde and oxidation by-products (including oxidized glutathione and prostaglandins). Discussion Anti-oxidants improved storage quality by coping with oxidative stress at the expense of glycolytic metabolism, although reservoirs of high energy phosphate compounds were preserved by reduced cyclic AMP-mediated release of ATP. PMID:25074788

  10. FM dyes enter via a store-operated calcium channel and modify calcium signaling of cultured astrocytes

    PubMed Central

    Li, Dongdong; Hérault, Karine; Oheim, Martin; Ropert, Nicole

    2009-01-01

    The amphiphilic fluorescent styryl pyridinium dyes FM1-43 and FM4-64 are used to probe activity-dependent synaptic vesicle cycling in neurons. Cultured astrocytes can internalize FM1-43 and FM4-64 inside vesicles but their uptake is insensitive to the elevation of cytosolic calcium (Ca2+) concentration and the underlying mechanism remains unclear. Here we used total internal reflection fluorescence microscopy and pharmacological tools to study the mechanisms of FM4-64 uptake into cultured astrocytes from mouse neocortex. Our data show that: (i) endocytosis is not a major route for FM4-64 uptake into astrocytes; (ii) FM4-64 enters astrocytes through an aqueous pore and strongly affects Ca2+ homeostasis; (iii) partitioning of FM4-64 into the outer leaflet of the plasma membrane results in a facilitation of store-operated Ca2+ entry (SOCE) channel gating; (iv) FM4-64 permeates and competes with Ca2+ for entry through a SOCE channel; (v) intracellular FM4-64 mobilizes Ca2+ from the endoplasmic reticulum stores, conveying a positive feedback to activate SOCE and to sustain dye uptake into astrocytes. Our study demonstrates that FM dyes are not markers of cycling vesicles in astrocytes and calls for a careful interpretation of FM fluorescence. PMID:20007370

  11. Quality and safety of red blood cells stored in two additive solutions subjected to multiple room temperature exposures.

    PubMed

    de Grandmont, M J; Ducas, E; Girard, M; Méthot, M; Brien, M; Thibault, L

    2014-10-01

    Many international standards state that red blood cell (RBC) products should be discarded if left out of controlled temperature storage for longer than 30 min to reduce the risk of bacterial growth and RBC loss of viability. This study aimed to verify whether repeated short-time exposures to room temperature (RT) influence RBCs quality and bacterial proliferation. Saline-adenine-glucose-mannitol (SAGM) and AS-3 RBC units were split and exposed to RT for 30 or 60 min on day 2, 7, 14, 21, and 42 of storage while reference units remained stored at 1-6°C. Red blood cell in vitro quality parameters were evaluated after each exposure. In a second experiment, SAGM and AS-3 RBC units were split and inoculated with Staphylococcus epidermidis (5 CFU/ml), Serratia marcescens (1 CFU/ml), and Serratia liquefaciens (1 CFU/ml). Reference units remained in storage while test units were exposed as described previously. Bacterial concentrations were investigated after each exposure. No differences were noticed between reference and test units in any of the in vitro parameters investigated. S. epidermidis did not grow in either reference or exposed RBCs. While S. marcescens did not grow in AS-3, bacterial growth was observed in RT-exposed SAGM RBCs on day 42. Similar growth was obtained for S. liquefaciens in the two additive solutions for both reference and test units. Short-time exposures to RT do not affect RBC quality and do not significantly influence bacterial growth. An expansion of the '30-minute' rule to 60 min should be considered by regulatory agencies. © 2014 International Society of Blood Transfusion.

  12. Innovation for reducing blood culture contamination: initial specimen diversion technique.

    PubMed

    Patton, Richard G; Schmitt, Timothy

    2010-12-01

    We hypothesized that diversion of the first milliliter of venipuncture blood-the initial specimen diversion technique (ISDT)-would eliminate incompletely sterilized fragments of skin from the culture specimen and significantly reduce our blood culture contamination rate (R). We studied our hypothesis prospectively beginning with our control culture (C) definition: one venipuncture with two sequentially obtained specimens, 10 ml each, the first specimen (M1) for aerobic and the second (M2) for anaerobic media. The test ISDT culture (D) was identical, with the exception that each was preceded by diverting a 1-ml sample (DS) from the same venipuncture. During the first of two sequential 9-month periods, we captured D versus C data (n=3,733), where DMXR and CMXR are R for D and C specimens. Our hypothesis predicted DS would divert soiled skin fragments from DM1, and therefore, CM1R would be significantly greater than DM1R. This was confirmed by CM1R (30/1,061 [2.8%]) less DM1R (37/2,672 [1.4%]; P=0.005), which equals 1.4%. For the second 9-month follow-up period, data were compiled for all cultures (n=4,143), where ADMXR is R for all (A) diversion specimens, enabling comparison to test ISDT. Our hypothesis predicted no significant differences for test ISDT versus all ISDT. This was confirmed by DM1R (37/2,672 [1.4%]) versus ADM1R (42/4,143 [1.0%]; P=0.17) and DM2R (21/2,672 [0.80%]) versus ADM2R (39/4,143 [0.94%]; P=0.50). We conclude that our hypothesis is valid: venipuncture needles soil blood culture specimens with unsterilized skin fragments and increase R, and ISDT significantly reduces R from venipuncture-obtained blood culture specimens.

  13. Improved blood culture identification by FilmArray in cultures from regional hospitals compared with teaching hospital cultures.

    PubMed

    Inglis, Timothy J J; Bzdyl, Nicole; Chua, I-Ly Joanna; Urosevic, Nadezda M; Leung, Michael J; Geelhoed, Elizabeth

    2016-01-01

    Rapid identification of bacteria isolated from blood cultures by direct matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) is now in wide spread use in major centres but is not yet feasible in smaller hospital laboratories. A FilmArray multiplex PCR panel for blood culture isolate identification (BCID) provides an alternative approach to near point-of-care microbial identification in regional hospitals. We assessed the accuracy and time to identification of the BCID FilmArray in a consecutive series of 149 blood cultures from 143 patients in a teaching hospital and smaller regional hospitals, currently identified by direct MALDI-TOF and proprietary molecular methods. The BCID FilmArray contained 18 of 34 species and 20 of 23 species isolated from teaching and regional hospital, respectively. Overall, 85 % of the teaching hospital and 100 % of the regional hospital monomicrobial blood cultures were identified, compared with 60 and 68 %, respectively, for direct MALDI-TOF on the same cultures. There were no incorrect results from blood cultures containing Staphylococcus aureus, streptococci, Pseudomonas aeruginosa or Enterobacteriaceae. The three discrepant results were all in mixed cultures. The mean reduction in time to identification of blood culture isolates was 53 h, which did not include the time required to transport cultures from regional centres to a central laboratory. The overall performance of the BCID FilmArray is stronger in blood cultures from smaller regional hospitals that encounter a narrower range of bacterial species dominated by the commonest species. This approach is more suited to smaller clinical laboratories than the MALDI-TOF direct method.

  14. Clinical significance of Bacillus species isolated from blood cultures.

    PubMed

    Weber, D J; Saviteer, S M; Rutala, W A; Thomann, C A

    1989-06-01

    To determine the clinical significance of blood isolates of Bacillus, we reviewed all blood cultures obtained at North Carolina Memorial Hospital between 1981 and 1985. Over the five-year study period the number of patients (incidence per 10,000 hospital admissions) from whom Bacillus was isolated increased from 4.97 in 1981 to 12.5 in 1985. The incidence per 1,000 blood cultures also increased from 1.12 in 1981 to 2.33 in 1985. Review of the medical records of 78 of the 95 patients (82%) with positive cultures allowed retrospective classification of five isolates (6.4%) as clinically significant, 33 isolates (42.3%) as possibly significant, and 40 isolates (51.3%) as nonsignificant. Underlying diseases in patients with clinically significant Bacillus bacteremia included burn trauma in two, leukemia in one, carcinoma in one, and gastrointestinal hemorrhage in one. All isolates judged to be clinically significant and the majority of possibly significant isolates were B cereus. We conclude that the isolation of Bacillus species from blood cultures is clinically significant in 5% to 10% of cases, that the incidence of Bacillus bacteremia is increasing, and that burn trauma should be added to the list of conditions known to predispose to clinically significant Bacillus bacteremia.

  15. Controlled clinical comparison of three commercial blood culture systems.

    PubMed

    Frank, U; Malkotsis, D; Mlangeni, D; Daschner, F D

    1999-04-01

    In a controlled clinical comparison, three commercial blood culture systems--the standard aerobic BacT/Alert bottle (STD), the aerobic BacT/Alert FAN bottle (FAN) and the Isolator system (ISO; Wampole Laboratories, USA) were compared for their ability to detect aerobic and facultatively anaerobic microorganisms. A total of 945 BacT/Alert (STD and FAN) blood culture sets were compared. Of these, 110 blood culture sets (11.6%) yielded growth of 116 clinically significant bacterial and fungal isolates. Microorganisms were recovered from 10.7% (101/945) of the FAN bottles compared to 8.9% (84/945) of the STD bottles. Of the significant isolates, 78 (67.2%) were recovered by both bottles, 29 (25%) by the FAN bottle only and nine (7.8%) by the STD bottle only (P<0.01). Along with 56.1% (530/945) of BacT/Alert blood culture sets, a concomitant ISO tube was obtained. Of the triple (STD + FAN + ISO) blood culture sets, 54 (10.2%) yielded growth of 59 clinically relevant isolates. Microorganisms were detected in 9.1% (48/530) of the FAN bottles, 8.3% (44/530) of the STD bottles and 4% (21/530) of the ISO tubes (P<0.001). Overall, the BacT/Alert system detected more clinically significant microorganisms than the ISO tube; the STD and the FAN bottle each recovered significantly more staphylococci (P<0.01 and P<0.001, respectively) and gram-negative rods (P<0.01, both). In conclusion, the BacT/Alert FAN bottle performed better than the BacT/Alert STD bottle; both BacT/Alert bottles, however, were superior to the ISO tube in terms of recovery of clinically significant microorganisms, including gram-positive and gram-negative bacteria.

  16. Haemostasis monitored in stored red blood cells, plasma and platelet concentrates in the proportion of 4 :  4 :  1 diluted with crystalloids and colloids.

    PubMed

    Ågren, Anna; Edgren, Gustaf; Ambrosio, Daniela; Gryfelt, Gunilla; Östlund, Anders; Wikman, Agneta

    2016-04-01

    The aim of this in-vitro study was to evaluate haemostasis analysed with thromboelastometry and blood gas and blood count variables, in stored blood components and the effects after dilution with Ringer[Combining Acute Accent]s acetate, albumin and hydroxyethyl starch (HES). Aliquots from stored red blood cells, plasma and platelet concentrates were mixed in the proportion of 4 : 4 : 1 and analysed with rotational thromboelastometry (ROTEM), blood count [haemoglobin (Hb), haematocrit, platelet count] and blood gas (pH, calcium, sodium, potassium, glucose levels). The blood mix was thereafter diluted 20 and 33% with Ringer's acetate, albumin or HES. The stored blood component mix in a ratio of 4 : 4 : 1 had a low pH (7.11 ± 0.03, mean ± standard deviation), nonmeasurable calcium level, and high concentrations of sodium, potassium and glucose but ROTEM curves within normal range after recalcification. With Ringer's acetate dilution, the ROTEM variables changed almost linearly with increasing dilution volume. When albumin was used in the 33% dilution, the clot firmness of the fibrin clot (FibTEM) was further reduced, and with HES dilution, there was a pronounced impairment. The stored blood mix had a low pH and calcium level, both of which might have a significant influence on the coagulation process but normal ROTEM curves after recalcification. Dilution with Ringer's acetate and albumin resulted in moderate deterioration, while dilution with HES showed severely impaired haemostasis.

  17. Prefibrillar transthyretin oligomers and cold stored native tetrameric transthyretin are cytotoxic in cell culture

    SciTech Connect

    Soergjerd, Karin; Klingstedt, Therese; Lindgren, Mikael; Kagedal, Katarina; Hammarstroem, Per

    2008-12-26

    Recent studies suggest that soluble, oligomeric species, which are intermediates in the fibril formation process in amyloid disease, might be the key species in amyloid pathogenesis. Soluble oligomers of human wild type transthyretin (TTR) were produced to elucidate oligomer properties. Employing ThT fluorescence, time-resolved fluorescence anisotropy of pyrene-labeled TTR, chemical cross-linking, and electron microscopy we demonstrated that early formed soluble oligomers (within minutes) from A-state TTR comprised on the average 20-30 TTR monomers. When administered to neuroblastoma cells these early oligomers proved highly cytotoxic and induced apoptosis after 48 h of incubation. More mature fibrils (>24 h of fibrillation) were non-toxic. Surprisingly, we also found that native tetrameric TTR, when purified and stored under cold conditions (4 deg. C) was highly cytotoxic. The effect could be partially restored by increasing the temperature of the protein. The cytotoxic effects of native tetrameric TTR likely stems from a hitherto unexplored low temperature induced rearrangement of the tetramer conformation that possibly is related to the conformation of misfolded TTR in amyloigogenic oligomers.

  18. Red blood cell distribution width is not a reliable biomarker for low iron stores in children with cystic fibrosis.

    PubMed

    Akkermans, M D; Uijterschout, L; Nuijsink, M; Hendriks, D M; van Goudoever, J B; Brus, F

    2017-02-01

    Low iron stores in children, absolute iron deficiency (AID), can lead to impaired neurodevelopment and requires iron therapy. In the presence of infection/inflammation, like in cystic fibrosis (CF), serum ferritin (SF) is not a reliable biomarker for AID. Red blood cell distribution width (RDW) is a promising alternative reported not to be influenced by infection in healthy children. Currently, there are no data on the diagnostic capacity of RDW to detect AID in pediatric CF patients. This was a prospective observational study that investigated iron status biomarkers in 53 Dutch pediatric CF patients. AID was defined using World Health Organization criteria for SF in stable patients (no recent pulmonary exacerbation) and C-reactive protein (CRP) ≤10 mg/l. Patients with AID had higher RDW levels than patients without AID (p = 0.019). An RDW ≥13.2% showed the following test statistics: sensitivity 100%; specificity 39.4%; positive predictive value 20%; and negative predictive value 100%. Furthermore, we found a correlation between RDW and CRP in the total group that originated from the stable patients (r = 0.308; p = 0.042). In conclusion, the diagnostic capacity of RDW for detecting AID in pediatric CF patients seems limited because RDW levels might also be influenced by chronic infection/inflammation in these patients.

  19. Stored red blood cell susceptibility to in vitro transfusion-associated stress conditions is higher after longer storage and increased by storage in saline-adenine-glucose-mannitol compared to AS-1.

    PubMed

    Mittag, Diana; Sran, Amrita; Chan, Kasey S; Boland, Martin P; Bandala-Sanchez, Esther; Huet, Olivier; Xu, William; Sparrow, Rosemary L

    2015-09-01

    Biochemical changes induced in red blood cells (RBCs) during storage may impair their function upon transfusion. Transfusion-associated stresses may further amplify storage lesion effects including increased phosphatidylserine (PS) exposure at the RBC membrane, microparticle (MP) release, and adhesion to endothelial cells (ECs). RBC stress susceptibility in vitro was investigated in relation to storage time and additive solution. Leukoreduced whole blood donations (n = 18) were paired, mixed, and resplit before separating the RBCs for storage in saline-adenine-glucose-mannitol (SAGM) or AS-1. Samples were taken after 3, 21, or 35 days. For oxidative stress treatment, RBCs were exposed to 0.5 mmol/L tert-butylhydroperoxide. Transfusion-associated stress was simulated by overnight culture at 37 °C with plasma containing inflammatory mediators. PS exposure and MPs were measured by flow cytometry and adhesion to ECs was tested under flow conditions. PS specificity of adhesion was tested by blocking with PS-containing lipid vesicles. Oxidative stress induced significantly higher PS exposure and adhesion to ECs in RBCs stored for 35 days compared to 3 days (p < 0.04). PS-containing vesicles blocked RBC-EC adhesion. After overnight culture with or without plasma, PS exposure and EC adhesion were significantly increased (p < 0.05). MP numbers increased with longer RBC storage and after RBC culture with plasma. Culture conditions influenced MP numbers from Day 35 RBCs. RBCs stored in SAGM had significantly higher PS exposure after stress treatment than AS-1 RBCs (p < 0.02). Storage for 35 days significantly increased RBC susceptibility to oxidative and in vitro transfusion-associated stresses and was higher for RBCs stored in SAGM compared to AS-1. © 2015 AABB.

  20. Stored red blood cell susceptibility to in vitro transfusion-associated stress conditions is higher after longer storage and increased by storage in saline-adenine-glucose-mannitol (SAGM) compared to AS-1 solution

    PubMed Central

    Mittag, Diana; Sran, Amrita; Chan, Kasey S; Boland, Martin P; Bandala-Sanchez, Esther; Huet, Olivier; Xu, William; Sparrow, Rosemary L

    2015-01-01

    Background Biochemical changes induced in red blood cells (RBCs) during storage may impair their function upon transfusion. Transfusion-associated stresses may further amplify storage lesion effects including increased phosphatidylserine (PS)-exposure at the RBC membrane, microparticle (MP)-release and adhesion to endothelial cells (ECs). RBC stress susceptibility in vitro was investigated in relation to storage time and additive solution. Study design and methods Leukocyte-reduced whole blood donations (n=18) were paired, mixed and re-split before separating the RBCs for storage in SAGM or AS-1 solutions. Samples were taken after 3, 21 or 35 days. For oxidative stress treatment RBCs were exposed to 0.5mM tert-butylhydroperoxide. Transfusion-associated stress was simulated by overnight culture at 37 °C with plasma containing inflammatory mediators. PS-exposure and MPs were measured by flow cytometry and adhesion to EC was tested under flow conditions. PS specificity of adhesion was tested by blocking with PS-containing lipid vesicles. Results Oxidative stress induced significantly higher PS-exposure and adhesion to ECs in RBCs stored for 35 days compared to 3 days (p<0.04). PS-containing vesicles blocked RBC-EC adhesion. After overnight culture with or without plasma, PS-exposure and EC-adhesion were significantly increased (p<0.05). MP numbers increased with longer RBC storage and after RBC culture with plasma. Culture conditions influenced MP numbers from day 35 RBCs. RBCs stored in SAGM had significantly higher PS-exposure after stress treatment than AS-1 RBCs (p<0.02). Conclusion Storage for 35 days significantly increased RBC susceptibility to oxidative and in vitro transfusion-associated stresses and was higher for RBCs stored in SAGM compared to AS-1. PMID:25968419

  1. Quantitation of Candida CFU in initial positive blood cultures.

    PubMed

    Pfeiffer, Christopher D; Samsa, Gregory P; Schell, Wiley A; Reller, L Barth; Perfect, John R; Alexander, Barbara D

    2011-08-01

    One potential limitation of DNA-based molecular diagnostic tests for Candida bloodstream infection (BSI) is organism burden, which is not sufficiently characterized. We hypothesized that the number of CFU per milliliter (CFU/ml) present in an episode of Candida BSI is too low for reliable DNA-based diagnostics. In this study, we determined Candida burden in the first positive blood culture and explored factors that affect organism numbers and patient outcomes. We reviewed records of consecutive patients with a positive blood culture for Candida in the lysis-centrifugation blood culture system (Isolator, Wampole Laboratories, Cranbury, NJ) from 1987 to 1991. Descriptive statistics and logistic regression analyses were performed. One hundred fifty-two episodes of Candida BSI were analyzed. Patient characteristics included adult age (72%), indwelling central venous catheters (83%), recent surgery (29%), neutropenia (24%), transplant (14%), and other immune suppression (21%). Rates of treatment success and 30-day mortality for candidemia were each 51%. The median CFU/ml was 1 (mode 0.1, range 0.1 to >1,000). In the multivariate analysis, pediatric patients were more likely than adults to have high organism burdens (odds ratio [OR], 10.7; 95% confidence interval [95% CI], 4.3 to 26.5). Initial organism density did not affect patient outcome. Candida CFU/ml in the first positive blood culture of a BSI episode varies greatly; >50% of cultures had ≤1 CFU/ml, a concentration below the experimental yeast cell threshold for reliable DNA-based diagnostics. DNA-based diagnostics for Candida BSI will be challenged by low organism density and the need for sufficient specimen volume; future research on alternate targets is warranted.

  2. Addition of Sodium Pyruvate to Stored Red Blood Cells Attenuates Liver Injury in a Murine Transfusion Model

    PubMed Central

    2016-01-01

    RBCs undergo numerous changes during storage and stored RBCs may induce adverse effects, ultimately resulting in organ injury in transfusion recipients. We tested the hypothesis that the addition of SP to stored RBCs would improve the quality of the stored RBCs and mitigate liver injury after transfusion in a murine model. RBCs were harvested from C57BL/6J mice and stored for 14 days in CPDA-1 containing either a solution of SP in saline or saline alone. Haemolysis, the 24-hour posttransfusion recovery, the oxygen-carrying capacity, and the SOD activity of stored RBCs were evaluated. The plasma biochemistry, hepatic MDA level, MPO activity, IL-6, TNF-α concentrations, and histopathology were measured two hours after the transfusion of stored RBCs. Compared with RBCs stored in CPDA-1 and saline, the addition of SP to stored RBCs restored their oxygen-carrying capacity and SOD activity, reduced the AST activity, BUN concentrations, and LDH activity in the plasma, and decreased the MDA level, MPO activity, and concentrations of IL-6 and TNF-α in the liver. These data indicate that the addition of SP to RBCs during storage has a beneficial effect on storage lesions in vitro and subsequently alleviates liver injury after the transfusion of stored RBCs in vivo. PMID:27746589

  3. The in vitro and in vivo evaluation of whole blood and red cell concentrates drawn on CPDA-1 and stored in a non-DEHP plasticized PVC container.

    PubMed

    Seidl, S; Gosda, W; Reppucci, A J

    1991-01-01

    A new polyvinyl chloride (PVC) blood storage container, PL 2209, plasticized with butyryl-n-trihexyl-citrate (BTHC) instead of diethylhexyl phthalate (DEHP) was used for in vitro and in vivo studies. Whole blood and red cell concentrates drawn on CPDA-1 were stored 42 days at 1-6 degrees C. Various biochemical parameters were tested and compared to those values seen in the literature for products stored in DEHP-plasticized containers. Measurements of lactate production, glucose consumption, sodium and potassium ions for both whole blood and red cells were compatible with published data. Adenosine triphosphate (ATP) values were on the average 73% for the red cell units and 80% for the whole blood units after 35 days storage. Hemolysis was 0.43% for the red cell concentrate and 0.38% for the whole blood units after the same storage period. Autologous recovery studies after 35 days of storage for both the whole blood units and red cell concentrate gave 80% recovery 24 h after reinjection. From these data it can be concluded that PL 2209 BTHC-plasticized PVC blood storage containers allow 35 days storage of blood draw on CPDA-1 and can be considered as an alternative to DEHP-plasticized PVC containers.

  4. First Report of Clostridium lavalense Isolated in Human Blood Cultures.

    PubMed

    Garceau, Richard; Bourque, Christine; Thibault, Louise; Côté, Jean-Charles; Longtin, Jean; Domingo, Marc-Christian

    2016-01-01

    An 88-year-old man was admitted to the hospital with worsening malaise, fever, and weakness. Anaerobic blood culture bottles revealed the presence of an anaerobic, Gram-positive sporulated bacillus. Empirical antibiotherapy with intravenous piperacillin-tazobactam was initiated. The patient defervesced after four days and was switched to oral amoxicillin on his 6th day of antibiotic therapy and later discharged from the hospital. Four months later, he had recovered. The bacterium was initially identified as Clostridium butyricum using anaerobic manual identification panel. 16S rRNA gene sequence and phylogenetic analysis showed the bacterium to be Clostridium lavalense, a recently described species with no previously published case of isolation in human diagnostic samples so far. This is the first report of Clostridium lavalense isolation from human blood cultures. Further studies are needed in order to elucidate the role of Clostridium lavalense in human disease and its virulence factors.

  5. First Report of Clostridium lavalense Isolated in Human Blood Cultures

    PubMed Central

    Bourque, Christine; Thibault, Louise; Côté, Jean-Charles; Domingo, Marc-Christian

    2016-01-01

    An 88-year-old man was admitted to the hospital with worsening malaise, fever, and weakness. Anaerobic blood culture bottles revealed the presence of an anaerobic, Gram-positive sporulated bacillus. Empirical antibiotherapy with intravenous piperacillin-tazobactam was initiated. The patient defervesced after four days and was switched to oral amoxicillin on his 6th day of antibiotic therapy and later discharged from the hospital. Four months later, he had recovered. The bacterium was initially identified as Clostridium butyricum using anaerobic manual identification panel. 16S rRNA gene sequence and phylogenetic analysis showed the bacterium to be Clostridium lavalense, a recently described species with no previously published case of isolation in human diagnostic samples so far. This is the first report of Clostridium lavalense isolation from human blood cultures. Further studies are needed in order to elucidate the role of Clostridium lavalense in human disease and its virulence factors. PMID:27478446

  6. Rapid Detection of ESBL-Producing Enterobacteriaceae in Blood Cultures

    PubMed Central

    Dortet, Laurent; Poirel, Laurent

    2015-01-01

    We rapidly identified extended-spectrum β-lactamase (ESBL) producers prospectively among 245 gram-negative bacilli–positive cultured blood specimens using the Rapid ESBL Nordmann/Dortet/Poirel test and direct bacterial identification using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry. This combination identified ESBL-producing Enterobacteriaceae within 30 min and had high predictive values. PMID:25695535

  7. Enterococcus spp. in a single blood culture: bacteremia or contamination?

    PubMed

    Khatib, R; Labalo, V; Sharma, M; Johnson, L B; Riederer, K

    2017-03-01

    We retrospectively evaluated adult cases with Enterococcus spp. in 1 blood culture (BC) (1/1/2010-12/31/2015; n=294) and stratified them into bacteremia or contamination. Contamination frequency was similar in community versus hospital-onset, E. faecalis versus E. faecium, and number of BC drawn per day. Contamination predictors were vancomycin-resistance, ampicillin-resistance, commensal organism copresence, and nonurinary/abdominal sources.

  8. [Identification and antimicrobial susceptibility testing of positive blood culture isolates from briefly incubated solid medium cultures].

    PubMed

    Ballestero-Téllez, Mónica; Recacha, Esther; de Cueto, Marina; Pascual, Álvaro

    2016-02-16

    Mass spectrometry Matrix-Assisted Laser Desorption-Ionization Time-of-Flight (MALDI-TOF) helps in the rapid identification of microorganisms causing blood stream infection. Rapid and reliable methods are required to decrease the turnaround time for reporting antimicrobial susceptibility results from blood culture isolates. An evaluation was performed on the reliability of a method for antimicrobial susceptibility testing of positive blood culture isolates from briefly incubated solid medium cultures. The agreement between the evaluated and standard methods was 99.3%. The major and minor error rates were 0.4% and 0.3%, respectively, and no very major errors were observed. The inoculation of briefly incubated solid medium cultures into antimicrobial susceptibility testing panels is an easy and reliable technique, and helps to decrease the turnaround time for reporting antimicrobial susceptibility results of positive blood cultures. Copyright © 2016 Elsevier España, S.L.U. y Sociedad Española de Enfermedades Infecciosas y Microbiología Clínica. All rights reserved.

  9. Measurement of HbA1c from stored whole blood samples in the Atherosclerosis Risk in Communities study

    PubMed Central

    SELVIN, Elizabeth; CORESH, Josef; ZHU, Hong; FOLSOM, Aaron; STEFFES, Michael W.

    2010-01-01

    Background The aims of the present study were to demonstrate the reliability of HbA1c measurements across two time periods and to compare these measurements with HbA1c distribution in the general US population. Methods HbA1c was measured in 14 069 whole blood samples in the Atherosclerosis Risk in Communities (ARIC) study using different HPLC instruments during two time periods, namely 2003–2004 and 2007–2008. At the time of measurement, samples had been in storage at –70°C for 14–18 years. To assess differences in values, HbA1c measurements were repeated in 383 samples at both periods. Indirect comparisons were made by comparing our measurements against those from a nationally representative study. Results The coefficients of variation for quality control samples were 1.8% (n = 89) in 2003–2004 and 1.4% (n = 259) in 2007–2008. The correlation between measurements at the two time points was high (r = 0.99), but with a slight bias: 0.29% points higher in 2007–2008 versus 2003–2004 (n = 383; P < 0.0001). The comparison yielded the following Deming regression equation: y(2007–2008) = 0.073+1.034x(2003–2004). After alignment using this equation, the distribution of HbA1c in the ARIC study was similar to that in the national study using fresh samples. Conclusions Measurements of HbA1c from samples stored for 14–18 years are highly reliable when using state-of-the-art HPLC instruments, but with some bias introduced over time. The HbA1c data now available in the ARIC study should be invaluable for investigations into the clinical utility of HbA1c as a diagnostic test for diabetes. PMID:20923494

  10. Measurement of HbA1c from stored whole blood samples in the Atherosclerosis Risk in Communities study.

    PubMed

    Selvin, Elizabeth; Coresh, Josef; Zhu, Hong; Folsom, Aaron; Steffes, Michael W

    2010-06-01

    The aims of the present study were to demonstrate the reliability of HbA1c measurements during two time periods and to compare these measurements with HbA1c distribution in the general US population. HbA1c was measured in 14,069 whole blood samples in the Atherosclerosis Risk in Communities (ARIC) study using different HPLC instruments across two time periods, namely 2003-2004 and 2007-2008. At the time of measurement, samples had been in storage at -70°C for up to 18 years. To assess differences in values, HbA1c measurements were repeated in 383 samples at both periods. Indirect comparisons were made by comparing our measurements against those from a nationally representative study. The coefficients of variation for quality control samples were 1.8% (n = 89) in 2003-2004 and 1.4% (n = 259) in 2007-2008. The correlation between measurements at the two time points was high (r = 0.99), but with a slight bias: 0.29% points higher in 2007-2008 vs 2003-2004 (n = 383; P < 0.0001). The comparison yielded the following Deming regression equation: y((2007-2008)) = 0.073 + 1.034x((2003-2004)) . After alignment using this equation, the distribution of HbA1c in the ARIC study was similar to that in the national study using fresh samples. Measurements of HbA1c from samples stored for up to 18 years are highly reliable when using state-of-the-art HPLC instruments, but with some bias introduced over time. The HbA1c data now available in the ARIC study should be invaluable for investigations into the clinical utility of HbA1c as a diagnostic test for diabetes. © 2010 Ruijin Hospital, Shanghai Jiaotong University School of Medicine and Blackwell Publishing Asia Pty Ltd.

  11. Decreased mortality associated with prompt Gram staining of blood cultures.

    PubMed

    Barenfanger, Joan; Graham, Donald R; Kolluri, Lavanya; Sangwan, Gaurav; Lawhorn, Jerry; Drake, Cheryl A; Verhulst, Steven J; Peterson, Ryan; Moja, Lauren B; Ertmoed, Matthew M; Moja, Ashley B; Shevlin, Douglas W; Vautrain, Robert; Callahan, Charles D

    2008-12-01

    Gram stains of positive blood cultures are the most important factor influencing appropriate therapy. The sooner appropriate therapy is initiated, the better. Therefore, it is reasonable to expect that the sooner Gram stains are performed, the better. To determine the value of timely Gram stains and whether improvement in Gram stain turnaround time (TAT) is feasible, we compared data for matched pairs of patients with cultures processed promptly (<1 hour TAT) with data for patients with cultures not processed promptly (> or =1 hour TAT) and then monitored TAT by control charting.In 99 matched pairs, average difference in time to detection of positive blood cultures within a pair of patients was less than 0.1 hour. For the less than 1 hour TAT group, the average TAT and crude mortality were 0.1 hour and 10.1%, respectively; for the 1 hour or longer TAT group, they were 3.3 hours and 19.2%, respectively (P < .0001 and P = .0389, respectively). After multifaceted efforts, we achieved significant improvement in the TAT for Gram stains.

  12. Corneo-scleral rim cultures: donor contamination a case of fungal endophthalmitis transmitted by K-Sol stored cornea.

    PubMed

    Fong, L P; Gladstone, D; Casey, T A

    1988-01-01

    This retrospective study of 549 corneo-scleral rim cultures shows that gentamicin, used in MK and K-Sol medium storage at 4 degrees C, has decreased donor contamination from 43% in whole-globe storage to 13%, but failed to eliminate coagulase negative staphylococci (37%), streptococci (28%) and fungi (28%). Donor-to-host transmitted staphylococcal and streptococcal endophthalmitis have been reported previously. We present the first documented case of donor-to-recipient transmitted fungal endophthalmitis following corneal transplantation using corneas stored in MK or K-Sol solution at 4 degrees C; Candida albicans was isolated. Recommendations are made to assess critically the true incidence of donor fungal contamination and the necessity of adding anti-mycotic agents to preservation medium for 4 degrees C storage. In the absence of ideal antimicrobial cover for corneal preservation solutions, stringent prophylactic measures to reduce contamination and continued monitoring of corneo-scleral rim cultures are warranted, if the poor visual consequences of donor-to-host transmitted endophthalmitis are to be avoided.

  13. Coagulation Function of Stored Whole Blood is Preserved for 14 Days in Austere Conditions: A ROTEM Feasibility Study During a Norwegian Antipiracy Mission and Comparison to Equal Ratio Reconstituted Blood

    DTIC Science & Technology

    2015-06-24

    hulled inflatable boats, during which freeze-dried plasma and leukoreduced, group O low anti-A/ anti-B titer, cold -stored whole blood were stored in...components, we concluded that the only feasible option open to the prehospital team was the use of freeze-dried plasma and cold -stored leukoreduced whole...protocol, combat medics took Golden Hour Boxes packed with 2 U of freeze-dried plasma and 2 U of leukore- duced, group O low anti-A/anti-B titer, cold

  14. THE IN VITRO TRANSFORMATION OF FROZEN-STORED LYMPHOCYTES IN THE MIXED LYMPHOCYTE REACTION AND IN CULTURE WITH PHYTOHEMAGGLUTININ AND SPECIFIC ANTIGENS

    PubMed Central

    Mangi, Richard J.; Mardiney, Michael R.

    1970-01-01

    We have investigated the in vitro transformation of lymphocytes that have been frozen and stored at low temperatures. Cells frozen at different times and stored for various times do transform in a reproducible manner when placed in culture with PHA or as one population of the two-way or the one-way MLR. When frozen-stored lymphocytes are cultured with specific antigen or as both partners in the MLR the response is minimal. The freezing process destroys neutrophils, and the remaining population transforms in a manner similar to cultures of purified lymphocytes. Practical aspects of the technique are discussed and we have suggested possible practical applications for future investigation. PMID:5523965

  15. Multicenter evaluation of the Verigene Gram-negative blood culture nucleic acid test for rapid detection of bacteria and resistance determinants in positive blood cultures.

    PubMed

    Uno, Naoki; Suzuki, Hiromichi; Yamakawa, Hiromi; Yamada, Maiko; Yaguchi, Yuji; Notake, Shigeyuki; Tamai, Kiyoko; Yanagisawa, Hideji; Misawa, Shigeki; Yanagihara, Katsunori

    2015-12-01

    The Verigene Gram-Negative Blood Culture Nucleic Acid Test (BC-GN) is a microarray-based assay that enables rapid detection of 9 common Gram-negative bacteria and 6 resistance determinants directly from positive blood cultures. We compared the performance of BC-GN with currently used automated systems, testing 141 clinical blood cultures and 205 spiked blood cultures. For identification of BC-GN target organisms in clinical and spiked blood cultures, the BC-GN assay showed 98.5% (130/132) and 98.9% (182/184) concordance, respectively. Of 140 resistance genes positively detected in clinical and spiked blood cultures with the BC-GN test, 139 (99.3%) were confirmed by PCR, and the detection results were consistent with the resistance phenotypes observed. The BC-GN assay, thus, can potentially improve care for sepsis patients by enabling timely detection and targeted antimicrobial therapy. Copyright © 2015 Elsevier Inc. All rights reserved.

  16. Improved Diagnosis of Prosthetic Joint Infection by Culturing Periprosthetic Tissue Specimens in Blood Culture Bottles

    PubMed Central

    Peel, Trisha N.; Dylla, Brenda L.; Hughes, John G.; Lynch, David T.; Greenwood-Quaintance, Kerryl E.; Cheng, Allen C.; Mandrekar, Jayawant N.

    2016-01-01

    ABSTRACT Despite known low sensitivity, culture of periprosthetic tissue specimens on agars and in broths is routine. Culture of periprosthetic tissue samples in blood culture bottles (BCBs) is potentially more convenient, but it has been evaluated in a limited way and has not been widely adopted. The aim of this study was to compare the sensitivity and specificity of inoculation of periprosthetic tissue specimens into blood culture bottles with standard agar and thioglycolate broth culture, applying Bayesian latent class modeling (LCM) in addition to applying the Infectious Diseases Society of America (IDSA) criteria for prosthetic joint infection. This prospective cohort study was conducted over a 9-month period (August 2013 to April 2014) at the Mayo Clinic, Rochester, MN, and included all consecutive patients undergoing revision arthroplasty. Overall, 369 subjects were studied; 117 (32%) met IDSA criteria for prosthetic joint infection, and 82% had late chronic infection. Applying LCM, inoculation of tissues into BCBs was associated with a 47% improvement in sensitivity compared to the sensitivity of conventional agar and broth cultures (92.1 versus 62.6%, respectively); this magnitude of change was similar when IDSA criteria were applied (60.7 versus 44.4%, respectively; P = 0.003). The time to microorganism detection was shorter with BCBs than with standard media (P < 0.0001), with aerobic and anaerobic BCBs yielding positive results within a median of 21 and 23 h, respectively. Results of our study demonstrate that the semiautomated method of periprosthetic tissue culture in blood culture bottles is more sensitive than and as specific as agar and thioglycolate broth cultures and yields results faster. PMID:26733067

  17. Development of subsequent bloodstream infection in patients with positive Hickman catheter blood cultures and negative peripheral blood cultures.

    PubMed

    Park, Ki-Ho; Cho, Oh-Hyun; Lee, Sang-Oh; Choi, Sang-Ho; Kim, Yang Soo; Woo, Jun Hee; Kim, Mi-Na; Kim, Dae-Young; Lee, Jung-Hee; Lee, Je-Hwan; Lee, Kyoo-Hyung; Lee, Dae Ho; Suh, Cheolwon; Kim, Sung-Han

    2011-05-01

    There are limited data on the incidence of subsequent bloodstream infection (BSI) and the effect of systemic antibiotics in patients who had positive catheter-drawn blood cultures (CBC) and negative peripheral blood cultures (PBC). We retrospectively reviewed all paired blood cultures from patients with Hickman catheter in the hematology-oncology ward between January 1997 and December 2008. There were 112 episodes with positive CBC and negative PBC. Nine episodes (8.0%; 95% CI, 3.0-13.1%) led to subsequent BSI within 28 days. Subsequent BSI developed in 6 of 31 episodes (19%) where empiric antibiotics were inappropriate but in 3 of 81 episodes (4%) where empiric antibiotics were appropriate (P = 0.01). Subsequent candidemia (50%, 2 of 4) was more common than subsequent bacteremia (6%, 7 of 108) (P = 0.03). In conclusion, for patients with positive CBC and negative PBC, the overall incidence of subsequent BSI was 8.0%, and inappropriate empiric antibiotics was associated with subsequent BSI.

  18. Store-Operated Ca2+ Entry Does Not Control Proliferation in Primary Cultures of Human Metastatic Renal Cellular Carcinoma

    PubMed Central

    Turin, Ilaria; Potenza, Duilio Michele; Bottino, Cinzia; Glasnov, Toma N.; Ferulli, Federica; Mosca, Alessandra; Guerra, Germano; Rosti, Vittorio; Luinetti, Ombretta; Porta, Camillo; Pedrazzoli, Paolo

    2014-01-01

    Store-operated Ca2+ entry (SOCE) is activated following depletion of the inositol-1,4,5-trisphosphate (InsP3)-sensitive Ca2+ pool to regulate proliferation in immortalized cell lines established from either primary or metastatic lesions. The molecular nature of SOCE may involve both Stim1, which senses Ca2+ levels within the endoplasmic reticulum (ER) Ca2+ reservoir, and a number of a Ca2+-permeable channels on the plasma membrane, including Orai1, Orai3, and members of the canonical transient receptor (TRPC1–7) family of ion channels. The present study was undertaken to assess whether SOCE is expressed and controls proliferation in primary cultures isolated from secondary lesions of heavily pretreated metastatic renal cell carcinoma (mRCC) patients. SOCE was induced following pharmacological depletion of the ER Ca2+ store, but not by InsP3-dependent Ca2+ release. Metastatic RCC cells express Stim1-2, Orai1–3, and TRPC1–7 transcripts and proteins. In these cells, SOCE was insensitive to BTP-2, 10 µM Gd3+ and Pyr6, while it was inhibited by 100 µM Gd3+, 2-APB, and carboxyamidotriazole (CAI). Neither Gd3+ nor 2-APB or CAI impaired mRCC cell proliferation. Consistently, no detectable Ca2+ signal was elicited by growth factor stimulation. Therefore, a functional SOCE is expressed but does not control proliferation of mRCC cells isolated from patients resistant to multikinase inhibitors. PMID:25126575

  19. Phosphatidylserine exposure on stored red blood cells as a parameter for donor-dependent variation in product quality.

    PubMed

    Dinkla, Sip; Peppelman, Malou; Van Der Raadt, Jori; Atsma, Femke; Novotný, Vera M J; Van Kraaij, Marian G J; Joosten, Irma; Bosman, Giel J C G M

    2014-04-01

    Exposure of phosphatidylserine on the outside of red blood cells contributes to recognition and removal of old and damaged cells. The fraction of phosphatidylserine-exposing red blood cells varies between donors, and increases in red blood cell concentrates during storage. The susceptibility of red blood cells to stress-induced phosphatidylserine exposure increases with storage. Phosphatidylserine exposure may, therefore, constitute a link between donor variation and the quality of red blood cell concentrates. In order to examine the relationship between storage parameters and donor characteristics, the percentage of phosphatidylserine-exposing red blood cells was measured in red blood cell concentrates during storage and in fresh red blood cells from blood bank donors. The percentage of phosphatidylserine-exposing red blood cells was compared with red blood cell susceptibility to osmotic stress-induced phosphatidylserine exposure in vitro, with the regular red blood cell concentrate quality parameters, and with the donor characteristics age, body mass index, haemoglobin level, gender and blood group. Phosphatidylserine exposure varies between donors, both on red blood cells freshly isolated from the blood, and on red blood cells in red blood cell concentrates. Phosphatidylserine exposure increases with storage time, and is correlated with stress-induced phosphatidylserine exposure. Increased phosphatidylserine exposure during storage was found to be associated with haemolysis and vesicle concentration in red blood cell concentrates. The percentage of phosphatidylserine-exposing red blood cells showed a positive correlation with the plasma haemoglobin concentration of the donor. The fraction of phosphatidylserine-exposing red blood cells is a parameter of red blood cell integrity in red blood cell concentrates and may be an indicator of red blood cell survival after transfusion. Measurement of phosphatidylserine exposure may be useful in the selection of donors and

  20. Taking blood cultures from a newly established intravenous catheter in the emergency department does not increase the rate of contaminated blood cultures.

    PubMed

    Kelly, Anne-Maree; Klim, Sharon

    2013-10-01

    It has been suggested that blood cultures drawn from vascular catheters have a higher false positive rate than those drawn by venepuncture. In the face of institutionally imposed practice change prohibiting obtaining blood cultures from intravenous (i.v.) catheters in the ED, our aim of was to compare the rate of contaminated blood cultures between those taken from recently placed i.v. catheters and those taken by direct venepuncture. Prospective, non-randomised, observational study comparing the rate of contaminated blood cultures for specimens taken from recently placed (<1 h) i.v. catheters and direct venepuncture in adult ED patients. Outcome of interest was the rate of false positive cultures. Analysis was by comparison of proportions (χ(2) -test). Four hundred seventy-two blood culture sets were studied. There were 65 positive cultures, of which 49 (75%; 95% confidence interval [CI], 63-85%) were classified as true positive. The overall rate of contaminated blood cultures was 3.4% (95% CI, 2.0-5.6%). There was no difference in false positive rate between blood cultures taken via venepuncture and those taken from a recently placed i.v. cannula (P = 0.52; odds ratio, 0.9; 95% CI, 0.33-2.44). We found no difference in contaminated blood culture rate between recently placed i.v. catheters and direct venepuncture when infection control procedures were followed. © 2013 Australasian College for Emergency Medicine and Australasian Society for Emergency Medicine.

  1. Clinical Impact of Blood Culture Results in Acutely Ill Hospitalized Adult Patients With Cystic Fibrosis

    PubMed Central

    Vender, Robert J.; Vender, Robert L.

    2016-01-01

    Background Blood cultures are obtained clinically to confirm site and source of acute infection as well as to guide effective antibiotic therapies. Patients with cystic fibrosis (CF) are at risk for blood stream infection (BSI) as identified from positive blood culture results. Methods A retrospective chart review was performed of 190 adult CF patients from January 1, 2001 through December 1, 2015. All positive blood culture results were identified as to clinical relevance and source of BSI. Results There were a total of 3,053 blood cultures. One hundred fifty-one positive blood cultures were considered pathogenic and clinically significant. Venous access device-related BSI was identified in 31 evaluable patients and 106 blood cultures. Nineteen patients and 45 positive blood cultures were attributable to organ-specific sources. Conclusion Two patterns of BSI were identified: 1) venous access device infections without causal mortality and 2) organ-specific site infections with associated 26% mortality. PMID:27829951

  2. A change of culture: reducing blood culture contamination rates in an Emergency Department.

    PubMed

    Bentley, James; Thakore, Shobhan; Muir, L; Baird, Alastair; Lee, Jennifer

    2016-01-01

    Blood cultures are an important investigation to help tailor effective management for patients with severe sepsis. Frequent contaminated samples increase laboratory workload and can delay or cause incorrect changes to patient management. This can prolong patient hospitalisation, increase the risk of harm and increase cost to health boards. Current guidelines advocate a contamination rate of 2-3%. From January 2013 to November 2014 inclusive, the contamination rate was 4.74% in our Emergency Department, responsible for initial management and investigation of over 40 cases of sepsis per month. A Quality Improvement team was created to try to reduce contamination rates to the recommended target. An initial baseline survey of local staff showed good understanding of when to obtain a blood culture but there was variability in the methods and equipment used. A project was then conducted which focused on rationalising and standardising equipment and technique for blood culture sampling along with staff education to support this change. A simple department target of 30 days free from a contaminated blood culture was created which, if achieved, would ensure a contamination rate of less than 3%. This was supported by ongoing surveillance of contamination rates and investigation of contaminated sample cases. We were able to then identify high risk patients and factors which increased the chance of blood culture contamination. This allowed us to formulate solutions to help reduce the risks of contamination. Department achievements and learning points to help prevent further contamination were fed back positively to all staff. This project operated for 12-months and successfully reduced local contamination rates to 2.0%.

  3. In vitro parameters of cryopreserved leucodepleted and non-leucodepleted red blood cells collected by apheresis or from whole blood and stored in AS-3 for 21 days after thawing.

    PubMed

    Bohoněk, Miloš; Petráš, Marek; Turek, Ivo; Urbanová, Jaroslava; Hrádek, Tomáš; Staropražská, Věra; Koštířová, Jitka; Horčičková, Dana; Duchková, Simona

    2014-01-01

    The aim of the study was to evaluate the in vitro quality of cryopreserved red blood cells obtained from different sources with or without leucodepletion and stored at 4±2 °C in AS-3 for up to 21 days. Red blood cells were collected by four methods: double erythrocytapheresis, whole blood collection with buffy coat removal, double erythrocytapheresis with in-line leucofiltration, or whole blood collection with in-line leucofiltration. All four types of red blood cells were frozen in 40% glycerol after collection and stored at a temperature below -65 °C for at least 30 days, thawed, deglycerolised and subsequently reconstituted in AS-3. The in vitro haematological and biochemical properties of the thawed red blood cells were tested on days 0, 7, 14, and 21 after deglycerolisation and reconstitution. Overall, 72 units were processed. Leucodepletion of cryopreserved red blood cells units reduced haemolysis, lowered ammonia concentration, preserved pH and osmolality and led to sustained higher concentrations of ATP. In contrast, the source of red blood cells (apheresis or whole blood) did not affect their quality. The quality of all investigated red blood cells units was the same as or even better than that of erythrocytes obtained from double erythrocytapheresis with a 24-hour survival of at least 86% after up to 3 weeks of storage in AS-3.

  4. Monitoring infection: from blood culture to polymerase chain reaction (PCR).

    PubMed

    Book, Malte; Lehmann, Lutz Eric; Zhang, XiangHong; Stüber, Frank

    2013-06-01

    In patients with sepsis, diagnosis of blood stream infection (BSI) is a key concern to the therapist. Direct verification of pathogens in the blood stream executed by blood cultures (BC) still is regarded as the gold standard up to date. The quickest possible initiation of an appropriate antimicrobial therapy is a cornerstone of an effective therapy. Moreover, in this view BC can also serve to identify antimicrobial agents to target the pathogen. However, when employing BC the time needed until microbiological results are available ranges from 24 up to 72 h. Moreover, infections caused by multiple pathogens often remain undetected and concurrent antibiotic therapy may lower the overall sensitivity. Alternative pathogen characterization can be performed by polymerase chain reaction (PCR) based amplification methods. Results using PCR can be obtained within 6-8 h. Therefore, the time delay until an appropriate therapy can be reduced enormously. Moreover, these methods have the potential to enhance the sensitivity in the diagnosis of blood stream infections. Therefore, PCR based methods might be a valuable adjunct to present procedures of diagnosing bacteraemia.

  5. The cultural gradient: culture moderates the relationship between socioeconomic status (SES) and ambulatory blood pressure.

    PubMed

    Steffen, Patrick R

    2006-12-01

    A social gradient has been consistently demonstrated in Western countries with higher socioeconomic status (SES) related to lower blood pressure (BP). In non-Western countries, however, the social gradient is not always evident, with some countries appearing to show a reversed social gradient. It was hypothesized that culture moderates the social gradient, with the relationship between SES and BP differing as a function of culture. To investigate the idea of a "cultural gradient" a sample of Hispanic immigrants and Whites was studied. A total of 79 participants (30 Hispanic immigrant, 49 White) wore ambulatory blood pressure monitors for 24 h. The Hispanic immigrants also completed the Acculturation Rating Scale for Mexican Americans- II. Hispanic immigrants had lower SES and lower BP compared to Whites. A cultural gradient moderating the social gradient was evident with Hispanic immigrants displaying a positive relationship between SES and BP and Whites displaying a negative relationship. Among Hispanic immigrants, increased acculturation to Western culture decreased the positive relationship between SES and BP. Just as there is a social gradient with increasing socioeconomic status related to better cardiovascular health, there appears to be a cultural gradient with increasing acculturation to Western society related to worse cardiovascular health.

  6. Detection of Blood Culture Bacterial Contamination using Natural Language Processing

    PubMed Central

    Matheny, Michael E.; FitzHenry, Fern; Speroff, Theodore; Hathaway, Jacob; Murff, Harvey J.; Brown, Steven H.; Fielstein, Elliot M.; Dittus, Robert S.; Elkin, Peter L.

    2009-01-01

    Microbiology results are reported in semi-structured formats and have a high content of useful patient information. We developed and validated a hybrid regular expression and natural language processing solution for processing blood culture microbiology reports. Multi-center Veterans Affairs training and testing data sets were randomly extracted and manually reviewed to determine the culture and sensitivity as well as contamination results. The tool was iteratively developed for both outcomes using a training dataset, and then evaluated on the test dataset to determine antibiotic susceptibility data extraction and contamination detection performance. Our algorithm had a sensitivity of 84.8% and a positive predictive value of 96.0% for mapping the antibiotics and bacteria with appropriate sensitivity findings in the test data. The bacterial contamination detection algorithm had a sensitivity of 83.3% and a positive predictive value of 81.8%. PMID:20351890

  7. Comparison of ethanol and other drugs of abuse concentrations in whole blood stored in venoject glass and plastic and venosafe plastic evacuated tubes.

    PubMed

    Karinen, Ritva; Oiestad, Elisabeth Leere; Andresen, Wenche; Wethe, Grete; Smith-Kielland, Anne; Christophersen, Asbjørg

    2010-09-01

    The aim of this study was to evaluate the stability of blood concentrations of a variety of illegal and medicinal drugs that are important for forensic analyses when spiked and stored in Vacutainer or Venosafe evacuated plastic collection tubes compared to Vacutainer evacuated glass tubes. Tubes were filled with spiked whole blood and analyzed after storage for one week at ambient temperature and at -20 degrees C, respectively. Freeze-and-thaw stability was included in the study. No significant difference between storage in glass or plastic tubes was noted for any compound investigated.

  8. Plasma from stored packed red blood cells and MHC class I antibodies causes acute lung injury in a 2-event in vivo rat model

    PubMed Central

    Kelher, Marguerite R.; Masuno, Tomhiko; Moore, Ernest E.; Damle, Sagar; Meng, Xianzhong; Song, Yong; Liang, Xiayuan; Niedzinski, Jerry; Geier, Steven S.; Khan, Samina Y.; Gamboni-Robertson, Fabia

    2009-01-01

    Transfusion-related acute lung injury (TRALI) is the leading cause of transfusion death. We hypothesize that TRALI requires 2 events: (1) the clinical condition of the patient and (2) the infusion of antibodies against MHC class I antigens or the plasma from stored blood. A 2-event rat model was developed with saline (NS) or endotoxin (LPS) as the first event and the infusion of plasma from packed red blood cells (PRBCs) or antibodies (OX18 and OX27) against MHC class I antigens as the second event. ALI was determined by Evans blue dye leak from the plasma to the bronchoalveolar lavage fluid (BALF), protein and CINC-1 concentrations in the BALF, and the lung histology. NS-treated rats did not evidence ALI with any second events, and LPS did not cause ALI. LPS-treated animals demonstrated ALI in response to plasma from stored PRBCs, both prestorage leukoreduced and unmodified, and to OX18 and OX27, all in a concentration-dependent fashion. ALI was neutrophil (PMN) dependent, and OX18/OX27 localized to the PMN surface in vivo and primed the oxidase of rat PMNs. We conclude that TRALI is the result of 2 events with the second events consisting of the plasma from stored blood and antibodies that prime PMNs. PMID:19131548

  9. A study of the quantity of some stable and labile coagulation factors in fresh-frozen plasma produced from whole blood stored for 24 hours in Iran

    PubMed Central

    Naghadeh, Hossin Timori; Roudkenar, Mehryar Habibi

    2009-01-01

    Background The aim of this study was to assess whether the quantities of some coagulation factors in fresh-frozen plasma (FFP) produced from whole blood stored at 4°C for 24 h are adequate for their intended purpose. Materials and methods The amounts of some coagulation factors (fibrinogen, FV, FVII, FVIII, FX and FXI) in FFP separated from whole blood after storage at 4°C for 24 h were compared with the amounts of the corresponding coagulation factors in FFP separated from whole blood within 8 h of donation. Results In 98% of the FFP units prepared after 24 h of storage, the levels of fibrinogen, FV, FVII, FX and FXI were greater than 0.5 IU/mL. The concentration of FVIII in the 24 h plasma units was 82% of that found in the FFP units prepared within 8 h of blood collection. In FFP, FVIII, FVII and FX were reduced by 38%, 8% and 3%, respectively, but FV, FXI and fibrinogen were not reduced. Conclusion These data suggest that there is good retention of coagulation factor activity in plasma produced from whole blood stored at 4ºC for 24 h and that such plasma would be an acceptable product for most patients requiring FFP. PMID:19290079

  10. A study of the quantity of some stable and labile coagulation factors in fresh-frozen plasma produced from whole blood stored for 24 hours in Iran.

    PubMed

    Naghadeh, Hossin Timori; Roudkenar, Mehryar Habibi

    2009-01-01

    The aim of this study was to assess whether the quantities of some coagulation factors in fresh-frozen plasma (FFP) produced from whole blood stored at 4 degrees C for 24 h are adequate for their intended purpose. The amounts of some coagulation factors (fibrinogen, FV, FVII, FVIII, FX and FXI) in FFP separated from whole blood after storage at 4 degrees C for 24 h were compared with the amounts of the corresponding coagulation factors in FFP separated from whole blood within 8 h of donation. In 98% of the FFP units prepared after 24 h of storage, the levels of fibrinogen, FV, FVII, FX and FXI were greater than 0.5 IU/mL. The concentration of FVIII in the 24 h plasma units was 82% of that found in the FFP units prepared within 8 h of blood collection. In FFP, FVIII, FVII and FX were reduced by 38%, 8% and 3%, respectively, but FV, FXI and fibrinogen were not reduced. These data suggest that there is good retention of coagulation factor activity in plasma produced from whole blood stored at 4 masculineC for 24 h and that such plasma would be an acceptable product for most patients requiring FFP.

  11. Detection of African swine fever virus DNA in blood samples stored on FTA cards from asymptomatic pigs in Mbeya region, Tanzania.

    PubMed

    Braae, U C; Johansen, M V; Ngowi, H A; Rasmussen, T B; Nielsen, J; Uttenthal, Å

    2015-02-01

    The aim of the study was to assess whether blood samples collected onto FTA(®) cards could be used in combination with real-time PCR for the detection of African swine fever virus (ASFV) DNA in samples from resource-poor settings under the assumption that asymptomatically (sub-clinically) infected pigs may be present. Blood samples were collected from clinically healthy pigs from Mbeya Region, Tanzania. The blood samples were stored on FTA(®) cards and analysed by real-time PCR assays in duplicate; three pigs had high levels of viral DNA (Ct values of 27-29), and three pigs had a low level of viral DNA (Ct 36-45). Four pigs were positive in one of the duplicate samples only, but clear products of the expected size were obtained when the reactions were analysed by gel electrophoresis. For comparison, blood samples from pigs experimentally infected with either a pathogenic (OURT T88/1) or a non-pathogenic (OURT T88/3) isolate of ASFV were collected, stored on FTA(®) cards and analysed in the same way. The blood from pigs infected with the OURT T88/1 isolate showed high levels of viral DNA (Ct 22-33), whereas infection with non-pathogenic OURT T88/3 isolate resulted in only low levels of viral DNA (Ct 39) in samples collected at 10-14 days after inoculation.

  12. Effect of Storage Temperature on the Phenotype of Cultured Epidermal Cells Stored in Xenobiotic-Free Medium.

    PubMed

    Jackson, Catherine; Eidet, Jon R; Reppe, Sjur; Aass, Hans Christian D; Tønseth, Kim A; Roald, Borghild; Lyberg, Torstein; Utheim, Tor P

    2016-06-01

    Cultured epidermal cell sheets (CECS) are used in the treatment of large area burns to the body and have potential to treat limbal stem cell deficiency (LSCD) as shown in animal studies. Despite widespread use, storage options for CECS are limited. Short-term storage allows flexibility in scheduling surgery, quality control and improved transportation to clinics worldwide. Recent evidence points to the phenotype of cultured epithelial cells as a critical predictor of post-operative success following transplantation of CECS in burns and in transplantation of cultured epithelial cells in patients with LSCD. This study, therefore assessed the effect of a range of temperatures, spanning 4-37 °C, on the phenotype of CECS stored over a 2-week period in a xenobiotic-free system. Progenitor cell (p63, ΔNp63α and ABCG2) and differentiation (C/EBPδ and CK10) associated marker expression was assessed using immunocytochemistry. Immunohistochemistry staining of normal skin for the markers p63, ABCG2 and C/EBPδ was also carried out. Assessment of progenitor cell side population (SP) was performed using JC1 dye by flow cytometry. P63 expression remained relatively constant throughout the temperature range but was significantly lower compared to control between 20 and 28 °C (p < 0.05). High C/EBPδ together with low p63 suggested more differentiation beginning at 20 °C and above. Lower CK10 and C/EBPδ expression most similar to control was seen at 12 °C. The percentage of ABCG2 positive cells was most similar to control between 8 and 24 °C. Between 4 and 24 °C, the SP fluctuated, but was not significantly different compared to control. Results were supported by staining patterns indicating differentiation status associated with markers in normal skin sections. Lower storage temperatures, and in particular 12 °C, merit further investigation as optimal storage temperature for maintenance of undifferentiated phenotype in CECS.

  13. Molecular detection of antibiotic resistance genes from positive blood cultures.

    PubMed

    Hindiyeh, Musa Y; Smollan, Gill; Gefen-Halevi, Shiraz; Mendelson, Ella; Keller, Nathan

    2015-01-01

    Rapid detection of the bacterial causative agent causing sepsis must be coupled with rapid identification of the antibiotic resistant mechanism that the pathogen might possess. Real-time PCR (qPCR)-based assays have been extensively utilized in the clinical microbiology field as diagnostic tools for the rapid detection of specific nucleic acid (NA) targets. In this chapter, we will discuss the technical aspects of using an internally controlled qPCR assay for the rapid detection of Klebsiella pneumoniae carbapenemase gene (bla KPC) in positive Bactec blood culture bottles. The multiplex qPCR (bla KPC/RNase P) utilizes specific primers and probes for the detection of the bacterial carbapenem resistance mechanism, bla KPC gene, and the internal control RNase P. The internal control of the qPCR assay is vital for detecting any inhibitors that are well known to be present in the blood culture bottles. Rapid detection of the antibiotic resistant mechanism present in the bacterial pathogen causing sepsis can help in better managing patients' infection.

  14. Blood and marrow cultures as indicators of bone contamination in cadaver donors.

    PubMed

    Martinez, Octavio V; Buck, Billy E; Hernandez, Maribel; Malinin, Theodore

    2003-04-01

    Of 770 cadaver bone donors evaluated, 185 had positive blood or ilium marrow aspirate cultures. These donors were matched with an immediately preceding or subsequent donor with negative blood and marrow cultures. Donors with cultures positive for skin contaminants only were not included in the study. Samples of the blood and bone marrow, surface swab cultures, and cultures of tissue samples of the excised skeletal tissues were obtained at the time of tissue procurement. There were 88 (48%) donors with similar microbial species recovered from the blood and ilium marrow. These donors had a higher rate of positive bone cultures (30%) than donors with positive blood (15%) or marrow cultures only (11%) or donors with negative blood and marrow culture results (7.3%). Recovery of similar isolates from the blood and marrow had a positive predictive value of 72% for the isolation of the same types of organisms from the excised tissues compared with 38% for donors with positive blood cultures only. Although not absolute predictors of tissue contamination, combined blood and bone marrow cultures were more reliable indicators of tissue contamination than blood cultures alone.

  15. Pneumococcal pneumonia: differences according to blood culture results

    PubMed Central

    2014-01-01

    Background Bacteremia by Streptococcus pneumoniae has been traditionally associated with poor outcomes in patients with pneumonia; however, data on its impact on outcomes are limited and are sometimes contradictory. Methods We performed a prospective study in two hospitals in northern Spain in which cases diagnosed with pneumococcal pneumonia were selected from a cohort of hospitalized patients with pneumonia between January 2001 and July 2009. We compared patients with pneumococcal bacteremic pneumonia with those with pneumococcal non-bacteremic pneumonia. Results We compared 492 patients with negative blood culture and 399 with positive culture results. Host related factors were very similar in both groups. Severity of illness on admission measured by CURB-65 score was similar in both groups. Adjusted analysis showed a greater likelihood of septic shock during in-hospital course among patients with pneumococcal bacteremia (OR, 2.1; 95% CI, 1.2–3.5; P = 0.006). Likewise, patients with positive blood culture had greater in-hospital mortality (OR 2.1; 95% CI, 1.1 - -3.9; P = 0.02), 15-day mortality (OR 3.6; 95% CI, 1.7 - 7.4; P = 0.0006), and 30-day mortality (OR, 2.7; 95% CI, 1.5 - 5; P = 0.002). Conclusions Although host related factors and severity on admission were very similar in the two groups, bacteremic patients had worse in-hospital course and outcomes. Bacteraemia in pneumococcal pneumonia is of prognostic significance. PMID:25096919

  16. What proportion of Salmonella Typhi cases are detected by blood culture? A systematic literature review.

    PubMed

    Mogasale, Vittal; Ramani, Enusa; Mogasale, Vijayalaxmi V; Park, JuYeon

    2016-05-17

    Blood culture is often used in definitive diagnosis of typhoid fever while, bone marrow culture has a greater sensitivity and considered reference standard. The sensitivity of blood culture measured against bone marrow culture results in measurement bias because both tests are not fully sensitive. Here we propose a combination of the two cultures as a reference to define true positive S. Typhi cases. Based on a systematic literature review, we identified ten papers that had performed blood and bone marrow culture for S. Typhi in same subjects. We estimated the weighted mean of proportion of cases detected by culture measured against true S. Typhi positive cases using a random effects model. Of 529 true positive S. Typhi cases, 61 % (95 % CI 52-70 %) and 96 % (95 % CI 93-99 %) were detected by blood and bone marrow cultures respectively. Blood culture sensitivity was 66 % (95 % CI 56-75 %) when compared with bone marrow culture results. The use of blood culture sensitivity as a proxy measure to estimate the proportion of typhoid fever cases detected by blood culture is likely to be an underestimate. As blood culture sensitivity is used as a correction factor in estimating typhoid disease burden, epidemiologists and policy makers should account for the underestimation.

  17. Levels of factor VIII and factor IX in fresh-frozen plasma produced from whole blood stored at 4 °C overnight in Turkey

    PubMed Central

    Agus, Neval; Yilmaz, Nisel; Colak, Ayfer; Liv, Fatma

    2012-01-01

    Background The aim of this study was to assess whether the quantities of factor VIII and factor IX in fresh-frozen plasma produced from whole blood stored at 4 °C for 24 hours are adequate for their intended purpose. Materials and methods Fresh-frozen plasma separated from whole blood after storage at 4 °C overnight (24 hours from donation) was compared with plasma prepared 8 hours after donation using a standard method. The amounts of factor VIII and factor IX obtained with the two methods were compared. Results Compared to the levels of factor VIII and factor IX in plasma prepared within 8 hours of blood collection, the levels in plasma prepared after 24 hours of storage at 4 °C were 25% and 9% lower, respectively. Ninety percent of the factor VIII and 100% of the factor IX levels were above 0.5 IU/mL (standard haematology reference range) after 24 hours of storage. Discussion These data suggest that there is good retention of coagulation factor activity in plasma produced from whole blood stored at 4 ºC for 24 hours and that such plasma would be an acceptable product for most patients requiring fresh-frozen plasma. PMID:22153689

  18. Levels of factor VIII and factor IX in fresh-frozen plasma produced from whole blood stored at 4 °C overnight in Turkey.

    PubMed

    Agus, Neval; Yilmaz, Nisel; Colak, Ayfer; Liv, Fatma

    2012-04-01

    The aim of this study was to assess whether the quantities of factor VIII and factor IX in fresh-frozen plasma produced from whole blood stored at 4 °C for 24 hours are adequate for their intended purpose. Fresh-frozen plasma separated from whole blood after storage at 4 °C overnight (24 hours from donation) was compared with plasma prepared 8 hours after donation using a standard method. The amounts of factor VIII and factor IX obtained with the two methods were compared. Compared to the levels of factor VIII and factor IX in plasma prepared within 8 hours of blood collection, the levels in plasma prepared after 24 hours of storage at 4 °C were 25% and 9% lower, respectively. Ninety percent of the factor VIII and 100% of the factor IX levels were above 0.5 IU/mL (standard haematology reference range) after 24 hours of storage. These data suggest that there is good retention of coagulation factor activity in plasma produced from whole blood stored at 4 ºC for 24 hours and that such plasma would be an acceptable product for most patients requiring fresh-frozen plasma.

  19. Influence of Pre-Storage Irradiation on the Oxidative Stress Markers, Membrane Integrity, Size and Shape of the Cold Stored Red Blood Cells.

    PubMed

    Antosik, Adam; Czubak, Kamila; Gajek, Arkadiusz; Marczak, Agnieszka; Glowacki, Rafal; Borowczyk, Kamila; Zbikowska, Halina Malgorzata

    2015-05-01

    To investigate the extent of oxidative damage and changes in morphology of manually isolated red blood cells (RBCs) from whole blood, cold stored (up to 20 days) in polystyrene tubes and subjected to pre-storage irradiation (50 Gy) and to compare the properties of SAGM-preserved RBCs stored under experimental conditions (polystyrene tubes) with RBCs from standard blood bag storage. The percentage of hemolysis as well as the extracellular activity of LDH, thiobarbituric acid-reactive substances, reduced glutathione (GSH), and total antioxidant capacity (TAC) were measured. Changes in the topology of RBC membrane, shape, and size were evaluated by flow cytometry and judged against microscopy images. Irradiation caused significant LDH release as well as increased hemolysis and lipid peroxidation, GSH depletion, and reduction of TAC. Prolonged storage of irradiated RBCs resulted in phosphatidylserine exposure on the cell surface. By day 20, approximately 60% of RBCs displayed non-discoid shape. We did not notice significant differences in percentage of altered cells and cell volume between RBCs exposed to irradiation and those not exposed. Irradiation of RBC transfusion units with a dose of 50 Gy should be avoided. For research purposes such as studying the role of antioxidants, storage of small volumes of RBCs derived from the same donor would be more useful, cheaper, and blood-saving.

  20. Bacillus cereus from blood cultures: virulence genes, antimicrobial susceptibility and risk factors for blood stream infection.

    PubMed

    Horii, Toshinobu; Notake, Shigeyuki; Tamai, Kiyoko; Yanagisawa, Hideji

    2011-11-01

    We characterized the profiles of virulence genes and antimicrobial susceptibility of Bacillus cereus isolates from blood cultures as well as the risk factors for blood stream infections (BSIs). The diversity of virulence gene patterns was found to be wide among 15 B. cereus isolates from BSIs and also among 11 isolates from contaminated blood cultures. The MicroScan broth microdilution method yielded results corresponding with those of the agar dilution (reference) method for levofloxacin, linezolid, and vancomycin, while the Etest results were consistent with the reference results for clindamycin, gentamicin, imipenem, levofloxacin, and linezolid. Compared with the reference values, however, some isolates showed marked differences of the minimum inhibitory concentrations (MICs) for ampicillin and clindamycin when determined using the MicroScan method, or the MICs for ampicillin, meropenem, and vancomycin when determined using the Etest method. Significantly more patients were treated with antimicrobials for more than 3 days during the 3-month period before isolation in the BSI group. Prior antimicrobial therapy may be a risk factor for BSIs due to B. cereus.

  1. Blood Culture Bottle and Standard Culture Bottle Methods for Detection of Bacterial Pathogens in Parapneumonic Pleural Effusion

    PubMed Central

    Charoentunyarak, Surapan; Kananuraks, Sarassawan; Chindaprasirt, Jarin; Limpawattana, Panita; Sawanyawisuth, Kittisak

    2015-01-01

    Background: Bacterial parapneumonic pleural effusions (PPEs) have high morbidity. The accurate identification of pathogens is vital for initiating the appropriate treatment. A previous study suggested that the use of blood culture bottles might improve the bacterial yield in PPEs. Objectives: The aim of this study was to compare the culture positivity rate by the blood culture bottles and the standard culture bottles in bacterial PPEs. Patients and Methods: Patients diagnosed with PPEs at the Khon Kaen Hospital, Khon Kaen, Thailand, which is an endemic area of melioidosis, were enrolled consecutively and prospectively. The study period was from June first, 2012 to December 31st, 2013. The inclusion criteria were adult patients aged > 18 years, with exudative, neutrophilic parapneumonic effusion. Of the pleural fluid samples, 5 mL from all the eligible patients were collected in both blood culture bottles and the standard culture bottles. Patient baseline characteristics, laboratory results, and culture results were collected and analyzed. Results: During the study period, 129 patients met the study criteria. The bacteria-positive rate of pleural fluid culture using the standard culture bottle was 14.0%, whereas the positive rate using blood culture bottles was 24.0% (P < 0.001). Conclusions: The blood culture bottle method is more effective than the standard culture bottle method for the detection of bacterial pathogens in PPE. PMID:26587217

  2. Blood culture status in mature horses with diarrhoea: a possible association with survival.

    PubMed

    Johns, I; Tennent-Brown, B; Schaer, B Dallap; Southwood, L; Boston, R; Wilkins, P

    2009-02-01

    The incidence and implications of positive blood cultures in mature horses with diarrhoea is unknown. The diagnosis of bacteraemia may alter treatment and prognosis. The proportion of horses with diarrhoea that are blood culture positive is higher than previously assumed and a positive blood culture has a negative impact on survival. Blood cultures were taken at admission and 24 h after admission from 31 mature horses with diarrhoea. Nine (29%) horses were blood culture positive within 24 h of admission. Organisms isolated included Corynebacterium spp. (n = 6), Streptococcus spp. (n = 2), Pantoea agglomerans (n = 1), Gram-negative rod (n = 1), Bacillus spp. (n = 1) and yeast (n = 1). Horses with positive blood cultures were significantly less likely to survive. Prior treatment with antimicrobial drugs had no significant effect on blood culture status. Horses with positive blood cultures had a significantly higher heart rate, packed cell volume (PCV) and plasma potassium concentration at admission, and a higher PCV and lower total plasma protein concentration 24 h after admission. Positive blood cultures occur more frequently than previously reported, and may have a negative impact on survival in horses with diarrhoea. Results of blood cultures may be useful in formulating a prognosis for horses with diarrhoea. Further research is required to determine the effect of antimicrobial treatment on outcome.

  3. Storage of platelet concentrates from pooled buffy coats made of fresh and overnight-stored whole blood processed on the novel Atreus 2C+ system: in vitro study.

    PubMed

    Sandgren, Per; Callaert, Martine; Shanwell, Agneta; Gulliksson, Hans

    2008-04-01

    The Atreus 2C+ system (Gambro BCT) automatically separates whole blood (WB) into buffy coat (BC), red blood cells (RBC), and plasma and transfers the components into separate containers. After processing with the Atreus, 4 to 6 BC units can be pooled and processed into leukoreduced platelets (PLTs) by use of the automated OrbiSac BC system (Gambro BCT). The aim of our in vitro study was to investigate the effects of holding either WB or BC overnight before preparation of PLTs by use of the Atreus 2C+ system for BC preparation. A standard routine procedure involving conventional blood containers for the preparation of BC combined with the OrbiSac process (top-and-top system; Terumo) was used as a reference. WB was either processed within 8 hours after collection ("fresh blood") or stored overnight before processing. WB units were separated into BC, RBC, and plasma units and transferred into individual containers. Either the BC or the WB units rested overnight at 22 +/- 2 degrees C. Six ABO-identical BCs, obtained from either fresh or overnight-stored WB, were pooled and processed with the OrbiSac BC system to obtain leukoreduced PLTs. In total, 20 Atreus and 10 reference (leukoreduced PLTs) samples were analyzed for various in vitro variables during the 7-day storage period. No significant difference in glucose consumption, lactate production, mean PLT volume, LDH activity, bicarbonate, ATP, RANTES, and the expression of CD62p and CD42b between groups was detected. pH was maintained at greater than 7.0 (Day 7). Swirling remained at the highest levels (score, 2) for all units throughout storage. PLTs derived from BCs, obtained from either fresh or overnight-stored WB processed on the novel automated Atreus 2C+ system, were equivalent to control PLTs with regard to PLT in vitro characteristics during 7 days of storage. Stable recovery of PLTs and satisfactory PLT content according to current standards were also found.

  4. Data on how several physiological parameters of stored red blood cells are similar in glucose 6-phosphate dehydrogenase deficient and sufficient donors.

    PubMed

    Tzounakas, Vassilis L; Kriebardis, Anastasios G; Georgatzakou, Hara T; Foudoulaki-Paparizos, Leontini E; Dzieciatkowska, Monika; Wither, Matthew J; Nemkov, Travis; Hansen, Kirk C; Papassideri, Issidora S; D'Alessandro, Angelo; Antonelou, Marianna H

    2016-09-01

    This article contains data on the variation in several physiological parameters of red blood cells (RBCs) donated by eligible glucose-6-phosphate dehydrogenase (G6PD) deficient donors during storage in standard blood bank conditions compared to control, G6PD sufficient (G6PD(+)) cells. Intracellular reactive oxygen species (ROS) generation, cell fragility and membrane exovesiculation were measured in RBCs throughout the storage period, with or without stimulation by oxidants, supplementation of N-acetylcysteine and energy depletion, following incubation of stored cells for 24 h at 37 °C. Apart from cell characteristics, the total or uric acid-dependent antioxidant capacity of the supernatant in addition to extracellular potassium concentration was determined in RBC units. Finally, procoagulant activity and protein carbonylation levels were measured in the microparticles population. Further information can be found in "Glucose 6-phosphate dehydrogenase deficient subjects may be better "storers" than donors of red blood cells" [1].

  5. Coagulation function of stored whole blood is preserved for 14 days in austere conditions: A ROTEM feasibility study during a Norwegian antipiracy mission and comparison to equal ratio reconstituted blood.

    PubMed

    Strandenes, Geir; Austlid, Ivar; Apelseth, Torunn O; Hervig, Tor A; Sommerfelt-Pettersen, Jan; Herzig, Maryanne C; Cap, Andrew P; Pidcoke, Heather F; Kristoffersen, Einar K

    2015-06-01

    Formulation of a medical preparedness plan for treating severely bleeding casualties during naval deployment is a significant challenge because of territory covered during most missions. The aim of this study was to evaluate the concept of "walking blood bank" as a supportable plan for supplying safe blood and blood products. In 2013, the Royal Norwegian Navy conducted antipiracy operations from a frigate, beginning in the Gulf of Aden and ending in the Indian Ocean. Crews were on 24-hour emergency alert in preparation for an enemy assault on the frigate. Under an approved command protocol, a "walking blood bank," using crew blood donations, was established for use on board and on missions conducted in rigid-hulled inflatable boats, during which freeze-dried plasma and leukoreduced, group O low anti-A/anti-B titer, cold-stored whole blood were stored in Golden Hour Boxes. Data demonstrating the ability to collect, store, and provide whole blood were collected to establish feasibility of implementing a whole blood-focused remote damage-control resuscitation program aboard a naval vessel. In addition, ROTEM data were collected to demonstrate feasibility of performing this analysis on a large naval vessel and to also measure hemostatic efficacy of cold-stored leukoreduced whole blood (CWB) stored during a period of 14 days. ROTEM data on CWB was compared with reconstituted whole blood. Drills simulating massive transfusion activation were conducted, in which 2 U of warm fresh whole blood with platelet sparing leukoreduction were produced in 40 minutes, followed by collection of two additional units at 15-minute increments. The ROTEM machine performed well during ship-rolling, as shown by the overlapping calculated and measured mechanical piston movements measured by the ROTEM device. Error messages were recorded in 4 (1.5%) of 267 tests. CWB yielded reproducible ROTEM results demonstrating preserved fibrinogen function and platelet function for at least 3.5 weeks and

  6. Can We Reduce Negative Blood Cultures With Clinical Scores and Blood Markers? Results From an Observational Cohort Study.

    PubMed

    Laukemann, Svenja; Kasper, Nina; Kulkarni, Prasad; Steiner, Deborah; Rast, Anna Christina; Kutz, Alexander; Felder, Susan; Haubitz, Sebastian; Faessler, Lukas; Huber, Andreas; Fux, Christoph A; Mueller, Beat; Schuetz, Philipp

    2015-12-01

    Only a small proportion of blood cultures routinely performed in emergency department (ED) patients is positive. Multiple clinical scores and biomarkers have previously been examined for their ability to predict bacteremia. Conclusive clinical validation of these scores and biomarkers is essential.This observational cohort study included patients with suspected infection who had blood culture sampling at ED admission. We assessed 5 clinical scores and admission concentrations of procalcitonin (PCT), C-reactive protein (CRP), lymphocyte and white blood cell counts, the neutrophil-lymphocyte count ratio (NLCR), and the red blood cell distribution width (RDW). Two independent physicians assessed true blood culture positivity. We used logistic regression models with area under the curve (AUC) analysis.Of 1083 patients, 104 (9.6%) had positive blood cultures. Of the clinical scores, the Shapiro score performed best (AUC 0.729). The best biomarkers were PCT (AUC 0.803) and NLCR (AUC 0.700). Combining the Shapiro score with PCT levels significantly increased the AUC to 0.827. Limiting blood cultures only to patients with either a Shapiro score of ≥4 or PCT > 0.1 μg/L would reduce negative sampling by 20.2% while still identifying 100% of positive cultures. Similarly, a Shapiro score ≥3 or PCT >0.25 μg/L would reduce cultures by 41.7% and still identify 96.1% of positive blood cultures.Combination of the Shapiro score with admission levels of PCT can help reduce unnecessary blood cultures with minimal false negative rates.The study was registered on January 9, 2013 at the 'ClinicalTrials.gov' registration web site (NCT01768494).

  7. Can We Reduce Negative Blood Cultures With Clinical Scores and Blood Markers? Results From an Observational Cohort Study

    PubMed Central

    Laukemann, Svenja; Kasper, Nina; Kulkarni, Prasad; Steiner, Deborah; Rast, Anna Christina; Kutz, Alexander; Felder, Susan; Haubitz, Sebastian; Faessler, Lukas; Huber, Andreas; Fux, Christoph A.; Mueller, Beat; Schuetz, Philipp

    2015-01-01

    Abstract Only a small proportion of blood cultures routinely performed in emergency department (ED) patients is positive. Multiple clinical scores and biomarkers have previously been examined for their ability to predict bacteremia. Conclusive clinical validation of these scores and biomarkers is essential. This observational cohort study included patients with suspected infection who had blood culture sampling at ED admission. We assessed 5 clinical scores and admission concentrations of procalcitonin (PCT), C-reactive protein (CRP), lymphocyte and white blood cell counts, the neutrophil-lymphocyte count ratio (NLCR), and the red blood cell distribution width (RDW). Two independent physicians assessed true blood culture positivity. We used logistic regression models with area under the curve (AUC) analysis. Of 1083 patients, 104 (9.6%) had positive blood cultures. Of the clinical scores, the Shapiro score performed best (AUC 0.729). The best biomarkers were PCT (AUC 0.803) and NLCR (AUC 0.700). Combining the Shapiro score with PCT levels significantly increased the AUC to 0.827. Limiting blood cultures only to patients with either a Shapiro score of ≥4 or PCT > 0.1 μg/L would reduce negative sampling by 20.2% while still identifying 100% of positive cultures. Similarly, a Shapiro score ≥3 or PCT >0.25 μg/L would reduce cultures by 41.7% and still identify 96.1% of positive blood cultures. Combination of the Shapiro score with admission levels of PCT can help reduce unnecessary blood cultures with minimal false negative rates. The study was registered on January 9, 2013 at the ‘ClinicalTrials.gov’ registration web site (NCT01768494). PMID:26656373

  8. Presence of blood-sucking mesostigmatic mites in rodents and birds kept in pet stores in the Cracow area, Poland.

    PubMed

    Kowal, Jerzy; Nosal, Paweł; Niedziółka, Rafał; Kornaś, Sławomir

    2014-01-01

    The aim of the study was to investigate the occurrence of zoonotic arthropod parasites in small animals sold in selected pet stores in the Cracow area. The research was conducted in seven pet stores, keeping a total of six species of rodents and three species of birds. In two shops, two species of mites of the order Mesostigmata were detected on the animals and in their surrounding: Dermanyssus gallinae, the poultry red mite, and Ornithonyssus bacoti, the rat mite. Both observed species of mites may be harmful to animals, as well as to people working in the shops or potential pet owners. This study discusses the possible origin of the parasites and their importance to human health.

  9. Simultaneous Two-site Blood Culture for Diagnosis of Neonatal Sepsis.

    PubMed

    Tomar, Priya; Garg, Amit; Gupta, Rashee; Singh, Abhishek; Gupta, Navratan Kumar; Upadhyay, Amit

    2017-03-15

    To evaluate efficacy of two blood cultures taken simultaneously from two different sites as compared to standard practice of single blood culture in diagnosis of neonatal sepsis. Prospective cohort study. A tertiary-care center at a public hospital. 475 neonates admitted to intensive care unit with suspected sepsis, from August 2014-July 2015. Two blood cultures drawn from two different peripheral veins in patients with suspected neonatal sepsis. Increase in culture-positivity rate with use of two blood cultures. 475 babies with suspected sepsis were enrolled. 185 patients had only first culture positive (38.9%). When we added second culture positivity, yield increased to 221 (46.5%). Adding on second culture increased the culture yield by 36 (7.6%; 95% CI 2.41 to 12.79; P=0.018). The most common organisms isolated were E. coli, S. aureus and Candida spp. Major morbidities and mortality were more common in blood culture positive patients Contamination was ruled out in 25 babies who grew Coagulase negative Staphylococcus (CONS) (n=10) and Candida spp. (n=15) in either of the two cultures. Two blood cultures taken simultaneously from two different sites improve rate of pathogen detection as compared to routine practice of single blood culture.

  10. The design and evaluation of a shaped filter collection device to sample and store defined volume dried blood spots from finger pricks.

    PubMed

    Polley, Spencer D; Bell, David; Oliver, James; Tully, Frank; Perkins, Mark D; Chiodini, Peter L; González, Iveth J

    2015-02-05

    Dried blood spots are a common medium for collecting patient blood prior to testing for malaria by molecular methods. A new shaped filter device for the quick and simple collection of a designated volume of patient blood has been designed and tested against conventional blood spots for accuracy and precision. Shaped filter devices were laser cut from Whatman GB003 paper to absorb a 20 μl blood volume. These devices were used to sample Plasmodium falciparum infected blood and the volume absorbed was measured volumetrically. Conventional blood spots were made by pipetting 20 μl of the same blood onto Whatman 3MM paper. DNA was extracted from both types of dried blood spot using Qiagen DNA blood mini or Chelex extraction for real-time PCR analysis, and PURE extraction for malaria LAMP testing. The shaped filter devices collected a mean volume of 21.1 μl of blood, with a coefficient of variance of 8.1%. When used for DNA extraction by Chelex and Qiagen methodologies the mean number of international standard units of P. falciparum DNA recovered per μl of the eluate was 53.1 (95% CI: 49.4 to 56.7) and 32.7 (95% CI: 28.8 to 36.6), respectively for the shaped filter device, and 54.6 (95% CI: 52.1 to 57.1) and 12.0 (95% CI: 9.9 to 14.1), respectively for the 3MM blood spots. Qiagen extraction of 200 μl of whole infected blood yielded 853.6 international standard units of P. falciparum DNA per μl of eluate. A shaped filter device provides a simple way to quickly sample and store a defined volume of blood without the need for any additional measuring devices. Resultant dried blood spots may be employed for DNA extraction using a variety of technologies for nucleic acid amplification without the need for repeated cleaning of scissors or punches to prevent cross contamination of samples and results are comparable to traditional DBS.

  11. Blood Cultures at Central Line Insertion in the Intensive Care Unit: Comparison with Peripheral Venipuncture▿

    PubMed Central

    Stohl, Sheldon; Benenson, Shmuel; Sviri, Sigal; Avidan, Alexander; Block, Colin; Sprung, Charles L.; Levin, Phillip D.

    2011-01-01

    Blood cultures are a key diagnostic test for intensive care unit (ICU) patients; however, contaminants complicate interpretations and lead to unnecessary antibiotic administration and costs. Indications for blood cultures and central venous catheter (CVC) insertions often overlap for ICU patients. Obtaining blood cultures under the strict sterile precautions utilized for CVC insertion might be expected to decrease culture contamination. This retrospective study compared the results of blood cultures taken at CVC insertion, at arterial line insertion, and from peripheral venipuncture in order to validate the advantage of CVC insertion cultures. Cultures from indwelling lines were excluded. Results of 14,589 blood cultures, including 2,736 (19%) CVC, 1,513 (10%) arterial line, and 10,340 (71%) peripheral cultures taken over 5.5 years in two ICUs (general and medical) were analyzed. CVC cultures were contaminated more frequently than arterial line or peripheral cultures (225/2,736 [8%] CVC, 48/1,513 [3%] arterial line, and 378/10,340 (4%) peripheral cultures [P < 0.001 for CVC versus peripheral and CVC versus arterial line cultures]). True pathogens were found more frequently in CVC insertion cultures (334/2,736 [12%] CVC, 155/1,513 [10%] arterial line, and 795/10,340 [8%] peripheral cultures [P < 0.001 for CVC versus peripheral cultures; P = 0.055 for CVC versus arterial line cultures; P < 0.001 for peripheral versus arterial line cultures]). Contamination and true-positive rates were similar for culture sets from the two ICUs for each given culture source. Despite superior sterile precautions, cultures taken at the time of central line insertion had a higher contamination rate than did either peripheral or arterial line blood cultures. This may be related to the increased manipulations required for CVC insertion. PMID:21525219

  12. Evaluation of the Sterifil lysis-filtration blood culture system.

    PubMed

    Farmer, S G; Komorowski, R A

    1972-03-01

    This paper describes the comparison of the Sterifil lysis-filtration (SLF) blood culture procedure with a standard Trypticase soy broth (TSB) technique. The lysing solutions employed in the SLF system, Triton X-100 (alkyl phenoxy polyethoxy ethanol) and sodium carbonate, were deleterious to most bacteria commonly encountered in bacteremia except staphylococci and enterococci. Candida was not adversely affected. There was a positive correlation between the tolerance of the microbial isolants to the lysing solutions and their recovery by the SLF technique. A total of 3,554 cultures were run in parallel and 398 isolants were obtained. Of 201 gram-positive isolants, 135 were recovered by both techniques, 43 were detected by the TSB technique only, and 23 were recovered only with the SLF method. In sharp contrast, of 168 gram-negative isolants, 28 were recovered in common, 130 were isolated only by TSB, and 10 were recovered only with the SLF method. The SLF method detected all cases of candidemia detected by the TSB method plus an additional 12 for a total of 29 cases. The SLF method, as currently described, is generally too toxic to bacteria for routine use in a clinical laboratory.

  13. Impact of Pre-Analytical Time on the Recovery of Pathogens from Blood Cultures: Results from a Large Retrospective Survey

    PubMed Central

    Borsari, Lucia; Aggazzotti, Gabriella; Busani, Stefano; Mussini, Cristina; Rumpianesi, Fabio; Rossolini, Gian Maria; Girardis, Massimo

    2017-01-01

    Background Prompt identification of bloodstream pathogens is essential for optimal management of patients. Significant changes in analytical methods have improved the turnaround time for laboratory diagnosis. Less attention has been paid to the time elapsing from blood collection to incubation and to its potential effect on recovery of pathogens. We evaluated the performance of blood cultures collected under typical hospital conditions in relation to the length of their pre-analytical time. Methods We carried out a large retrospective study including 50,955 blood cultures collected, over a 30-month period, from 7,035 adult septic patients. Cultures were accepted by the laboratory only during opening time (Mon-Fri: 8am–4pm; Sat: 8am–2pm). Samples collected outside laboratory hours were stored at room temperature at clinical wards. All cultures were processed by automated culture systems. Day and time of blood collection and of culture incubation were known for all samples. Results A maximum pre-analytical interval of 2 hours is recommended by guidelines. When the laboratory was open, 57% of cultures were processed within 2 h. When the laboratory was closed, 4.9% of cultures were processed within 2 h (P<0.001). Samples collected when the laboratory was closed showed pre-analytical times significantly longer than those collected when laboratory was open (median time: 13 h and 1 h, respectively, P<0.001). The prevalence of positive cultures was significantly lower for samples collected when the laboratory was closed compared to open (11% vs 13%, P<0.001). The probability of a positive result decreased of 16% when the laboratory was closed (OR:0.84; 95%CI:0.80–0.89, P<0.001). Further, each hour elapsed from blood collection to incubation resulted associated with a decrease of 0.3% (OR:0.997; 95%CI:0.994–0.999, P<0.001) in the probability of a positive result. Discussion Delayed insertions of cultures into automated systems was associated with lower detection

  14. Reintoxication: the release of fat-stored Δ9-tetrahydrocannabinol (THC) into blood is enhanced by food deprivation or ACTH exposure

    PubMed Central

    Gunasekaran, N; Long, LE; Dawson, BL; Hansen, GH; Richardson, DP; Li, KM; Arnold, JC; McGregor, IS

    2009-01-01

    Background and purpose: Δ9-tetrahydrocannabinol (THC), the main psychoactive constituent of cannabis, accumulates in adipose tissue where it is stored for long periods of time. Here we investigated whether conditions that promote lipolysis can liberate THC from adipocytes to yield increased blood levels of THC. Experimental approach: In vitro studies involved freshly isolated rat adipocytes that were incubated with THC before exposure to the lipolytic agent adrenocorticotrophic hormone (ACTH). A complementary in vivo approach examined the effects of both food deprivation and ACTH on blood levels of THC in rats that had been repeatedly injected with THC (10 mg·kg−1) for 10 consecutive days. Lipolysis promoted by ACTH or food deprivation was indexed by measurement of glycerol levels. Key results: ACTH increased THC levels in the medium of THC-pretreated adipocytes in vitro. ACTH also enhanced THC release from adipocytes in vitro when taken from rats repeatedly pretreated with THC in vivo. Finally, in vivo ACTH exposure and 24 h food deprivation both enhanced the levels of THC and its metabolite, (-)-11-nor-9-carboxy-Δ9-tetrahydrocannabinol (THC-COOH) in the blood of rats that had been pre-exposed to repeated THC injections. Conclusions and implications: The present study shows that lipolysis enhances the release of THC from fat stores back into blood. This suggests the likelihood of ‘reintoxication’ whereby food deprivation or stress may raise blood THC levels in animals chronically exposed to the drug. Further research will need to confirm whether this can lead to functional effects, such as impaired cognitive function or ‘flashbacks’. PMID:19681888

  15. Utility of surveillance blood cultures in patients undergoing hematopoietic stem cell transplantation

    PubMed Central

    2014-01-01

    Background Surveillance blood cultures are often obtained in hematopoietic stem cell transplant (HSCT) patients for detection of bloodstream infection. The major aims of this retrospective cohort study were to determine the utility of the practice of obtaining surveillance blood cultures from asymptomatic patients during the first 100 post-transplant days and to determine if obtaining more than one positive blood culture helps in the diagnosis of bloodstream infection. Methods We conducted a 17-month retrospective analysis of all blood cultures obtained for patients admitted to the hospital for HSCT from January 2010 to June 2011. Each patient’s clinical course, vital signs, diagnostic testing, treatment, and response to treatment were reviewed. The association between number of positive blood cultures and the final diagnosis was analyzed. Results Blood culture results for 205 patients were reviewed. Cultures obtained when symptoms of infection were present (clinical cultures) accounted for 1,033 culture sets, whereas 2,474 culture sets were classified as surveillance cultures (no symptoms of infection were present). The total number of positive blood cultures was 185 sets (5.3% of cultures obtained) and accounted for 84 positive culture episodes. Incidence of infection in autologous, related allogeneic and unrelated allogeneic transplants was 8.3%, 20.0%, and 28.6% respectively. Coagulase-negative staphylococci were the most common organisms isolated. Based on our application of predefined criteria there were 29 infections and 55 episodes of positive blood cultures that were not infections. None of the patients who developed infection were diagnosed by surveillance blood cultures. None of the uninfected patients with positive blood cultures showed any clinical changes after receiving antibiotics. There was a significant difference between the incidence of BSI in the first and second 50-day periods post-HSCT. There was no association between the number of

  16. Enhancement of recovery of Neisseria meningitidis by gelatin in blood culture media.

    PubMed Central

    Pai, C H; Sorger, S

    1981-01-01

    The efficacy of gelatin for the recovery of Neisseria meningitidis from blood cultures was evaluated in a clinical setting. The organism was isolated from seven patients with meningococcal infections in blood culture media containing 1% gelatin. In contrast, only two blood cultures from these patients were positive in media without gelatin (P less than 0.05). Gelatin did not influence the recovery of other organisms isolated during this study. Conventional blood culture media may be supplemented with gelatin when meningococcemia is suspected. PMID:6790567

  17. Diagnostic yield of blood clot culture in the accurate diagnosis of enteric fever and human brucellosis.

    PubMed

    Mantur, Basappa G; Bidari, Laxman H; Akki, Aravind S; Mulimani, Mallanna S; Tikare, Nitin V

    2007-01-01

    Culture of blood is the most frequent, accurate means of diagnosing bacteremia in enteric fever and brucellosis. However, conventional blood culturing is slow in isolating bacteria causing these diseases. In this work, we evaluated the performance of blood clot culture and conventional whole blood cultures in the accurate diagnosis of enteric fever (253 cases) and human brucellosis (71cases). The blood clot culture was found to be much more sensitive for both Salmonella (more by 34.4%, P< 0.001) and Brucella (more by 22.6%, P<0.001) than whole blood culture. Bacterial growth was significantly faster in cultures of blood clot compared to whole blood (1.1 versus 2.6 days for Salmonella, 3.1 versus 8.2 days for Brucella melitensis, respectively). The rapid confirmation of the etiological agent would facilitate an early institution of appropriate antimicrobial therapy, thereby reducing clinical morbidity especially in an endemic population. It is worthwile practicing blood clot culture for the accurate diagnosis of enteric fever and brucellosis in developing countries where diagnostic facilities by advanced technologies like automated culture systems and PCR are not available.

  18. Predicting positive blood cultures in patients presenting with pneumonia at an Emergency Department in Singapore.

    PubMed

    Cham, Gregory; Yan, Sun; Heng, Bee Hoon; Seow, Eillyne

    2009-06-01

    Routine blood cultures have been recommended for all patients in treatment guidelines for community-acquired pneumonia (CAP). This practice has become a major area of resource utilisation, despite the lack of evidence in its clinical utility. Calls for abandoning the practice is balanced by the occasions of uncovering an unexpected pathogen or an unusual antimicrobial resistance pattern. The aim of this study is to identify factors that predict positive blood cultures among patients hospitalised for pneumonia upon presentation at the Emergency Department (ED). A case control study was carried out on patients treated for pneumonia in the ED who had routine blood cultures performed as part of their management. The pneumonia severity index (PSI) was used to categorize patients into low- and high-risk for 30-day mortality. Logistic regression was carried out to determine factors significantly associated with positive blood cultures, from which a predictive probability equation was used to identify patients whose blood cultures were negative at a pre-determined cut-off, with minimum number of culture positive misclassification. A scoring system was devised, with scores predicting which patients would be likely to have a positive or negative blood culture. A total of 1407 patients with pneumonia were treated at ED from May to December 2006, from whom 1800 blood cultures were performed. Of these, 140 cultures (7.8%) grew organisms, comprising 96 (5.3%) true positive cultures and 44 (2.4%) contaminated cultures. Logistic regression analysis identified ill patients with higher PSI classes, smokers and Malay patients to be more likely to have positive blood cultures. Patients who had prior treatment with antibiotics, chronic obstructive pulmonary disease and cough were less likely to have positive blood cultures. An index to predict a negative blood culture resulted in the accurate classification of all but 4 positive patients while still correctly classifying 27.8% of blood

  19. Transfusion of human volunteers with older, stored red blood cells produces extravascular hemolysis and circulating non-transferrin-bound iron.

    PubMed

    Hod, Eldad A; Brittenham, Gary M; Billote, Genia B; Francis, Richard O; Ginzburg, Yelena Z; Hendrickson, Jeanne E; Jhang, Jeffrey; Schwartz, Joseph; Sharma, Shruti; Sheth, Sujit; Sireci, Anthony N; Stephens, Hannah L; Stotler, Brie A; Wojczyk, Boguslaw S; Zimring, James C; Spitalnik, Steven L

    2011-12-15

    Transfusions of RBCs stored for longer durations are associated with adverse effects in hospitalized patients. We prospectively studied 14 healthy human volunteers who donated standard leuko-reduced, double RBC units. One unit was autologously transfused "fresh" (3-7 days of storage), and the other "older" unit was transfused after 40 to 42 days of storage. Of the routine laboratory parameters measured at defined times surrounding transfusion, significant differences between fresh and older transfusions were only observed in iron parameters and markers of extravascular hemolysis. Compared with fresh RBCs, mean serum total bilirubin increased by 0.55 mg/dL at 4 hours after transfusion of older RBCs (P = .0003), without significant changes in haptoglobin or lactate dehydrogenase. In addition, only after the older transfusion, transferrin saturation increased progressively over 4 hours to a mean of 64%, and non-transferrin-bound iron appeared, reaching a mean of 3.2μM. The increased concentrations of non-transferrin-bound iron correlated with enhanced proliferation in vitro of a pathogenic strain of Escherichia coli (r = 0.94, P = .002). Therefore, circulating non-transferrin-bound iron derived from rapid clearance of transfused, older stored RBCs may enhance transfusion-related complications, such as infection.

  20. Transfusion of human volunteers with older, stored red blood cells produces extravascular hemolysis and circulating non–transferrin-bound iron

    PubMed Central

    Brittenham, Gary M.; Billote, Genia B.; Francis, Richard O.; Ginzburg, Yelena Z.; Hendrickson, Jeanne E.; Jhang, Jeffrey; Schwartz, Joseph; Sharma, Shruti; Sheth, Sujit; Sireci, Anthony N.; Stephens, Hannah L.; Stotler, Brie A.; Wojczyk, Boguslaw S.; Zimring, James C.; Spitalnik, Steven L.

    2011-01-01

    Transfusions of RBCs stored for longer durations are associated with adverse effects in hospitalized patients. We prospectively studied 14 healthy human volunteers who donated standard leuko-reduced, double RBC units. One unit was autologously transfused “fresh” (3-7 days of storage), and the other “older” unit was transfused after 40 to 42 days of storage. Of the routine laboratory parameters measured at defined times surrounding transfusion, significant differences between fresh and older transfusions were only observed in iron parameters and markers of extravascular hemolysis. Compared with fresh RBCs, mean serum total bilirubin increased by 0.55 mg/dL at 4 hours after transfusion of older RBCs (P = .0003), without significant changes in haptoglobin or lactate dehydrogenase. In addition, only after the older transfusion, transferrin saturation increased progressively over 4 hours to a mean of 64%, and non–transferrin-bound iron appeared, reaching a mean of 3.2μM. The increased concentrations of non–transferrin-bound iron correlated with enhanced proliferation in vitro of a pathogenic strain of Escherichia coli (r = 0.94, P = .002). Therefore, circulating non–transferrin-bound iron derived from rapid clearance of transfused, older stored RBCs may enhance transfusion-related complications, such as infection. The trial was registered with www.clinicaltrials.gov as #NCT01319552. PMID:22021369

  1. Blood pressure and culture. The contribution of cross-cultural comparisons to psychosomatics.

    PubMed

    Murphy, H B

    1982-01-01

    It is well known that mean blood pressure levels tend to be low in non-westernized tribal peoples and that these levels tend to rise, particularly in the older age groups, among persons of the same origins who come into more contact with modern Western life-styles. That tendency can be attributed to many factors - increased salt intake, increased obesity, acculturation anxiety, information overload, increased competitiveness, envious resentment, etc. Disentangling these various hypothesized factors is virtually impossible when studying patients or population samples from a single sociocultural group, but cross-cultural comparisons may under favorable circumstances permit some such disentangling. Using data from Micronesia, Polynesia, and East Africa, an attempt will be made to assess which types of psychological stress are most likely to conduce to hypertension, and how certain traditional cultures may have been reducing these stresses.

  2. Evaluation of store lesion in platelet obtained by apheresis compared to platelet derived from whole blood and its impact on the in vitro functionality.

    PubMed

    Quintero, M; Núñez, M; Mellado, S; Maldonado, M; Wehinger, S

    2015-12-01

    Platelet units for transfusion purposes are obtained manually from whole blood or by apheresis, in an automated process. In both methods, platelets during storage present a characteristics grouped under the name "storage lesion" that are associated with adverse effects on platelet units. Oxidative stress has been claimed to be one of major causes, leading to activation and apoptosis processes affecting their post transfusion functionality. In this work, we observed an association between apheresis and a reduced presence of oxidative stress and better results in functional markers in stored platelets, compared to manually obtained platelets. Then, apheresis which would ensure a greater number of functional platelets during the 5 days of storage, compared to concentrates obtained from whole blood.

  3. Gas-liquid chromatography in routine processing of blood cultures for detecting anaerobic bacteraemia.

    PubMed Central

    Reig, M; Molina, D; Loza, E; Ledesma, M A; Meseguer, M A

    1981-01-01

    Gas-liquid chromatography was performed on 233 positive blood cultures and findings were compared with culture results. Obligate anaerobic bacteria were recovered from 78 out of 79 blood cultures containing butyric or iso-valeric acids, or both; from 28 out of 69 blood cultures containing succinic acid; and from only one out of 41 blood cultures containing succinic but not butyric or iso-valeric acid. Good correlations (88%) were found for the recovery of anaerobic bacteria and the detection of butyric and/or iso-valeric acids. Detecting volatile fatty acids by gas-liquid chromatography performed on blood cultures at the first signs of growth can therefore provide an early and reliable indication of the presence of anaerobic bacteria. PMID:7014645

  4. Neuronal zinc stores are modulated by non-steroidal anti-inflammatory drugs: an optical analysis in cultured hippocampal neurons.

    PubMed

    Love, Rachal; Salazar, Gloria; Faundez, Victor

    2005-11-02

    Zinc chelation and non-steroidal anti-inflammatory drugs (NSAIDs) have been explored as potential neuroprotective agents. However, it remains unknown whether NSAIDs and zinc chelation may converge on a similar cellular process. Using two-photon microscopy to observe hippocampal neurons labeled with a zinc-sensitive dye, we provide evidence that three chemically unrelated NSAIDs, niflumic acid, ibuprofen, and naproxen, acutely increase intracellular zinc stores from extracellular metal pools. Phospholipase A2 inhibitors triggered similar responses, suggesting that NSAIDs likely control zinc stores by their activity as cyclooxygenase inhibitors. These results provide evidence for a new link between cyclooxygenase metabolites and the mechanisms controlling neuronal zinc pools.

  5. Comparison of blood culture and multiplex real-time PCR for the diagnosis of nosocomial sepsis.

    PubMed

    Dinç, Fatih; Akalin, Halis; Özakin, Cüneyt; Sinirtaş, Melda; Kebabçi, Nesrin; Işçimen, Remzi; Kelebek Girgin, Nermin; Kahveci, Ferda

    2016-03-01

    In many cases of suspected sepsis, causative microorganisms cannot be isolated. Multiplex real-time PCR generates results more rapidly than conventional blood culture systems. In this study, we evaluated the diagnostic performance of multiplex real-time PCR (LightCycler® SeptiFast, Roche, Mannheim, Germany), and compared with blood cultures and cultures from focus of infection in nosocomial sepsis. Seventy-eight nosocomial sepsis episodes in 67 adult patients were included in this study. The rates of microorganism detection by blood culture and PCR were 34.2% and 47.9%, respectively. Sixty-five microorganisms were detected by both methods from 78 sepsis episodes. Nineteen of these microorganisms were detected by both blood culture and PCR analysis from the same sepsis episode. There was statistically moderate concordance between the two methods (κ=0.445, P<0.001). There was no significant agreement between the blood culture and PCR analysis in terms of microorganism detected (κ=0.160, P=0.07). Comparison of the results of PCR and cultures from focus of infection revealed no significant agreement (κ=0.110, P=0.176). However, comparison of the results of PCR and blood cultures plus cultures from focus of infection (positive blood culture and/or positive culture from focus of infection) showed poor agreement (κ=0.17, P=0.026). When the blood culture was used as the gold standard, the sensitivity, specificity, positive and negative predictive value of PCR in patients with bacteremia was 80%, 69%, 57% and 87%, respectively. SeptiFast may be useful when added to blood culture in the diagnosis and management of sepsis.

  6. Detection of newly produced T and B lymphocytes by digital PCR in blood stored dry on nylon flocked swabs.

    PubMed

    Tessitore, Marion Vaglio; Sottini, Alessandra; Roccaro, Aldo M; Ghidini, Claudia; Bernardi, Simona; Martellosio, Giovanni; Serana, Federico; Imberti, Luisa

    2017-04-05

    A normal number of T-cell receptor excision circles (TRECs) and K-deleting recombination excision circles (KRECs) is considered a biomarker for adequate new T- and B-cell production. In newborns, detection of TRECs and KRECs by real time PCR from dried blood spotted on filter paper is used for the screening of severe immunodeficiency. In adults, elderly and during diseases, where the number of TRECs is lower than in newborns and children, a large amount of DNA and a sensitive method of amplification are necessary to identify newly produced lymphocytes. DNA was prepared from blood of 203 healthy adults (range: 18-91 years old) absorbed for 10 s on flocked swabs and let to dry, or from peripheral blood mononuclear cells. DNA was subjected to digital PCR and to well established conventional real time PCR-based method using TREC- and KREC-specific primers and probes. The number of TRECs and KRECs was expressed per mL of blood. Statistical analysis was performed by nested ANOVA, Pearson coefficient of determination, and by linear regression tests. The novel method for the storage of dried blood on nylon flocked swabs and the use of digital PCR allow quantification of TRECs and KRECs with high degree of sensitivity, specificity, accuracy, and precision. TRECs and KRECs were amplified by digital PCR in all tested blood samples, including those obtained from elderly individuals (>70 years old) and that were negative by real time PCR. Furthermore, values of TRECs and KRECs obtained by digital PCR were in the range of those acquired by real time PCR. Our findings demonstrate that DNA isolation from dried blood on flocked swabs followed by digital PCR-based analysis represents a useful tool for studying new lymphocyte production in adults and elderly individuals. This suggests the potential use of the methodology when monitoring of clinical variables is limited by the number of molecules that can be amplified and detected, such as in patients with immunodeficiency or under

  7. Reconstituted fresh whole blood improves clinical outcomes compared with stored component blood therapy for neonates undergoing cardiopulmonary bypass for cardiac surgery: a randomized controlled trial.

    PubMed

    Gruenwald, Colleen E; McCrindle, Brian W; Crawford-Lean, Lynn; Holtby, Helen; Parshuram, Christopher; Massicotte, Patricia; Van Arsdell, Glen

    2008-12-01

    This study compared the effects of reconstituted fresh whole blood against standard blood component therapy in neonates undergoing cardiac surgery. Patients less than 1 month of age were randomized to receive either reconstituted fresh whole blood (n = 31) or standard blood component therapy (n = 33) to prime the bypass circuit and for transfusion during the 24 hours after cardiopulmonary bypass. Primary outcome was chest tube drainage; secondary outcomes included transfusion needs, inotrope score, ventilation time, and hospital length of stay. Patients who received reconstituted fresh whole blood had significantly less postoperative chest tube volume loss per kilogram of body weight (7.7 mL/kg vs 11.8 mL/kg; P = .03). Standard blood component therapy was associated with higher inotropic score (6.6 vs 3.3; P = .002), longer ventilation times (164 hours vs 119 hours; P = .04), as well as longer hospital stays (18 days vs 12 days; P = .006) than patients receiving reconstituted fresh whole blood. Of the different factors associated with the use of reconstituted fresh whole blood, lower platelet counts at 10 minutes and at the end of cardiopulmonary bypass, older age of cells used in the prime and throughout bypass, and exposures to higher number of allogeneic donors were found to be independent predictors of poor clinical outcomes. Reconstituted fresh whole blood used for the prime, throughout cardiopulmonary bypass, and for all transfusion requirements within the first 24 hours postoperatively results in reduced chest tube volume loss and improved clinical outcomes in neonatal patients undergoing cardiac surgery.

  8. Low yield of blood and wound cultures in patients with skin and soft-tissue infections.

    PubMed

    Torres, Jesus; Avalos, Nathaniel; Echols, Lamarr; Mongelluzzo, Jillian; Rodriguez, Robert M

    2017-08-01

    Current guidelines recommend blood cultures in skin and soft-tissue infection (SSTI) patients only with signs of systemic toxicity and wound cultures for severe purulent infections. Our objectives were to determine: 1) blood and wound culture yields in patients admitted with SSTIs; 2) whether injection drug users (IDUs) and febrile patients had higher blood culture yields; and 3) whether blood and wound cultures grew organisms sensitive to typical SSTI empiric antibiotics. We prospectively enrolled adult patients admitted from the ED with SSTIs at an urban hospital. We recorded patient characteristics, including IDU, comorbidities and temperatures, and followed admitted patients throughout their hospital course. Of 734 SSTI patients enrolled, 246 (33.5%) were admitted. Of 86 (35.0%) patients who had blood cultures, six had positive cultures (yield=7.0%; 95% confidence intervals [CIs] 3.2-14.4); 4 were methicillin sensitive Staphylococcus aureus (MSSA) and 2 were methicillin resistant (MRSA). Of 29 febrile patients, 1 had a positive culture (yield=3.5%; 95% CI 0.6-17.2). Of 101 admitted IDU patients, 46 (46%) received blood cultures, and 4 had positive cultures (yield=8.7%; 95% CI 3.4-20.3). Of 89 patients with purulent wounds, 44 (49.4%) patients had ED wound cultures. Thirteen had positive cultures (yield=29.6%; 95% CI 18.2-44.2%). Most were MRSA, MSSA, and group A Streptococcus species - all sensitive to Vancomycin. Febrile and IDU patients had low yields of blood cultures similar to yields in non-IDU and afebrile patients. All blood and wound culture species were adequately covered by currently recommended empiric antibiotic regimens. Copyright © 2017 Elsevier Inc. All rights reserved.

  9. Effect of sodium polyanetholesulfonate and gelatin on the recovery of Gardnerella vaginalis from blood culture media.

    PubMed Central

    Reimer, L G; Reller, L B

    1985-01-01

    Sodium polyanetholesulfonate (SPS) is used as a routine supplement to blood culture media to enhance recovery of microorganisms, but it inhibits the growth of Peptostreptococcus anaerobius, Neisseria meningitidis, Neisseria gonorrhoeae, and Streptobacillus moniliformis. Comparative clinical blood culture studies at the University of Colorado Hospital suggested that SPS also inhibits the growth of Gardnerella vaginalis. We inoculated 16 blood culture isolates of G. vaginalis into 11 blood culture media containing SPS or sodium amylosulfate, with and without gelatin. In the absence of gelatin, only brain heart infusion and thiol broths with SPS supported the growth of more than five strains of G. vaginalis, whereas all media except Bactec 6B and 7C and brucella broths recovered most isolates with SPS and gelatin or with sodium amylosulfate alone. We conclude that SPS inhibits the growth of G. vaginalis in blood culture media but that this inhibition is medium dependent and can be overcome by supplementation of most media with gelatin. PMID:2987298

  10. Evaluation of an intervention to improve blood culture practices: a cluster randomised trial.

    PubMed

    Pavese, P; Maillet, M; Vitrat-Hincky, V; Recule, C; Vittoz, J-P; Guyomard, A; Seigneurin, A; François, P

    2014-12-01

    This study aimed to evaluate an intervention to improve blood culture practices. A cluster randomised trial in two parallel groups was performed at the Grenoble University Hospital, France. In October 2009, the results of a practices audit and the guidelines for the optimal use of blood cultures were disseminated to clinical departments. We compared two types of information dissemination: simple presentation or presentation associated with an infectious diseases (ID) specialist intervention. The principal endpoint was blood culture performance measured by the rate of patients having one positive blood culture and the rate of positive blood cultures. The cases of 130 patients in the "ID" group and 119 patients in the "simple presentation" group were audited during the second audit in April 2010. The rate of patients with one positive blood culture increased in both groups (13.62 % vs 9.89 % for the ID group, p = 0.002, 15.90 % vs 13.47 % for the simple presentation group, p = 0.009). The rate of positive blood cultures improved in both groups (6.68 % vs 5.96 % for the ID group, p = 0.003, 6.52 % vs 6.21 % for the simple presentation group, p = 0.017). The blood culture indication was significantly less often specified in the request form in the simple presentation group, while it remained stable in the ID group (p = 0.04). The rate of positive blood cultures and the rate of patients having one positive blood culture improved in both groups. The ID specialist intervention did not have more of an impact on practices than a simple presentation of audit feedback and guidelines.

  11. Routine blood cultures in the management of pyelonephritis in pregnancy for improving outcomes.

    PubMed

    Gomi, Harumi; Goto, Yoshihito; Laopaiboon, Malinee; Usui, Rie; Mori, Rintaro

    2015-02-13

    Pyelonephritis is a type of urinary tract infection (UTI) that affects the upper urinary tract and kidneys, and is one of the most common conditions for hospitalisation among pregnant women, aside from delivery. Samples of urine and blood are obtained and used for cultures as part of the diagnosis and management of the condition. Acute pyelonephritis requires hospitalisation with intravenous administration of antimicrobial agents. Several studies have questioned the necessity of obtaining blood cultures in addition to urine cultures, citing cost and questioning whether blood testing is superfluous. Pregnant women with bacteraemia require a change in the initial empirical treatment based on the blood culture. However, these cases are not common, and represent approximately 15% to 20% of cases. It is unclear whether blood cultures are essential for the effective management of the condition. To assess the effectiveness of routine blood cultures to improve health outcomes in the management of pyelonephritis in pregnant women. We searched the Cochrane Pregnancy and Childbirth Group's Trials Register without language or date restrictions (31 December 2014). Randomised controlled trials and quasi-randomised trials comparing outcomes among pregnant women with pyelonephritis who received initial management with or without blood cultures. Cluster-randomised trials were eligible for inclusion in this review but none were identified. Clinical trials using a cross-over design were not eligible for inclusion. Two review authors independently assessed one trial report for inclusion. We identified one trial report but this was excluded. No clinical trials met the inclusion criteria for this review. There are no large-scale randomised controlled trials to assess outcomes in the management of pyelonephritis in pregnancy with or without blood cultures. Randomised controlled trials are needed to evaluate the effectiveness of managing pyelonephritis in pregnant women with or without

  12. Clinical evaluation of the FilmArray blood culture identification panel in identification of bacteria and yeasts from positive blood culture bottles.

    PubMed

    Altun, Osman; Almuhayawi, Mohammed; Ullberg, Måns; Ozenci, Volkan

    2013-12-01

    The FilmArray platform (FA; BioFire, Salt Lake City, UT) is a closed diagnostic system allowing high-order multiplex PCR analysis with automated readout of results directly from positive blood cultures in 1 h. In the present study, we evaluated the clinical performance of the FilmArray blood culture identification (BCID) panel, which includes 19 bacteria, five yeasts, and three antibiotic resistance genes. In total, 206 blood culture bottles were included in the study. The FilmArray could identify microorganisms in 153/167 (91.6%) samples with monomicrobial growth. Thirteen of the 167 (7.8%) microorganisms were not covered by the FilmArray BCID panel. In 6/167 (3.6%) samples, the FilmArray detected an additional microorganism compared to blood culture. When polymicrobial growth was analyzed, the FilmArray could detect all target microorganisms in 17/24 (71%) samples. Twelve blood culture bottles that yielded a positive signal but showed no growth were also negative by FilmArray. In 3/206 (1.5%) bottles, the FilmArray results were invalid. The results of the FilmArray were reproducible, as demonstrated by the testing and retesting of five bottles in the same day and a longitudinal follow-up of five other blood cultures up to 4 weeks. The present study shows that the FilmArray is a rapid identification method with high performance in direct identification of bacteria and yeasts from positive blood culture bottles.

  13. Diagnostic assays for identification of microorganisms and antimicrobial resistance determinants directly from positive blood culture broth.

    PubMed

    Pence, Morgan A; McElvania TeKippe, Erin; Burnham, Carey-Ann D

    2013-09-01

    The detection of blood stream infections is one of the most important functions of the clinical microbiology laboratory. Sepsis is a clinical emergency, and mortality increases if commencement of appropriate antimicrobial therapy is delayed. Automated blood culture systems are the most sensitive approach for detection of the causative agent of sepsis. Several laboratory methods have been developed to expedite identification of organisms directly from positive blood culture broth. The principle and analytical performance characteristics of these methods are described in this review.

  14. Automated preparation of whole blood-derived platelets suspended in two different platelet additive solutions and stored for 7 days.

    PubMed

    Cid, Joan; Magnano, Laura; Molina, Patricia; Diaz-Ricart, Maribel; Martínez, Nuria; Maymó, Rosa M; Puig, Lluís; Lozano, Miguel; Escolar, Ginés; Galán, Ana M

    2014-02-01

    The Atreus system (Terumo BCT) automates the preparation of blood components from whole blood donations. Intermediate platelet (PLT) products can be pooled manually or with the OrbiSac (Terumo BCT) and suspended in different PLT additive solutions (PASs) to obtain PLT concentrates (PCs). The aim of our study was to compare the in vitro PLT quality of PCs obtained with either the Atreus 2C+ and the OrbiSac or the Atreus 3C and suspended in PAS-II or PAS-IIIM during storage for up to 7 days. We prepared eight PCs from buffy coats obtained with Atreus 2C+, pooled with the OrbiSac, and suspended in PAS-II and eight PCs from interim PLT units obtained with the Atreus 3C and suspended either in PAS-II or in PAS-IIIM. We measured volume, PLT content, and mean PLT component and performed metabolic assays (pH, glucose, lactate, pO₂, and pCO₂) and flow cytometry analyses (GPIb, GPIIbIIIa, GPIV, CD62P, CD63, von Willebrand factor [vWF], fibrinogen, Factor V, and annexin V). PCs prepared with the Atreus 3C showed lower volume and higher PLT concentration when compared with PCs prepared with the Atreus 2C+ and the OrbiSac (p < 0.05). Glucose consumption rate and the expression of CD62P, CD63, and vWF were lower in PCs suspended in PAS-IIIM when compared with PCs suspended in PAS-II (p < 0.05). PCs prepared with the Atreus 3C and suspended in PAS-IIIM preserve satisfactorily the in vitro PLT quality during 7-day storage. PLT activation during a 7-day storage period was lower when the storage solution was PAS-IIIM in comparison with PAS-II. © 2013 American Association of Blood Banks.

  15. Intrabone Transplant of Cord Blood Stem Cells Establishes a Local Engraftment Store: A Functional PET/FDG Study

    PubMed Central

    Marini, Cecilia; Podestà, Marina; Massollo, Michela; Capitanio, Selene; Fiz, Francesco; Morbelli, Silvia; Brignone, Massimo; Bacigalupo, Andrea; Piana, Michele; Frassoni, Francesco; Sambuceti, Gianmario

    2012-01-01

    Background. Despite advancements in comprehension of molecular mechanisms governing bone marrow (BM) homing of hematopoietic stem cells, cord blood transplant (CBT) suffers from a slow rate of hematopoietic recovery. Intrabone (IB) injection has been proposed as a method able to improve speed of BM engraftment with respect to conventional IV protocols. However, the mechanisms underlying this benefit are largely unknown. Aim. To verify whether IB-CBT determines a local engraftment able to predict the reconstitution of recipient hematopoiesis. Design and Methods. Twenty-one patients with hematologic malignancies received IB injection into both iliac crests of 3.2 ± 0.68 ∗ 107/kg cord blood cells. One month following IB-CBT, PET-CT imaging was performed. Maximal standardized uptake values (SUVs) were assessed in BM of both iliac crests and in all lumbar vertebrae. Results. Maximal SUV within iliac crests was higher than in lumbar vertebrae (4.1 ± 1.7 versus 3.2 ± 0.7, resp., P = 0.01). However, metabolic activity in these two different BM districts was significantly correlated (r = 0.7, P < 0.001). Moreover, FDG uptake values within the injection site closely predicted platelet recovery 100 days after IB-CBT (r = 0.72, P < 0.01). Conclusions. The metabolic activity of injected BM predicts the subsequent rate of hematopoietic recovery after IB-CBT, suggesting a pivotal role of the local engraftment in the reconstitution of recipient hematopoiesis. PMID:23093864

  16. Storing acorns

    Treesearch

    Kristina. Connor

    2004-01-01

    We examined changes that occurred in acorns during storage at different temperatures and moisture contents over a period of 3 y. In general, we found that to achieve optimum viability, acorns must be stored fully hydrated. Acorns also survived longer and sprouted less while in storage if stored at –2 °C (28 °F) instead of the usual 4 °C (39 °F). However, we suspect...

  17. Long-term stability of morphine, codeine, and 6-acetylmorphine in real-life whole blood samples, stored at -20°C.

    PubMed

    Høiseth, Gudrun; Fjeld, Bente; Burns, Margrete Larsen; Strand, Dag Helge; Vindenes, Vigdis

    2014-06-01

    Stability of drugs during storage is important in forensic toxicology. For the analytes detected after intake of heroin (6-acetylmorphine (6-AM), morphine and codeine), long-time stability in real life whole blood samples are studied in only a small number of cases. Whole blood post mortem (n=37) and whole blood samples from living persons (n=22) containing morphine and codeine as well as 6-AM in blood or urine were selected. All cases represented intake of heroin. All samples contained fluoride and were initially analysed and stored in normal conditions (-20°C) for 4-9 years. All samples were then reanalysed using the same analytical methods and the results were compared. For samples from living persons, the median change in concentration was -3.7% for morphine and -5.3% for codeine. For post mortem samples, the median change in concentration was -12% for morphine and -11% for codeine. Both for samples from living persons and post mortem samples, the decrease in the concentrations from the original analysis to reanalysis were statistically significant for morphine and codeine. Regarding 6-AM, all living samples were negative at reanalysis. For post mortem samples, four cases still tested positive for 6-AM at reanalysis with a median change in the concentrations of -81%. There was no significant change in the morphine to codeine concentration ratios neither for living nor post mortem samples. This study showed that in real life whole blood samples, the concentrations of morphine and codeine are relatively stable during long-term storage at -20°C. 6-AM on the other hand, shows a considerable decrease in concentrations that is important to consider when interpreting results from reanalyses of forensic cases. Copyright © 2014 Elsevier Ireland Ltd. All rights reserved.

  18. The relationship between body iron stores and blood and urine cadmium concentrations in US never-smoking, non-pregnant women aged 20-49 years

    SciTech Connect

    Gallagher, Carolyn M.; Chen, John J.; Kovach, John S.

    2011-07-15

    Background: Cadmium is a ubiquitous environmental pollutant associated with increased risk of leading causes of mortality and morbidity in women, including breast cancer and osteoporosis. Iron deficiency increases absorption of dietary cadmium, rendering women, who tend to have lower iron stores than men, more susceptible to cadmium uptake. We used body iron, a measure that incorporates both serum ferritin and soluble transferrin receptor, as recommended by the World Health Organization, to evaluate the relationships between iron status and urine and blood cadmium. Methods: Serum ferritin, soluble transferrin receptor, urine and blood cadmium values in never-smoking, non-pregnant, non-lactating, non-menopausal women aged 20-49 years (n=599) were obtained from the 2003-2008 National Health and Nutrition Examination Surveys. Body iron was calculated from serum ferritin and soluble transferrin receptor, and iron deficiency defined as body iron <0 mg/kg. Robust linear regression was used to evaluate the relationships between body iron and blood and urine cadmium, adjusted for age, race, poverty, body mass index, and parity. Results: Per incremental (mg/kg) increase in body iron, urine cadmium decreased by 0.003 {mu}g/g creatinine and blood cadmium decreased by 0.014 {mu}g/L. Iron deficiency was associated with 0.044 {mu}g/g creatinine greater urine cadmium (95% CI=0.020, 0.069) and 0.162 {mu}g/L greater blood cadmium (95% CI=0.132, 0.193). Conclusions: Iron deficiency is a risk factor for increased blood and urine cadmium among never-smoking, pre-menopausal, non-pregnant US women, independent of age, race, poverty, body mass index and parity. Expanding programs to detect and correct iron deficiency among non-pregnant women merits consideration as a potential means to reduce the risk of cadmium associated diseases. - Highlights: {yields} Body iron was calculated from serum ferritin and soluble transferrin receptor. {yields} Body iron was inversely associated with blood

  19. Do emergency department blood cultures change practice in patients with pneumonia?

    PubMed

    Kennedy, Maura; Bates, David W; Wright, Sharon B; Ruiz, Raul; Wolfe, Richard E; Shapiro, Nathan I

    2005-11-01

    Although it is considered standard of care to obtain blood cultures on patients hospitalized for pneumonia, several studies have questioned the utility and cost-effectiveness of this practice. The objective of this study is to determine the impact of emergency department (ED) blood cultures on antimicrobial therapy for patients with pneumonia. We performed a prospective, observational, cohort study of consecutive adult (age > or =18 years) patients treated at an urban university ED between February 1, 2000 and February 1, 2001. Inclusion criteria were radiographic evidence of pneumonia, clinical evidence of pneumonia, and blood culture obtained. Blood cultures were classified as positive, negative, or contaminant based on previously established criteria. Additionally, data were collected on antimicrobial sensitivities, empiric antibiotic therapy, antibiotic changes, and reasons for changes. There were 3,926 ED visits with blood cultures obtained for any reason, of which 3,762 (96%) were available for review. Of these, 414 of 3,762 (11%) patients met pneumonia study inclusion criteria, and blood cultures identified 29 of 414 (7.0%) patients with true bacteremia. In the 414 patients, blood culture results altered therapy for 15 patients (3.6%) with suspected pneumonia, of which 11 (2.7%) patients had their coverage narrowed; only 4 (1.0%) patients had their coverage broadened because of resistance to empiric therapy. For the 11 patients with bacteremia whose therapy was not altered, culture results actually supported narrowing therapy in 8 (1.9%) cases, but this was not done. Blood cultures rarely altered therapy for patients presenting to the ED with pneumonia. More discriminatory blood culture use may potentially reduce resource utilization.

  20. Potential Impact of Rapid Blood Culture Testing for Gram-Positive Bacteremia in Japan with the Verigene Gram-Positive Blood Culture Test

    PubMed Central

    Matsuda, Mari; Iguchi, Shigekazu; Mizutani, Tomonori; Hiramatsu, Keiichi; Tega-Ishii, Michiru; Sansaka, Kaori; Negishi, Kenta; Shimada, Kimie; Umemura, Jun; Notake, Shigeyuki; Yanagisawa, Hideji; Yabusaki, Reiko; Araoka, Hideki; Yoneyama, Akiko

    2017-01-01

    Background. Early detection of Gram-positive bacteremia and timely appropriate antimicrobial therapy are required for decreasing patient mortality. The purpose of our study was to evaluate the performance of the Verigene Gram-positive blood culture assay (BC-GP) in two special healthcare settings and determine the potential impact of rapid blood culture testing for Gram-positive bacteremia within the Japanese healthcare delivery system. Furthermore, the study included simulated blood cultures, which included a library of well-characterized methicillin-resistant Staphylococcus aureus (MRSA) and vancomycin-resistant enterococci (VRE) isolates reflecting different geographical regions in Japan. Methods. A total 347 BC-GP assays were performed on clinical and simulated blood cultures. BC-GP results were compared to results obtained by reference methods for genus/species identification and detection of resistance genes using molecular and MALDI-TOF MS methodologies. Results. For identification and detection of resistance genes at two clinical sites and simulated blood cultures, overall concordance of BC-GP with reference methods was 327/347 (94%). The time for identification and antimicrobial resistance detection by BC-GP was significantly shorter compared to routine testing especially at the cardiology hospital, which does not offer clinical microbiology services on weekends and holidays. Conclusion. BC-GP generated accurate identification and detection of resistance markers compared with routine laboratory methods for Gram-positive organisms in specialized clinical settings providing more rapid results than current routine testing. PMID:28316631

  1. Assessment of apoptosis, MMP-1, MMP-3, TIMP-2 expression and mechanical and biochemical properties of fresh rabbit's medial meniscus stored two weeks under tissue culture conditions

    PubMed Central

    Stasikowska-Kanicka, Olga; Janus, Jolanta; Konecki, Włodzimierz; Danilewicz, Marian; Fabiś, Jarosław

    2014-01-01

    Introduction The purpose of this study was to assess apoptosis, the expression of MMP-1, MMP-3 and TIMP-2, as well as the mechanical and biochemical properties of fresh rabbit medial meniscus after 2 weeks stored under tissue culture conditions. Material and methods The study material included 26 rabbit's medial menisci. Fourteen menisci were stored for 2 weeks under tissue culture conditions. Thirteen menisci were subjected to immunohistochemistry tests. Apoptosis (TUNEL method) and the expression of MMP-1 (collagenase-1), MMP-3 (stromelysin-2) and TIMP-2 (tissue inhibitor of metalloproteinases-2) were estimated semiquantitatively. The remaining menisci were tested mechanically and biochemically. The mechanical properties were assessed by the degree of elasticity. Biochemical composition was based on the content analysis of water, total collagen and glycosaminoglycans. Results As in the control group, the conducted study did not reveal any apoptosis in the stored menisci. The study group showed a slight expression of MMP-1 (0.27 ±0.19), MMP-3 (0.30 ±0.20) and TIMP-2 (0.33 ±0.20) – no statistically significant difference from controls. The degree of elasticity was 28.51 ±1.53 in the study group, and this did not statistically differ from the control group. No significant biochemical differences were identified in any other monitored parameter. Conclusions The conducted in vitro study did not show any negative influence of a 2-week storage period under tissue culture conditions on the apoptosis and expression of MMP-1, MMP-3 and TIMP-2 in rabbit fresh menisci, nor any impact on their mechanical and biochemical properties. PMID:24701230

  2. Repeated Blood Cultures in Pediatric Febrile Neutropenia: Would Following the Guidelines Alter the Outcome?

    PubMed

    Petty, Lindsay A; Sokol, Elizabeth A; Bartlett, Allison H; McNeer, Jennifer L; Alexander, Kenneth A; Pisano, Jennifer

    2016-07-01

    The Infectious Diseases Society of America (IDSA) guidelines recommend collecting blood cultures for the first 3 days of febrile neutropenia (FN) in the clinically stable oncology patient with persistent fevers. Nonetheless, many physicians send daily blood cultures beyond 3 days, and the impact of that practice is uncertain. We reviewed pediatric FN episodes from July 2009 to May 2014 at University of Chicago Comer Children's Hospital. For each positive culture, we determined if it was a pathogen or a contaminant. We reviewed episode and patient demographics to identify risk factors for subsequent positive blood cultures in the setting of an initially negative culture. We identified 381 episodes of FN in 162 patients. Of those, 87 had a positive blood culture on day 1 (21.0% incidence of bacteremia). Of 294 episodes with a negative blood culture on day 1, six (2.04%, 95% confidence interval [CI] 0.42-3.67) had a positive culture after day 3. Of those, three were pathogens (1.02%, 95%CI -0.14 to 2.18), and only one was found in a hemodynamically stable patient (0.34%, 95%CI -0.33 to 1.01) with new mucositis. In the other two patients, Escherichia coli was isolated from blood cultures after day 10 in the setting of significant hemodynamic changes. Risk factor analysis performed in stable patients yielded nonsignificant results. Of 294 FN episodes with an initial negative blood culture, only one episode of bacteremia occurred without hemodynamic changes past day 3, supporting the IDSA guidelines to discontinue blood cultures in stable FN patients after day 3. © 2016 Wiley Periodicals, Inc.

  3. Growth of nutritionally variant streptococci on common laboratory and 10 commercial blood culture media.

    PubMed Central

    Reimer, L G; Reller, L B

    1981-01-01

    Nutritionally variant streptococci fail to grow on routine sheep blood agar plates. Moreover, these strains are a recognized cause of culture-negative endocarditis. We tested the ability of chocolate and brucella blood agars, sheep blood agar with a staphylococcal streak, sheep blood agar with 0.001% pyridoxal, and 10 commercial blood culture media from two manufactures to grow these bacteria. Of the original 25 strains tested, 16 were recovered on chocolate agar, 21 were recovered on brucella blood agar and 21 were recovered on sheep blood agar with a staphylococcal streak. Sheep blood agar with pyridoxal grew all 22 strains tested. Supplemented peptone, thioglycolate, and thiol broths grew all strains, but brain heart infusion and three tryptic soy broths supported five or fewer strains. The addition of 5 ml of human blood improved recovery to 100% in all media except tryptic soy broths. Unless supplemented wih pyridoxal, common laboratory agars were inadequate for recovering all strains of variant streptococci upon subculture of blood culture bottles. As used clinically, the blood culture media that we studied other than tryptic soy broths should reliably grow these bacteria. PMID:7287889

  4. Comparative evaluation of paired blood culture (aerobic/aerobic) and single blood culture, along with clinical importance in catheter versus peripheral line at a tertiary care hospital.

    PubMed

    Tarai, B; Das, P; Kumar, D; Budhiraja, S

    2012-01-01

    Paired blood culture (PBC) is uncommon practice in hospitals in India, leading to delayed and inadequate diagnosis. Also contamination remains a critical determinant in hampering the definitive diagnosis. To establish the need of PBC over single blood culture (SBC) along with the degree of contamination, this comparative retrospective study was initiated. We processed 2553 PBC and 4350 SBC in BacT/ALERT 3D (bioMerieux) between October 2010 and June 2011. The positive cultures were identified in VITEK 2 Compact (bioMerieux). True positivity and contaminants were also analyzed in 486 samples received from catheter and peripheral line. Out of 2553 PBC samples, positivity was seen in 350 (13.70%). In 4350 SBC samples, positivity was seen in 200 samples (4.59%). In PBC true pathogens were 267 (10.45%) and contaminants were 83 (3.25%), whereas in SBC 153 (3.51%) were true positives and contaminants were 47 (1.08%). Most of the blood cultures (99.27 %) grew within 72 h and 95.8% were isolated within 48 h. In 486 PBCs received from catheter/periphery (one each), catheter positivity was found in 85 (true positives were 48, false positives 37). In peripheral samples true positives were 50 and false positives were 8. Significantly higher positive rates were seen in PBCs compared with SBCs. Automated blood culture and identification methods significantly reduced the time required for processing of samples and also facilitated yield of diverse/rare organisms. Blood culture from catheter line had higher false positives than peripheral blood culture. Thus every positive result from a catheter must be correlated with clinical findings and requires further confirmation.

  5. Comparative evaluation of the role of single and multiple blood specimens in the outcome of blood cultures using BacT/ALERT 3D (automated) blood culture system in a tertiary care hospital

    PubMed Central

    Elantamilan, D.; Lyngdoh, Valarie Wihiwot; Khyriem, Annie B.; Rajbongshi, Jyotismita; Bora, Ishani; Devi, Surbala Thingujam; Bhattacharyya, Prithwis; Barman, Himesh

    2016-01-01

    Introduction: Bloodstream infection (BSI) is a leading cause of mortality in critically ill patients. The mortality directly attributable to BSI has been estimated to be around 16% and 40% in general hospital population and Intensive Care Unit (ICU) population, respectively. The detection rate of these infections increases with the number of blood samples obtained for culture. The newer continuous monitoring automated blood culture systems with enhanced culture media show increased yield and sensitivity. Hence, we aimed at studying the role of single and multiple blood specimens from different sites at the same time in the outcome of automated blood culture system. Materials and Methods and Results: A total of 1054 blood culture sets were analyzed over 1 year, the sensitivity of one, two, and three samples in a set was found to be 85.67%, 96.59%, and 100%, respectively, which showed a statistically significant difference (P < 0.0001). Similar findings were seen in few more studies, however, among individual organisms in contrast to other studies, the isolation rates of Gram-positive bacteria were less than that of Gram-negative Bacilli with one (or first) sample in a blood culture set. In our study, despite using BacT/ALERT three-dimensional continuous culture monitoring system with FAN plus culture bottles, 15% of positive cultures would have been missed if only a single sample was collected in a blood culture set. Conclusion: The variables like the volume of blood and number of samples collected from different sites still play a major role in the outcome of these automated blood culture systems. PMID:27688629

  6. Factors associated with positive blood cultures in outpatients with suspected bacteremia.

    PubMed

    Wildi, K; Tschudin-Sutter, S; Dell-Kuster, S; Frei, R; Bucher, H C; Nüesch, R

    2011-12-01

    Blood cultures are routinely taken in outpatients with fever and suspected bacterial infections. However, in the majority of cases, they are not informative and of limited value for clinical decision making. The aim of this study was therefore to investigate factors associated with positive blood cultures in outpatients presenting to an outpatient clinic and emergency room. This was a case-control study of all outpatients with positive blood cultures from January 1, 2006 to October 31, 2007 and matched control patients with negative blood cultures in the same time period. Microbiology results and medical charts were reviewed to determine factors associated with positive blood cultures. The presence of a systemic inflammation response syndrome (SIRS) (OR 2.7, 95% Cl 1.0-7.2) and increased C-reactive protein (CRP) (OR 1.1 per 10 mg/l, 95% Cl 1.0-1.2) were the most powerful predictive values for the development of positive blood cultures. In positive cases serum albumin was lower (35 mg/l versus 39 mg/l) than in controls. SIRS, increasing CRP and low albumin were associated with positive blood cultures in outpatients. With simple clinical assessment and few laboratory tests indicative of infection, it is possible to define a group at higher risk for bacteremia in outpatients.

  7. Comparison of PCR, Wright agglutination test and blood culture for diagnosis of brucellosis in suspected patients.

    PubMed

    Hekmatimoghaddam, Seyedhosssein; Sadeh, Maryam; Khalili, Mohammad Bagher; Mollaabedin, Mansour; Sazmand, Alireza

    2013-11-15

    Brucellosis has long been prevalent in Iran, with considerable medical and economic importance. Timely diagnosis is needed for early management and effective prevention of its consequences in human beings and animals. Current diagnostic methods impose peculiar challenges in terms of analytical method performance. This study compares diagnostic sensitivity, specificity, predictive Value of Positive (PVP) and Predictive Value of Negative (PVN) for Polymerase Chain Reaction (PCR), Wright agglutination test and blood culture used for patients suspected of brucellosis. In 120 patients clinically suspected of brucellosis and referred by physicians to the Yazd central Medical Laboratory, some relevant demographic, occupational, nutritional and clinical data were collected. Also, venous blood samples were drawn for diagnosis of brucellosis using PCR, Wright agglutination test and blood culture techniques. The most frequent symptom of patients was arthralgia (82 cases, 68.3%). PCR was positive in 25 cases (20.8%), wright test in 21 patients (17.5%) and blood culture in 6 cases (5%). In 20 out of 21 wright-positive cases, PCR was positive and all of the culture-positive patients had positive PCR. Sensitivity, specificity, PVP and PVN of blood culture compared to PCR (as the gold standard test) were 24, 100, 100 and 86%, respectively, but the above parameters when PCR is compared with blood culture (as gold standard) were 100, 83, 24 and 95%, respectively. PCR has better analytical performances than blood culture for diagnosis of brucellosis and is suitable for confirmation of Wright-positive cases.

  8. What are the critical steps in processing blood cultures? A prospective audit evaluating current practice of reporting blood cultures in a centralised laboratory serving secondary care hospitals.

    PubMed

    Meda, Manjula; Clayton, James; Varghese, Reela; Rangaiah, Jayakeerthi; Grundy, Clive; Dashti, Farnaz; Garner, David; Groves, Katherine; Fitzmaurice, Karen; Hutley, E

    2017-04-01

    To assess current procedures of processing positive blood cultures against national standards with an aim to evaluate its clinical impact and to determine the utility of currently available rapid identification and susceptibility tests in processing of blood cultures. Blood cultures from three secondary care hospitals, processed at a centralised laboratory, were prospectively audited. Data regarding processing times, communication with prescribers, changes to patient management and mortality within 30 days of a significant blood culture were collected in a preplanned pro forma for a 4-week period. Of 2206 blood cultures, 211 positive blood cultures flagged positive. Sixty-nine (3.1%) of all cultures were considered to be contaminated. Fifty per cent of blood cultures that flagged positive had a Gram stain reported within 2 hours. Two (0.99%) patients with a significant bacteraemia had escalation of antimicrobial treatment at the point of reporting the Gram stain that was subsequently deemed necessary once sensitivity results were known. Most common intervention was de-escalation of therapy for Gram-positive organisms at the point of availability of pathogen identification (25.6% in Gram positive vs 10% in Gram negative; p=0.012). For Gram-negative organisms, the most common intervention was de-escalation of therapy at the point of availability of sensitivity results (43% in Gram negatives vs 17.9% in Gram positive; p=0.0097). Overall mortality within 30 days of a positive blood culture was 10.9% (23/211). Antibiotic resistance may have contributed to mortality in four of these patients (three Gram negative and one Gram positive). Gram stain result had the least impact on antibiotic treatment interventions (escalation or de-escalation). Tests that improve identification time for Gram-positive pathogens and sensitivity time for Gram-negative pathogens had the greatest impact in making significant changes to antimicrobial treatment. Published by the BMJ

  9. Low iron stores are related to higher blood concentrations of manganese, cobalt and cadmium in non-smoking, Norwegian women in the HUNT 2 study

    SciTech Connect

    Margrete Meltzer, Helle; Lise Brantsaeter, Anne; Borch-Iohnsen, Berit; Ellingsen, Dag G.; Alexander, Jan; Thomassen, Yngvar; Stigum, Hein; Ydersbond, Trond A.

    2010-07-15

    Low iron (Fe) stores may influence absorption or transport of divalent metals in blood. To obtain more knowledge about such associations, the divalent metal ions cadmium (Cd), manganese (Mn), cobalt (Co), copper (Cu), zinc (Zn) and lead (Pb) and parameters of Fe metabolism (serum ferritin, haemoglobin (Hb) and transferrin) were investigated in 448 healthy, menstruating non-smoking women, age 20-55 years (mean 38 years), participating in the Norwegian HUNT 2 study. The study population was stratified for serum ferritin: 257 were iron-depleted (serum ferritin <12 {mu}g/L) and 84 had iron deficiency anaemia (serum ferritin <12 {mu}g/L and Hb<120 g/L). The low ferritin group had increased blood concentrations of Mn, Co and Cd but normal concentrations of Cu, Zn and Pb. In multiple regression models, ferritin emerged as the main determinant of Mn, Co and Cd (p<0.001), while no significant associations with Cu, Zn and Pb were found. Adjusted r{sup 2} for the models were 0.28, 0.48 and 0.34, respectively. Strong positive associations between blood concentrations of Mn, Co and Cd were observed, also when controlled for their common association with ferritin. Apart from these associations, the models showed no significant interactions between the six divalent metals studied. Very mild anaemia (110{<=}Hb<120 g/L) did not seem to have any effect independent of low ferritin. Approximately 26% of the women with iron deficiency anaemia had high concentrations of all of Mn, Co and Cd as opposed to 2.3% of iron-replete subjects. The results confirm that low serum ferritin may have an impact on body kinetics of certain divalent metal ions, but not all. Only a fraction of women with low iron status exhibited an increased blood concentration of divalent metals, providing indication of complexities in the body's handling of these metals.

  10. Low iron stores are related to higher blood concentrations of manganese, cobalt and cadmium in non-smoking, Norwegian women in the HUNT 2 study.

    PubMed

    Meltzer, Helle Margrete; Brantsaeter, Anne Lise; Borch-Iohnsen, Berit; Ellingsen, Dag G; Alexander, Jan; Thomassen, Yngvar; Stigum, Hein; Ydersbond, Trond A

    2010-07-01

    Low iron (Fe) stores may influence absorption or transport of divalent metals in blood. To obtain more knowledge about such associations, the divalent metal ions cadmium (Cd), manganese (Mn), cobalt (Co), copper (Cu), zinc (Zn) and lead (Pb) and parameters of Fe metabolism (serum ferritin, haemoglobin (Hb) and transferrin) were investigated in 448 healthy, menstruating non-smoking women, age 20-55 years (mean 38 years), participating in the Norwegian HUNT 2 study. The study population was stratified for serum ferritin: 257 were iron-depleted (serum ferritin < 12 microg/L) and 84 had iron deficiency anaemia (serum ferritin < 12 microg/L and Hb < 120 g/L). The low ferritin group had increased blood concentrations of Mn, Co and Cd but normal concentrations of Cu, Zn and Pb. In multiple regression models, ferritin emerged as the main determinant of Mn, Co and Cd (p < 0.001), while no significant associations with Cu, Zn and Pb were found. Adjusted r(2) for the models were 0.28, 0.48 and 0.34, respectively. Strong positive associations between blood concentrations of Mn, Co and Cd were observed, also when controlled for their common association with ferritin. Apart from these associations, the models showed no significant interactions between the six divalent metals studied. Very mild anaemia (110 < or = Hb < 120 g/L) did not seem to have any effect independent of low ferritin. Approximately 26% of the women with iron deficiency anaemia had high concentrations of all of Mn, Co and Cd as opposed to 2.3% of iron-replete subjects. The results confirm that low serum ferritin may have an impact on body kinetics of certain divalent metal ions, but not all. Only a fraction of women with low iron status exhibited an increased blood concentration of divalent metals, providing indication of complexities in the body's handling of these metals. 2010 Elsevier Inc. All rights reserved.

  11. Mirasol Pathogen Reduction Technology® treatment does not affect acute lung injury in a two-event in vivo model caused by stored blood components

    PubMed Central

    Silliman, C. C.; Khan, S. Y.; Ball, J. Bradley; Kelher, M. R.; Marschner, S.

    2011-01-01

    Introduction Mirasol Pathogen Reduction Technology® (PRT) treatment uses riboflavin and UV light to inactivate pathogens in blood components. Neutrophil [polymorphonuclear cells (PMN)] priming activity accumulates during routine storage of cellular blood components, and this activity has been implicated in transfusion-related acute lung injury (TRALI). We hypothesize that PRT-treatment of blood components affects the priming activity generated during storage of packed RBCs (PRBCs) or platelet concentrates (PCs), which can elicit ALI in vivo. Methods Plasma, PRBCs and PCs were isolated from healthy donor’s whole blood or by apheresis. Half of a collected unit was treated with PRT treatment and the remainder was left as an unmodified control. Supernatant was collected during storage of PCs and PRBCs and assayed for PMN priming activity and used as the second event in a two-event in vivo model of TRALI. Results PRT treatment did not induce priming activity in plasma or affect the priming activity generated during storage of PCs or PRBCs as compared with the unmodified controls. The supernatants from stored, but not fresh, PCs and PRBCs did cause ALI as the second event in a two-event animal model of TRALI, which was unaffected by PRT treatment. We conclude that the PRT® treatment does not induce priming activity in plasma nor does it affect the priming activity generated during storage of PCs or PRBCs or their ability to cause ALI as the second event in a two-event in vivo model of TRALI. Moreover, the amount of priming activity in TRIMA®-isolated PCs was significantly less than SPECTRA®-isolated PCs. PMID:19951305

  12. A retrospective study of factors which determine a negative blood culture in Cambodian children diagnosed with enteric fever

    PubMed Central

    Bousfield, Rachel; Thyl, Miliya; Samol, Orng; Rithea, Loet; Sona, Soeng; Chhat, Hor Put; Poda, Sar; Moore, Cartin E.; Chheng, Kheng; Kumar, Varun; Day, Nicholas P. J.; Parry, Christopher M.

    2016-01-01

    Background: Blood cultures are used to confirm a diagnosis of enteric fever but reported sensitivities can be as low as 40%. Aims: To determine the factors associated with a negative blood culture in Cambodian children with suspected enteric fever. Methods: In a retrospective study of hospitalised Cambodian children given a discharge diagnosis of enteric fever, the following factors associated with a negative blood culture were analysed: age, blood culture volume, prior antibiotic therapy, duration of illness and disease severity. Results: In 227 hospitalised Cambodian children with a discharge diagnosis of enteric fever, it was confirmed in 70% by a positive blood culture. There was no association between a negative blood culture and younger age, lower blood volumes for culture, prior antibiotic therapy, a late presentation or milder disease. Conclusions: Although blood culture sensitivity was higher than expected, alternative simple, rapid and sensitive tests are needed for diagnosing enteric fever. PMID:25845519

  13. Evaluation of PNA FISH® Yeast Traffic Light in identification of Candida species from blood and non-blood culture specimens.

    PubMed

    Radic, Marina; Goic-Barisic, Ivana; Novak, Anita; Rubic, Zana; Tonkic, Marija

    2016-08-01

    PNA FISH(®) (peptide nucleic acid fluorescent in situ hybridization) Yeast Traffic Light (PNA FISH(®) YTL) assay is a commercially avaliable method for rapid identification of Candida spp. directly from positive blood cultures. This report provides a one-year experience in identification of yeasts from 25 specimens (15 positive blood cultures and 10 other clinically significant specimens) using PNA FISH(®) YTL and comparing it to VITEK 2 System. Overall, assay identification compatibility with VITEK 2 System was found among 21/25 (84%) isolates tested. Only 3/25 (12%) of the isolates were not identified, and one isolate was misidentified by the PNA FISH(®) YTL assay. Our results show that the assay is a reliable method in identification of Candida spp. not only from blood cultures, but even from other clinically significant specimens (urine cultures, catheter tip cultures, peritoneal fluid cultures) when compared to automated method like VITEK 2 System. This novel application of the PNA FISH(®) YTL assay could therefore contribute to cost savings and significant benefit to patients, as rapid information about isolated yeast species is provided. © The Author 2016. Published by Oxford University Press on behalf of The International Society for Human and Animal Mycology. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.

  14. Minimizing the Workup of Blood Culture Contaminants: Implementation and Evaluation of a Laboratory-Based Algorithm

    PubMed Central

    Richter, S. S.; Beekmann, S. E.; Croco, J. L.; Diekema, D. J.; Koontz, F. P.; Pfaller, M. A.; Doern, G. V.

    2002-01-01

    An algorithm was implemented in the clinical microbiology laboratory to assess the clinical significance of organisms that are often considered contaminants (coagulase-negative staphylococci, aerobic and anaerobic diphtheroids, Micrococcus spp., Bacillus spp., and viridans group streptococci) when isolated from blood cultures. From 25 August 1999 through 30 April 2000, 12,374 blood cultures were submitted to the University of Iowa Clinical Microbiology Laboratory. Potential contaminants were recovered from 495 of 1,040 positive blood cultures. If one or more additional blood cultures were obtained within ±48 h and all were negative, the isolate was considered a contaminant. Antimicrobial susceptibility testing (AST) of these probable contaminants was not performed unless requested. If no additional blood cultures were submitted or there were additional positive blood cultures (within ±48 h), a pathology resident gathered patient clinical information and made a judgment regarding the isolate's significance. To evaluate the accuracy of these algorithm-based assignments, a nurse epidemiologist in approximately 60% of the cases performed a retrospective chart review. Agreement between the findings of the retrospective chart review and the automatic classification of the isolates with additional negative blood cultures as probable contaminants occurred among 85.8% of 225 isolates. In response to physician requests, AST had been performed on 15 of the 32 isolates with additional negative cultures considered significant by retrospective chart review. Agreement of pathology resident assignment with the retrospective chart review occurred among 74.6% of 71 isolates. The laboratory-based algorithm provided an acceptably accurate means for assessing the clinical significance of potential contaminants recovered from blood cultures. PMID:12089259

  15. Reducing Blood Culture Contamination in the Emergency Department: An Interrupted Time Series Quality Improvement Study

    PubMed Central

    Self, Wesley H.; Speroff, Theodore; Grijalva, Carlos G.; McNaughton, Candace D.; Ashburn, Jacki; Liu, Dandan; Arbogast, Patrick G.; Russ, Stephan; Storrow, Alan B.; Talbot, Thomas R.

    2012-01-01

    Objectives Blood culture contamination is a common problem in the emergency department (ED) that leads to unnecessary patient morbidity and health care costs. The study objective was to develop and evaluate the effectiveness of a quality improvement (QI) intervention for reducing blood culture contamination in an ED. Methods The authors developed a QI intervention to reduce blood culture contamination in the ED and then evaluated its effectiveness in a prospective interrupted times series study. The QI intervention involved changing the technique of blood culture specimen collection from the traditional clean procedure, to a new sterile procedure, with standardized use of sterile gloves and a new materials kit containing a 2% chlorhexidine skin antisepsis device, a sterile fenestrated drape, a sterile needle, and a procedural checklist. The intervention was implemented in a university-affiliated ED and its effect on blood culture contamination evaluated by comparing the biweekly percentages of blood cultures contaminated during a 48-week baseline period (clean technique), and 48-week intervention period (sterile technique), using segmented regression analysis with adjustment for secular trends and first-order autocorrelation. The goal was to achieve and maintain a contamination rate below 3%. Results During the baseline period, 321 out of 7,389 (4.3%) cultures were contaminated, compared to 111 of 6,590 (1.7%) during the intervention period (p < 0.001). In the segmented regression model, the intervention was associated with an immediate 2.9% (95% CI = 2.2% to 3.2%) absolute reduction in contamination. The contamination rate was maintained below 3% during each biweekly interval throughout the intervention period. Conclusions A QI assessment of ED blood culture contamination led to development of a targeted intervention to convert the process of blood culture collection from a clean to a fully sterile procedure. Implementation of this intervention led to an immediate

  16. Reducing blood culture contamination in the emergency department: an interrupted time series quality improvement study.

    PubMed

    Self, Wesley H; Speroff, Theodore; Grijalva, Carlos G; McNaughton, Candace D; Ashburn, Jacki; Liu, Dandan; Arbogast, Patrick G; Russ, Stephan; Storrow, Alan B; Talbot, Thomas R

    2013-01-01

    Blood culture contamination is a common problem in the emergency department (ED) that leads to unnecessary patient morbidity and health care costs. The study objective was to develop and evaluate the effectiveness of a quality improvement (QI) intervention for reducing blood culture contamination in an ED. The authors developed a QI intervention to reduce blood culture contamination in the ED and then evaluated its effectiveness in a prospective interrupted times series study. The QI intervention involved changing the technique of blood culture specimen collection from the traditional clean procedure to a new sterile procedure, with standardized use of sterile gloves and a new materials kit containing a 2% chlorhexidine skin antisepsis device, a sterile fenestrated drape, a sterile needle, and a procedural checklist. The intervention was implemented in a university-affiliated ED and its effect on blood culture contamination evaluated by comparing the biweekly percentages of blood cultures contaminated during a 48-week baseline period (clean technique) and 48-week intervention period (sterile technique), using segmented regression analysis with adjustment for secular trends and first-order autocorrelation. The goal was to achieve and maintain a contamination rate below 3%. During the baseline period, 321 of 7,389 (4.3%) cultures were contaminated, compared to 111 of 6,590 (1.7%) during the intervention period (p < 0.001). In the segmented regression model, the intervention was associated with an immediate 2.9% (95% confidence interval [CI] = 2.2% to 3.2%) absolute reduction in contamination. The contamination rate was maintained below 3% during each biweekly interval throughout the intervention period. A QI assessment of ED blood culture contamination led to development of a targeted intervention to convert the process of blood culture collection from a clean to a fully sterile procedure. Implementation of this intervention led to an immediate and sustained

  17. Recent Progress in the Diagnosis of Pathogenic Candida Species in Blood Culture.

    PubMed

    Phoompoung, Pakpoom; Chayakulkeeree, Methee

    2016-06-01

    Candidemia has become an emerging invasive fungal disease. Prompt treatment with appropriate antifungal agent is crucial to reduce the mortality of candidemia. The conventional blood culture method, which is considered the gold standard for candidemia diagnosis, has a low sensitivity and is time-consuming to perform. Recently, several novel advanced diagnostic methods that have a higher sensitivity and a shorter turnaround time than the conventional blood culture method have been developed for the early detection of Candida in blood samples or in blood culture broth. Most of these newer methods were developed using various molecular techniques, such as matrix-assisted laser desorption/ionization time-of-flight mass spectrometry, peptide nucleic acid fluorescence in situ hybridization, and a number of DNA-based techniques including in-house and commercial polymerase chain reactions. In this article, we review and summarize the novel molecular methods that have been recently used for the detection and identification of Candida organisms in blood specimens.

  18. Blood Culture Testing via a Mobile App That Uses a Mobile Phone Camera: A Feasibility Study.

    PubMed

    Lee, Guna; Lee, Yura; Chong, Yong Pil; Jang, Seongsoo; Kim, Mi Na; Kim, Jeong Hoon; Kim, Woo Sung; Lee, Jae-Ho

    2016-10-26

    To evaluate patients with fever of unknown origin or those with suspected bacteremia, the precision of blood culture tests is critical. An inappropriate step in the test process or error in a parameter could lead to a false-positive result, which could then affect the direction of treatment in critical conditions. Mobile health apps can be used to resolve problems with blood culture tests, and such apps can hence ensure that point-of-care guidelines are followed and processes are monitored for blood culture tests. In this pilot project, we aimed to investigate the feasibility of using a mobile blood culture app to manage blood culture test quality. We implemented the app at a university hospital in South Korea to assess the potential for its utilization in a clinical environment by reviewing the usage data among a small group of users and by assessing their feedback and the data related to blood culture sampling. We used an iOS-based blood culture app that uses an embedded camera to scan the patient identification and sample number bar codes. A total of 4 medical interns working at 2 medical intensive care units (MICUs) participated in this project, which spanned 3 weeks. App usage and blood culture sampling parameters (including sampler, sampling site, sampling time, and sample volume) were analyzed. The compliance of sampling parameter entry was also measured. In addition, the participants' opinions regarding patient safety, timeliness, efficiency, and usability were recorded. In total, 356/644 (55.3%) of all blood culture samples obtained at the MICUs were examined using the app, including 254/356 (71.3%) with blood collection volumes of 5-7 mL and 256/356 (71.9%) with blood collection from the peripheral veins. The sampling volume differed among the participants. Sampling parameters were completely entered in 354/356 cases (99.4%). All the participants agreed that the app ensured good patient safety, disagreed on its timeliness, and did not believe that it was

  19. Blood Culture Testing via a Mobile App That Uses a Mobile Phone Camera: A Feasibility Study

    PubMed Central

    Chong, Yong Pil; Jang, Seongsoo; Kim, Mi Na; Kim, Jeong Hoon; Kim, Woo Sung

    2016-01-01

    Background To evaluate patients with fever of unknown origin or those with suspected bacteremia, the precision of blood culture tests is critical. An inappropriate step in the test process or error in a parameter could lead to a false-positive result, which could then affect the direction of treatment in critical conditions. Mobile health apps can be used to resolve problems with blood culture tests, and such apps can hence ensure that point-of-care guidelines are followed and processes are monitored for blood culture tests. Objective In this pilot project, we aimed to investigate the feasibility of using a mobile blood culture app to manage blood culture test quality. We implemented the app at a university hospital in South Korea to assess the potential for its utilization in a clinical environment by reviewing the usage data among a small group of users and by assessing their feedback and the data related to blood culture sampling. Methods We used an iOS-based blood culture app that uses an embedded camera to scan the patient identification and sample number bar codes. A total of 4 medical interns working at 2 medical intensive care units (MICUs) participated in this project, which spanned 3 weeks. App usage and blood culture sampling parameters (including sampler, sampling site, sampling time, and sample volume) were analyzed. The compliance of sampling parameter entry was also measured. In addition, the participants’ opinions regarding patient safety, timeliness, efficiency, and usability were recorded. Results In total, 356/644 (55.3%) of all blood culture samples obtained at the MICUs were examined using the app, including 254/356 (71.3%) with blood collection volumes of 5-7 mL and 256/356 (71.9%) with blood collection from the peripheral veins. The sampling volume differed among the participants. Sampling parameters were completely entered in 354/356 cases (99.4%). All the participants agreed that the app ensured good patient safety, disagreed on its

  20. Detection of OXA-370 directly from rectal swabs and blood culture vials using an immunochromatographic assay.

    PubMed

    Nodari, Carolina Silva; Gales, Ana Cristina; Barth, Afonso Luís; Magagnin, Cibele Massotti; Zavascki, Alexandre Prehn; Carvalhaes, Cecília Godoy

    2017-08-01

    We evaluated the performance of OXA-48 K-SeT assay for detecting OXA-370 directly from spiked rectal swabs and blood culture vials. The limit of detection of this test was 10(4)UFC/mL for rectal swabs. Detection of the OXA-370-producing isolates was successfully achieved directly from positive blood culture vials independent of growing conditions. Copyright © 2017 Elsevier B.V. All rights reserved.

  1. Rapid Identification of Pathogens from Positive Blood Cultures by Multiplex PCR using the FilmArray System

    PubMed Central

    Blaschke, Anne J.; Heyrend, Caroline; Byington, Carrie L.; Fisher, Mark A.; Barker, Elizabeth; Garrone, Nicholas F.; Thatcher, Stephanie A.; Pavia, Andrew T.; Barney, Trenda; Alger, Garrison D.; Daly, Judy A.; Ririe, Kirk M.; Ota, Irene; Poritz, Mark A.

    2012-01-01

    Sepsis is a leading cause of death. Rapid and accurate identification of pathogens and antimicrobial resistance directly from blood culture could improve patient outcomes. The FilmArray® (FA; Idaho Technology, Inc., Salt Lake City, UT) Blood Culture (BC) panel can identify > 25 pathogens and 4 antibiotic resistance genes from positive blood cultures in 1 hour. We compared a development version of the panel to conventional culture and susceptibility testing on 102 archived blood cultures from adults and children with bacteremia. Of 109 pathogens identified by culture, 95% were identified by FA. Among 111 prospectively collected blood cultures, the FA identified 84 of 92 pathogens (91%) covered by the panel. Among 25 Staphylococcus aureus and 21 Enterococcus species detected, FA identified all culture-proven MRSA and VRE. The FA BC panel is an accurate method for the rapid identification of pathogens and resistance genes from blood culture. PMID:22999332

  2. A Retinol Isotope Dilution Equation Predicts Both Group and Individual Total Body Vitamin A Stores in Adults Based on Data from an Early Postdosing Blood Sample.

    PubMed

    Green, Michael H; Ford, Jennifer Lynn; Green, Joanne Balmer; Berry, Philip; Boddy, Alan V; Oxley, Anthony; Lietz, Georg

    2016-10-01

    Retinol isotope dilution (RID) is used to determine vitamin A total body stores (TBS) after an oral dose of a vitamin A stable isotope. The generally accepted prediction equation proposed by Olson's group in 1989 (Furr et al. Am J Clin Nutr 1989;49:713-6) includes factors related to dose absorption and retention, isotope equilibration in plasma compared with stores, catabolism during the mixing period, and the optimal time for measuring plasma isotope enrichment. The objectives were 1) to develop a modified RID equation and identify an earlier sampling time for predicting TBS and 2) to improve prediction in individuals as well as groups. To develop a modified RID equation, we used results of model-based compartmental analysis [the Simulation, Analysis and Modeling software (WinSAAM version 3.0.8; http://www.WinSAAM.org)] of plasma [(13)C10]retinol kinetic data from 32 previously studied, healthy young adults of European ancestry who had moderate vitamin A intakes and who ingested 2.95 μmol [(13)C10]retinyl acetate. We examined the time dependence of factors in the prediction equation related to absorption/retention (Fa) and isotope equilibration (S) and determined that 4 or 5 d postdosing was the optimal sampling time. TBS calculated by the equation TBS = Fa x S x (1/SAp), where SAp is plasma retinol specific activity (fraction of dose/μmol), were highly correlated with model-predicted TBS (r = 0.95 and 0.96 for 4 and 5 d, respectively; P < 0.001); predictions for individuals were also highly correlated (Rs = 0.94 and 0.94; P < 0.001). The equation TBS ≈ 0.5 × (1/SAp) accurately predicted vitamin A TBS in this group of 32 healthy young adults and its individual members with the use of data from 1 blood sample taken 4 d after isotope administration.

  3. A novel procedure to improve functional preservation of hematopoietic stem and progenitor cells in cord blood stored at +4°c before cryopreservation.

    PubMed

    Chevaleyre, Jean; Rodriguez, Laura; Duchez, Pascale; Plainfossé, Marie; Dazey, Bernard; Lapostolle, Véronique; Vlaski, Marija; Brunet de la Grange, Philippe; Delorme, Bruno; Ivanovic, Zoran

    2014-08-01

    During storage and transportation of collected cord blood units (CBUs) to the bank prior to their processing and cryopreservation, it is imperative to preserve the functional capacities of a relatively small amount of cells of interest (stem and progenitor cells) which are critical for graft potency. To improve CBU storage efficiency, we conceived an approach based on the following two principles: (1) to provide a better nutritive and biochemical environment to stem and progenitor cells in CB and (2) to prevent the hyperoxygenation of these cells transferred from a low- (1.1%-4% O2 in the CB) to a high-oxygen (20%-21% O2 in atmosphere) concentration. Our hypothesis is confirmed by the functional assessment of stem cell (hematopoietic reconstitution capacity in immunodeficient mice-scid repopulating cell assay) and committed progenitor activities (capacity of in vitro colony formation and of ex vivo expansion) after the storage period with our medium (HP02) in gas-impermeable bags. This storage procedure maintains the full functional capacity of a CBU graft for 3 days with respect to day 0. Further, using this procedure, a graft stored 3 days at +4°C exhibits better functional capacities than one currently used in routine storage (CBUs stored at +4°C for 1 day in gas-permeable bags and without medium). We provided the proof of principle of our approach, developed a clinical-scale kit and performed a preclinical assay demonstrating the feasibility and efficiency of our CBU preservation protocol through all steps of preparation (volume reduction, freezing, and thawing).

  4. The effect of using blood culture bottle of bronchoalveolar larvage fluid in pneumonia.

    PubMed

    Heo, Eun Young; Shin, Sue; Chung, Hee Soon; Jeong, Yun-Jeong; Oh, So Hee; Kim, Deog Kyeom

    2016-06-06

    Pneumonia is a primary cause of morbidity and mortality in infectious disease, and increasing antimicrobial resistance has raised concerns of treatment failure. Therefore, we evaluated the value of a blood culture bottle for bronchoalveolar lavage (BAL) samples on pathogen identification and on treatment modification in patients with pneumonia. We conducted a prospective study and enrolled 39 patients who were hospitalized for pneumonia. Enrolled patients underwent BAL; a 10-ml aliquot was transferred to a sterile container for standard quantitative culture, and a 5 ml aliquot was transferred to both an aerobic and an anaerobic blood culture bottle. Microbes were detected in all 39 (100 %) specimens and possible pathogens were identified in 34 patients (84.6 %) from BAL blood culture bottles. In contrast, microbes were detected in 10 patients (25.6 %) and possible pathogens were isolated in 8 patients (20.5 %) in BAL fluid using conventional culture methods. Finally, 8 of 39 (20.5 %) patients changed antibiotics according to the BAL blood culture results and pneumonia improved in 6 of these patients. Using blood culture bottles for BAL sampling in patients with pneumonia is a sensitive method to detect pathogens in order to identify an adequate antibiotic treatment regimen.

  5. MALDI-TOF mass spectrometry for early identification of bacteria grown in blood culture bottles.

    PubMed

    Zabbe, Jean-Benoît; Zanardo, Laura; Mégraud, Francis; Bessède, Emilie

    2015-08-01

    This note reports an interesting way to rapidly identify bacteria grown from blood culture bottles. Chocolate agar plates were inoculated with 1 drop of the positive blood bottle medium. After a 3-hour incubation, the growth veil was submitted to MALDI-TOF mass spectrometry: 77% of the bacteria present have been correctly identified.

  6. Effect of the addition of the antioxidant taurine on the complete blood count of whole blood stored at room temperature and at 4ºC for up to 7 days.

    PubMed

    Sirdah, Mahmoud Mohammed; Abushahla, Abdelnasser Kassem; Al-Sarraj, Heba Abd Allah

    2013-01-01

    The complete blood count is one of the most common routine tests. This study aimed to evaluate possible effects of the antioxidant taurine on the complete blood count of whole blood stored at room temperature and at 4ºC over seven days. Venous blood samples of 25 healthy males were distributed into two sets of tubes with each set of four tubes containing 50 µL of solutions with zero, 2.5 g/L, 5 g/L, 10 g/L taurine. The tubes were kept at room temperature or at 4ºC. Complete blood counts were performed on seven successive days. The mean percentage changes [Δ = (mean value - mean baseline value) / mean baseline value x 100] were calculated and compared. Complete blood count parameters exhibited different patterns of behavior which were affected by the storage temperature, time and taurine concentration. Taurine at room temperature significantly enhanced the stability of: the platelet count over seven days (Δ7 at 2.5, 5 and 10 g/L taurine were 5.45, 6.11, and 5.80 x 10(9) cells/L, respectively); the red blood cell count over five days (Δ5 at 2.5, 5 and 10 g/L taurine were 1.59, 2.79, and 1.98 x 10(12) cells/L, respectively); mean corpuscular hemoglobin over five days (Δ5 at 2.5, 5 and 10 g/L taurine were -0.91,-1.52 and -0.84 fl respectively); and red cell distribution width over two days (Δ2 at 2.5, 5 and 10 g/L taurine were 0.90%, 1.30% and -0.1%, respectively). No additional stabilizing effects of taurine were reported for the mean corpuscular volume, mean corpuscular hemoglobin, mean corpuscular hemoglobin concentration, hematocrit and hemoglobin, while it negatively affected the white blood cell stability. Complete blood count parameters exhibited variable stability patterns in respect to temperature, time and taurine concentration.

  7. Linking Positive Affect to Blood Lipids: A Cultural Perspective.

    PubMed

    Yoo, Jiah; Miyamoto, Yuri; Rigotti, Attilio; Ryff, Carol D

    2017-10-01

    Higher levels of positive affect have been associated with better physical health. While positive affect is seen as highly desirable among Westerners, East Asians tend to deemphasize positive affect. Using large probability samples of Japanese and U.S. adult populations, the present study examined the relations of positive affect with serum lipid profiles, known to be strongly predictive of risk for cardiovascular disease, and tested whether their associations depend on cultural contexts. As predicted, positive affect was associated with healthier lipid profiles for Americans but not for Japanese. Further analyses showed that this cultural moderation was mediated by body mass index. This study highlights the role of culture in the link between positive emotions and key biological risk factors of cardiovascular disease.

  8. Hemoglobin Function in Stored Blood.

    DTIC Science & Technology

    1974-08-01

    and inosine with or without methylene blue , showed that the pH 6.4 to 7.2 preservatives afforded the best DPG maintenance. 2. Experiments with CPD...adenine-inosine with and without uiethylene blue indicate that the methylene blue effect is dependent on the presence of inosine for maintenance of 2,3...this country and in transfusion practice in several countries in Europe for a number of years. It in believed that the work involving methylene - blue represents

  9. Hemoglobin Function in Stored Blood.

    DTIC Science & Technology

    1979-01-01

    glucose concentrations, adenine, inosine, methylene blue , phosphate, pyruvate, dihydroxyacetone, ascorbic acid, fructose, mannose, galactose, and ribose. Also, D-ascorbate, dehydroascorbate and SH inhibitors. (Author)

  10. Effects of carbon sources on the enrichment of halophilic polyhydroxyalkanoate-storing mixed microbial culture in an aerobic dynamic feeding process

    NASA Astrophysics Data System (ADS)

    Cui, You-Wei; Zhang, Hong-Yu; Lu, Peng-Fei; Peng, Yong-Zhen

    2016-08-01

    Microbial polyhydroxyalkanoate (PHA) production serves as a substitute for petroleum-based plastics. Enriching mixed microbial cultures (MMCs) with the capacity to store PHA is a key precursor for low-cost PHA production. This study investigated the impact of carbon types on enrichment outcomes. Three MMCs were separately fed by acetate sodium, glucose, and starch as an enriching carbon source, and were exposed to long-term aerobic dynamic feeding (ADF) periods. The PHA production capacity, kinetics and stoichiometry of the enrichments, the PHA composition, and the microbial diversity and community composition were explored to determine carbon and enrichment correlations. After 350-cycle enriching periods under feast-famine (F-F) regimes, the MMCs enriched by acetate sodium and glucose contained a maximum PHA content of 64.7% and 60.5% cell dry weight (CDW). The starch-enriched MMC only had 27.3% CDW of PHA. High-throughput sequencing revealed that non-PHA bacteria survived alongside PHA storing bacteria, even under severe F-F selective pressure. Genus of Pseudomonas and Stappia were the possible PHA accumulating bacteria in acetate-enriched MMC. Genus of Oceanicella, Piscicoccus and Vibrio were found as PHA accumulating bacteria in glucose-enriched MMC. Vibrio genus was the only PHA accumulating bacteria in starch-enriched MMC. The community diversity and composition were regulated by the substrate types.

  11. Effects of carbon sources on the enrichment of halophilic polyhydroxyalkanoate-storing mixed microbial culture in an aerobic dynamic feeding process

    PubMed Central

    Cui, You-Wei; Zhang, Hong-Yu; Lu, Peng-Fei; Peng, Yong-Zhen

    2016-01-01

    Microbial polyhydroxyalkanoate (PHA) production serves as a substitute for petroleum-based plastics. Enriching mixed microbial cultures (MMCs) with the capacity to store PHA is a key precursor for low-cost PHA production. This study investigated the impact of carbon types on enrichment outcomes. Three MMCs were separately fed by acetate sodium, glucose, and starch as an enriching carbon source, and were exposed to long-term aerobic dynamic feeding (ADF) periods. The PHA production capacity, kinetics and stoichiometry of the enrichments, the PHA composition, and the microbial diversity and community composition were explored to determine carbon and enrichment correlations. After 350-cycle enriching periods under feast-famine (F-F) regimes, the MMCs enriched by acetate sodium and glucose contained a maximum PHA content of 64.7% and 60.5% cell dry weight (CDW). The starch-enriched MMC only had 27.3% CDW of PHA. High-throughput sequencing revealed that non-PHA bacteria survived alongside PHA storing bacteria, even under severe F-F selective pressure. Genus of Pseudomonas and Stappia were the possible PHA accumulating bacteria in acetate-enriched MMC. Genus of Oceanicella, Piscicoccus and Vibrio were found as PHA accumulating bacteria in glucose-enriched MMC. Vibrio genus was the only PHA accumulating bacteria in starch-enriched MMC. The community diversity and composition were regulated by the substrate types. PMID:27485896

  12. Educational intervention as an effective step for reducing blood culture contamination: a prospective cohort study.

    PubMed

    Park, W B; Myung, S J; Oh, M-d; Lee, J; Kim, N-J; Kim, E-C; Park, J S

    2015-10-01

    Contaminated blood cultures lead to diagnostic challenges and place a burden on healthcare services. To determine the impact of introducing a clinical skills test (CST) as part of the medical licensing examination and an institutional education programme on the contamination rates of blood cultures. A prospective cohort study was conducted from 2009 through 2013 in all wards of a tertiary-care teaching hospital. We evaluated the effects of the CST, which was added to the National Medical Licensing Examination in Korea (KMLE) in 2010 and our institutional education programme, which began in 2013. The medical interns in charge of collection of blood for culture were divided in three groups with presence or absence of CST and the institutional education programme. The primary outcome was the percentage of blood cultures contaminated in each group, which were compared using the Poisson regression model. Participants' self-rated scores for the blood draw procedure were also analysed. Although introduction of the CST in the KMLE failed to reduce blood culture contamination rate (1.36% vs 1.35%; P = 0.734), the institutional education programme significantly reduced the contamination rate (1.35% vs 1.00%; P < 0.0001). Most participants answered that they always followed each step correctly except for waiting the recommended contact time after applying the antiseptic. The educational intervention, not the introduction of CST in the KMLE, was effective in reducing overall contamination rates. Copyright © 2015 The Healthcare Infection Society. Published by Elsevier Ltd. All rights reserved.

  13. Reduced length of hospital stay through a point of care placed automated blood culture instrument.

    PubMed

    Bruins, M J; Egbers, M J; Israel, T M; Diepeveen, S H A; Wolfhagen, M J H M

    2017-04-01

    Early appropriate antimicrobial treatment of patients with sepsis has a large impact on clinical outcome. To enable prompt and efficient processing of blood cultures, the inoculated vials should be placed into an automated continuously monitoring blood culture system immediately after sampling. We placed an extra BACTEC FX instrument at the emergency department of our hospital and validated the twice-daily re-entering of ongoing vials from this instrument into the BACTEC FX at the laboratory. We subsequently assessed the benefits of shortening the transport time between sampling and monitored incubation of blood culture vials by comparing the turnaround times of positive blood cultures from emergency department patients with a historical control group. Re-entering ongoing vials within 2 h raised no technical problems with the BACTEC FX and did not increase the risk of false-negative culture results. The decreased transport time resulted in significantly earlier available Gram stain results for a large proportion of patients in the intervention group and a significant shortening of the median total turnaround time to less than 48 h. The median length of hospital stay shortened by 1 day. Immediate entering of blood culture vials into a point of care placed BACTEC FX instrument and subsequent efficient processing enables earlier decision-making regarding antimicrobial treatment, preventing the development of antimicrobial resistance and reducing healthcare costs.

  14. Effect of Cultural Environment on the Blood Group Activity of Microorganisms1

    PubMed Central

    Moody, Marcia R.; Young, Viola M.; Faber, John E.

    1969-01-01

    Blood group activity was proven to be a property of the bacterium per se which possesses it, although such activity was influenced by the cultural environment of the organism. High concentrations of peptones having blood group activity were able to transfer this activity to inactive organisms; however, the conferred activity was proportional to the concentration. As a result of the low concentrations, the blood group activity of peptones was eliminated upon incorporation in culture media, and the activity of the peptones had no effect on the blood group activity levels of microorganisms when grown in these media. Conversely, the vitamin content of culture media did affect blood group active organisms. Multiple vitamins in media decreased the activity levels by blocking the reactive sites of the active organisms on which activity detection depended. Blood group activity levels were highest in media of minimal or no vitamin content. Therefore, it can be concluded that a choice of cultural medium becomes an important factor in the quantitation of bacterial blood group activity. PMID:4896884

  15. Reconstitution activity of hypoxic cultured human cord blood CD34-positive cells in NOG mice

    SciTech Connect

    Shima, Haruko; Takubo, Keiyo; Iwasaki, Hiroko; Yoshihara, Hiroki; Gomei, Yumiko; Hosokawa, Kentaro; Arai, Fumio; Takahashi, Takao; Suda, Toshio

    2009-01-16

    Hematopoietic stem cells (HSCs) reside in hypoxic areas of the bone marrow. However, the role of hypoxia in the maintenance of HSCs has not been fully characterized. We performed xenotransplantation of human cord blood cells cultured in hypoxic or normoxic conditions into adult NOD/SCID/IL-2R{gamma}{sup null} (NOG) mice. Hypoxic culture (1% O{sub 2}) for 6 days efficiently supported the maintenance of HSCs, although cell proliferation was suppressed compared to the normoxic culture. In contrast, hypoxia did not affect in vitro colony-forming ability. Upregulation of a cell cycle inhibitor, p21, was observed in hypoxic culture. Immunohistochemical analysis of recipient bone marrow revealed that engrafted CD34{sup +}CD38{sup -} cord blood HSCs were hypoxic. Taken together, these results demonstrate the significance of hypoxia in the maintenance of quiescent human cord blood HSCs.

  16. Evaluation of three rapid diagnostic methods for direct identification of microorganisms in positive blood cultures.

    PubMed

    Martinez, Raquel M; Bauerle, Elizabeth R; Fang, Ferric C; Butler-Wu, Susan M

    2014-07-01

    The identification of organisms from positive blood cultures generally takes several days. However, recently developed rapid diagnostic methods offer the potential for organism identification within only a few hours of blood culture positivity. In this study, we evaluated the performance of three commercial methods to rapidly identify organisms directly from positive blood cultures: QuickFISH (AdvanDx, Wolburn, MA), Verigene Gram-Positive Blood Culture (BC-GP; Nanosphere, Northbrook, IL), and matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) with Sepsityper processing (Bruker Daltonics, Billerica, MA). A total of 159 blood cultures (VersaTREK Trek Diagnostic Systems, Cleveland, OH) positive for Gram-positive and Gram-negative bacteria as well as yeast were analyzed with QuickFISH and MALDI-TOF MS. In all, 102 blood cultures were analyzed using the BC-GP assay. For monomicrobial cultures, we observed 98.0% concordance with routine methods for both QuickFISH (143/146) and the BC-GP assay (93/95). MALDI-TOF MS demonstrated 80.1% (117/146) and 87.7% (128/146) concordance with routine methods to the genus and species levels, respectively. None of the methods tested were capable of consistently identifying polymicrobial cultures in their entirety or reliably differentiating Streptococcus pneumoniae from viridans streptococci. Nevertheless, the methods evaluated in this study are convenient and accurate for the most commonly encountered pathogens and have the potential to dramatically reduce turnaround time for the provision of results to the treating physician.

  17. Blood culture series benefit may be limited to selected clinical conditions: time to reassess.

    PubMed

    Khatib, R; Simeunovic, G; Sharma, M; Fakih, M G; Johnson, L B; Briski, L; Lebar, W

    2015-04-01

    Blood cultures are often submitted as series (two to three sets per 24 hours) to maximize sample recovery. We assessed the actual benefit of additional sets. Blood cultures submitted from adults (≥ 18 years old) over 1 year (1 February 2012 to 31 January 2013) were examined. The medical records of patients with positive cultures were reviewed. Cultures with commensal organisms were considered contamination in the absence of a source and clinical findings. The impact of additional sets on antibiotic therapy was estimated. We evaluated 15,394 blood cultures. They were submitted as two to five sets per 24 hours in 12,236 (79.5%) instances. Pathogens were detected in 1227 sets, representing 741 bacteremias, of which 618 (83.4%) were detected in the first set and 123 (16.6%) in the additional sets. Pathogens missed in the first set were recovered from patients receiving antibiotics (n = 72; 58.5%) and after undergoing a procedure (n = 54; 43.9%). The additional sets' results could have influenced antibiotic therapy in 76/6235 (1.2%) instances, including 40 (0.6%) antibiotic switches and 36 (0.6%) possible extensions of therapy. The potential impact of the detection of missed pathogens on antibiotic therapy was not apparent in patients who had an endovascular infection (26/27, 96.3%) and those who lacked an obvious source of pathogens (10/10, 100%). These findings suggest that one blood culture is probably adequate in patients with an obvious source of pathogens. Blood culture series are beneficial in patients without an obvious source of pathogens and in those with endovascular infections. It is time to reassess the benefit of blood culture series, perhaps limiting them to selected conditions.

  18. Association of a Clinical Practice Guideline With Blood Culture Use in Critically Ill Children.

    PubMed

    Woods-Hill, Charlotte Z; Fackler, James; Nelson McMillan, Kristen; Ascenzi, Judith; Martinez, Diego A; Toerper, Matthew F; Voskertchian, Annie; Colantuoni, Elizabeth; Klaus, Sybil Ann; Levin, Scott; Milstone, Aaron M

    2017-02-01

    Sepsis and septic shock are common and, at times, fatal in pediatrics. Blood cultures are often obtained when clinicians suspect sepsis, yet are low-yield with a false-positive rate up to 50%. To determine whether a novel, 2-part, clinical practice guideline could decrease the rates of total blood cultures and cultures collected from central venous catheters in critically ill children and to examine the effect of the guideline on patient outcomes. A retrospective cohort study was performed to determine the effect of a new clinical practice guideline on blood culture practices in a 36-bed, combined medical/surgical pediatric intensive care unit of an urban, academic, tertiary care center from April 1, 2013, to March 31, 2015. All patients admitted to the pediatric intensive care unit with length of stay of 4 hours or more were evaluated (4560 patient visits: 2204 preintervention, 2356 postintervention visits). Two documents were developed: (1) fever/sepsis screening checklist and (2) blood culture decision algorithm. Clinicians consulted these documents when considering ordering blood cultures and for guidance about the culture source. Primary outcome was the total number of blood cultures collected per 100 patient-days. Of the 2204 children evaluated before the intervention, 1215 were male (55.1%); median (interquartile range) age was 5 (1-13) years. Postintervention analysis included 2356 children; 1262 were male (53.6%) and median (interquartile range) age was 6 (1-13) years. A total of 1807 blood cultures were drawn before the intervention during 11 196 patient-days; 984 cultures were drawn after the intervention during 11 204 patient-days (incidence rate, 16.1 vs 8.8 cultures per 100 patient-days). There was a 46.0% reduction after the intervention in the blood culture collection rate (incidence rate ratio, 0.54; 95% CI, 0.50-0.59). After the intervention, there was an immediate 25.0% reduction in the rate of cultures per 100 patient-days (95% CI, 4

  19. Storing Hydrogen

    SciTech Connect

    Kim, Hyun Jeong; Karkamkar, Abhijeet J.; Autrey, Thomas; Chupas, Peter; Proffen, Thomas E.

    2010-05-31

    Researchers have been studying mesoporous materials for almost two decades with a view to using them as hosts for small molecules and scaffolds for molding organic compounds into new hybrid materials and nanoparticles. Their use as potential storage systems for large quantities of hydrogen has also been mooted. Such systems that might hold large quantities of hydrogen safely and in a very compact volume would have enormous potential for powering fuel cell vehicles, for instance. A sponge-like form of silicon dioxide, the stuff of sand particles and computer chips, can soak up and store other compounds including hydrogen. Studies carried out at the XOR/BESSRC 11-ID-B beamline at the APS have revealed that the nanoscopic properties of the hydrogenrich compound ammonia borane help it store hydrogen more efficiently than usual. The material may have potential for addressing the storage issues associated with a future hydrogen economy. Pacific Northwest National Laboratory is operated by Battelle for the US Department of Energy.

  20. Analysis of Anaerobic Blood Cultures in Burned Patients

    DTIC Science & Technology

    2007-01-01

    34 3 (9.0) Escherichia coli 14 (4.1) 17 (2.8) – – Enterobacter cloacae 13 (3.8) 24 (3.9) 18 2 (11.1) Enterobacter aerogenes 13 (3.8) 18 (2.9...cultures: patient characteristics and potential risk factors. Clin Chem Lab Med 2003;41(3):293–7. [10] James PA, Al-Shafi KM. Clinical value of anaerobic

  1. Resistance profiles of coagulase-negative staphylococci contaminating blood cultures predict pathogen resistance and patient mortality.

    PubMed

    Obolski, Uri; Alon, Danny; Hadany, Lilach; Stein, Gideon Y

    2014-09-01

    Blood culture isolates are the cornerstone of adequate antibiotic treatment. However, many blood cultures are contaminated with bacteria residing on the skin, the most common contaminants being coagulase-negative staphylococci (CoNS). Such contaminated cultures are mostly disregarded. In this retrospective study, we show that contaminated cultures contain diagnostic information. We tested the association between resistance profiles of CoNS contaminants and those of the actual infecting bacteria isolated subsequently from the same patient, as well as their association with short-term mortality. We identified all patients in Rabin Medical Center, Israel, with positive blood cultures during 2009-12. Data included patient demographics, hospitalization records, comorbidities, blood culture results and date of death. Our cohort consists of 2518 patients with 5290 blood cultures, where 1124 patients had 1664 blood cultures with CoNS contaminants. High overall CoNS resistance predicted high overall resistance of the subsequent bacterial isolates (P<0.004 and P<0.0006, for Gram-positive and -negative bacteria, respectively). Moreover, the resistance of CoNS contaminants to a specific antibiotic predicted the resistance of the subsequent bacterial isolates to that antibiotic (OR=5.55, 95% CI=3.54-8.66, P<10(-15) and OR=2.47, 95% CI=1.61-3.78, P<3 ×10(-5), for Gram-positive and -negative bacteria, respectively). Finally, highly resistant CoNS isolates were associated with higher short-term mortality (hazard ratio=1.71, 95% CI=1.4-2.11, P<10(-6)). Resistance patterns of CoNS contaminants predict specific and overall resistance of subsequent blood culture isolates and short-term mortality. These results may help predict patient mortality and correct empirical antibiotic therapy if blood cultures yield contaminant bacteria and imply that skin commensals may serve as an additional, non-invasive, diagnostic tool. © The Author 2014. Published by Oxford University Press on

  2. Fluorescent In Situ Hybridization Allows Rapid Identification of Microorganisms in Blood Cultures

    PubMed Central

    Kempf, Volkhard A. J.; Trebesius, Karlheinz; Autenrieth, Ingo B.

    2000-01-01

    Using fluorescent in situ hybridization (FISH) with rRNA-targeted fluorescently labelled oligonucleotide probes, pathogens were rapidly detected and identified in positive blood culture bottles without cultivation and biotyping. In this study, 115 blood cultures with a positive growth index as determined by a continuous-reading automated blood culture system were examined by both conventional laboratory methods and FISH. For this purpose, oligonucleotide probes that allowed identification of approximately 95% of those pathogens typically associated with bacteremia were produced. The sensitivity and specificity of these probes were 100%. From all 115 blood cultures, microorganisms were grown after 1 day and identification to the family, genus, or species level was achieved after 1 to 3 days while 111 samples (96.5%) were similarly identified by FISH within 2.5 h. Staphylococci were identified in 62 of 62 samples, streptococci and enterococci were identified in 19 of 20 samples, gram-negative rods were identified in 28 of 30 samples, and fungi were identified in two of two samples. Thus, FISH is an appropriate method for identification of pathogens grown in blood cultures from septicemic patients. PMID:10655393

  3. [Species and drug resistance of pathogens in blood cultures from the pediatric hematology ward].

    PubMed

    Kuang, Ling-Han; Jiang, Yong-Mei; Hu, Zheng-Qiang; Mu, Li-Yuan; Su, Min; Zhou, Wei

    2013-04-01

    To investigate the species and percentage changes of pathogens in blood cultures from the pediatric hematology ward, and to analyze the drug resistance of main pathogens and the risk factors for positive blood culture (sepsis). A retrospective analysis was performed to analyze the species and drug sensitivity of the pathogens isolated from 2358 blood cultures from the pediatric hematology ward of the West China Second University Hospital between 2008 and 2011, as well as the related clinical data. A total of 110 strains of pathogens were isolated, with Escherichia coli (16 strains), Pseudomonas aeruginosa (12 strains) and Staphylococcus epidermidis (8 strains) being the most common ones. From 2008 to 2011, the percentage of Gram-positive bacteria decreased, while the percentage of Gram-negative bacteria increased. The detection rates of methicillin-resistant coagulase-negative Staphylococci and methicillin-resistant Staphylococcus aureus were 69% and 43% respectively, but both were sensitive to vancomycin. The detection rates of extended-spectrum beta-lactamase (ESBL)-producing Escherichia coli and ESBL-producing Klebsiella pneumoniae were 69% and 62% respectively, but both were sensitive to imipenem and meropenem. Malignant tumor was a risk factor for positive blood culture (OR=3.564, P<0.05). A wide range of pathogens are responsible for bloodstream infection in the pediatric hematology ward and the percentages of bacteria are changing; these pathogens have a high drug resistance rate. Malignant tumor is a risk factor for positive blood culture in the pediatric hematology ward.

  4. Risk Factors for Neonatal Sepsis and Method for Reduction of Blood Culture Contamination.

    PubMed

    Krajčinović, S S; Doronjski, A; Barišić, N; Stojanović, V

    2015-03-01

    False-positive blood cultures findings may lead to a falsely increased morbidity and increased hospital costs. The survey was conducted as retrospective - prospective study and included 239 preterm infants (born before 37 weeks of gestation) who were treated in Neonatal Intensive Care Unit (NICU) in Institute for Child and Youth Health Care of Vojvodina during one year (January 1st, 2012 to December 31st, 2012). The retrospective part of the study focused on examination of incidence of neonatal sepsis and determination of risk factors. In the prospective part of the study infants were sub-divided into two groups: Group 1- infants hospitalized in NICU during the first 6 months of the study; blood cultures were taken by the "clean technique" and checklists for this procedure were not taken. Group 2- neonates hospitalized in NICU during last 6 months of the study; blood cultures were taken by "sterile technique" and checklists for this procedure were taken. The main risk factors for sepsis were prelabor rupture of membranes, low gestational age, low birth weight, mechanical ventilation, umbilical venous catheter placement, and abdominal drainage. Staphylococcus aureus and coagulase negative Staphylococcus were the most frequently isolated microorganisms in false-positive blood samples. Education of employees, use of checklists and sterile sets for blood sampling, permanent control of false positive blood cultures, as well as regular and routine monthly reports are crucial for successful reduction of contamination rates.

  5. Direct isolation of Candida spp. from blood cultures on the chromogenic medium CHROMagar Candida.

    PubMed

    Horvath, Lynn L; Hospenthal, Duane R; Murray, Clinton K; Dooley, David P

    2003-06-01

    CHROMagar Candida is a selective and differential chromogenic medium that has been shown to be useful for identification of Candida albicans, Candida krusei, Candida tropicalis, and perhaps Candida glabrata. Colony morphology and color have been well defined when CHROMagar Candida has been used to isolate yeast directly from clinical specimens, including stool, urine, respiratory, vaginal, oropharyngeal, and esophageal sources. Direct isolation of yeast on CHROMagar Candida from blood cultures has not been evaluated. We evaluated whether the color and colony characteristics produced by Candida spp. on CHROMagar Candida were altered when yeasts were isolated directly from blood cultures. Fifty clinical isolates of Candida were inoculated into aerobic and anaerobic blood culture bottles and incubated at 35 degrees C in an automated blood culture system. When growth was detected, an aliquot was removed and plated onto CHROMagar Candida. As a control, CHROMagar Candida plates were inoculated with the same isolate of yeast grown on Sabouraud dextrose agar simultaneously. No significant difference was detected in color or colony morphology between the blood and control isolates in any of the tested organisms. All C. albicans (n = 12), C. tropicalis (n = 12), C. glabrata (n = 9), and C. krusei (n = 5) isolates exhibited the expected species-specific colony characteristics and color, whether isolated directly from blood or from control cultures. CHROMagar Candida can be reliably used for direct isolation of yeast from blood cultures. Direct isolation could allow mycology laboratories to more rapidly identify Candida spp., enable clinicians to more quickly make antifungal agent selections, and potentially decrease patient morbidity and mortality.

  6. Direct Isolation of Candida spp. from Blood Cultures on the Chromogenic Medium CHROMagar Candida

    PubMed Central

    Horvath, Lynn L.; Hospenthal, Duane R.; Murray, Clinton K.; Dooley, David P.

    2003-01-01

    CHROMagar Candida is a selective and differential chromogenic medium that has been shown to be useful for identification of Candida albicans, Candida krusei, Candida tropicalis, and perhaps Candida glabrata. Colony morphology and color have been well defined when CHROMagar Candida has been used to isolate yeast directly from clinical specimens, including stool, urine, respiratory, vaginal, oropharyngeal, and esophageal sources. Direct isolation of yeast on CHROMagar Candida from blood cultures has not been evaluated. We evaluated whether the color and colony characteristics produced by Candida spp. on CHROMagar Candida were altered when yeasts were isolated directly from blood cultures. Fifty clinical isolates of Candida were inoculated into aerobic and anaerobic blood culture bottles and incubated at 35°C in an automated blood culture system. When growth was detected, an aliquot was removed and plated onto CHROMagar Candida. As a control, CHROMagar Candida plates were inoculated with the same isolate of yeast grown on Sabouraud dextrose agar simultaneously. No significant difference was detected in color or colony morphology between the blood and control isolates in any of the tested organisms. All C. albicans (n = 12), C. tropicalis (n = 12), C. glabrata (n = 9), and C. krusei (n = 5) isolates exhibited the expected species-specific colony characteristics and color, whether isolated directly from blood or from control cultures. CHROMagar Candida can be reliably used for direct isolation of yeast from blood cultures. Direct isolation could allow mycology laboratories to more rapidly identify Candida spp., enable clinicians to more quickly make antifungal agent selections, and potentially decrease patient morbidity and mortality. PMID:12791890

  7. Improved Amplification of Microbial DNA from Blood Cultures by Removal of the PCR Inhibitor Sodium Polyanetholesulfonate

    PubMed Central

    Fredricks, David N.; Relman, David A.

    1998-01-01

    Molecular methods are increasingly used to identify microbes in clinical samples. A common technical problem with PCR is failed amplification due to the presence of PCR inhibitors. Initial attempts at amplification of the bacterial 16S rRNA gene from inoculated blood culture media failed for this reason. The inhibitor persisted, despite numerous attempts to purify the DNA, and was identified as sodium polyanetholesulfonate (SPS), a common additive to blood culture media. Like DNA, SPS is a high-molecular-weight polyanion that is soluble in water but insoluble in alcohol. Accordingly, SPS tends to copurify with DNA. An extraction method was designed for purification of DNA from blood culture media and removal of SPS. Blood culture media containing human blood and spiked with Escherichia coli was subjected to an organic extraction procedure with benzyl alcohol, and removal of SPS was documented spectrophotometrically. Successful amplification of the extracted E. coli 16S rRNA gene was achieved by adding 5 μl of undiluted processed sample DNA to a 50-μl PCR mixture. When using other purification methods, the inhibitory effect of SPS could be overcome only by dilution of these samples. By our extraction technique, even uninoculated blood culture media were found to contain bacterial DNA when they were subjected to broad-range 16S rRNA gene consensus PCR. We conclude that the blood culture additive SPS is a potent inhibitor of PCR, is resistant to removal by traditional DNA purification methods, but can be removed by a benzyl alcohol extraction protocol that results in improved PCR performance. PMID:9738025

  8. Detection of Mycobacterium abscessus from blood cultures during treatment of interstitial pneumonia: a case study.

    PubMed

    Suzuki, Tomomi; Ito, Wataru; Takeda, Masahide; Kobayashi, Noriko; Uek, Shigeharu; Sato, Kazuhiro; Nakamura, Mio; Tomita, Noriko; Kayaba, Hiroyuki; Chihara, Junichi

    2011-09-01

    A 76-year-old male diagnosed with interstitial pneumonia in December 2002 was treated with a steroid in a nearby hospital. Exacerbation of infectious pneumonitis and interstitial pneumonia required complementary inpatient treatment in August 2007. Although polymerase chain reaction examination of expectorated sputa revealed the absence of Mycobacterium tuberculosis, M. avium, and M. intracellulare on admission, nontuberculous M. abscessus was detected in the routine blood cultures. Taken together with clinical findings, M. abscessus was most likely the primary causative organism. Diagnosis of mycobaterium-induced septicemia generally involves the use of mycobacterium-designated bottles, MGIT method, and Ogawa medium; however, we used microbe cultures with routine blood-culture bottles in the present case. Of the 24 mycobacterium-induced septicemia cases reported in the past 10 years, only eight cases were detected from routine blood-culture bottles; they were all rapidly growing bacteria. Mycobacteria other than the rapidly growing mycobacteria display delayed culture proliferation, therefore it is possible that non-detected microbes were probably present in the patients despite the fact that they were suffering from septicemia. In cases suspected to have severe infections, particularly those with a depressed immunodefense system, blood-culture testing for mycobacteria would be highly helpful for diagnosis.

  9. Effect of cord blood serum on ex vivo human limbal epithelial cell culture.

    PubMed

    Chakraborty, Anindita; Dutta, Jayanta; Das, Sumantra; Datta, Himadri

    2012-12-01

    Limbal cell transplantation is an efficacious procedure for rehabilitation of visual acuity in patients with severe ocular surface disorders. Cultivation of limbal epithelial stem cell with fetal bovine serum for transplantation has been a promising treatment for reconstructing the ocular surface in severe limbal stem cell deficiency caused by Steven Johnson syndrome, chemical or thermal injury. This technique of "cell therapy" has been accepted worldwide but the cost of cultivating the cells for transplantation is high. The objective of this study was to investigate the effect of cord blood serum in place of fetal bovine serum on the growth of human limbal epithelial cell culture. Our group has experimented with human cord blood serum which was obtained free of cost from willing donors. The use of human cord blood serum in place of fetal bovine serum for ex vivo culture of limbal stem cell has helped us in reducing the cost of culture. Fresh human limbal tissues from donor cadavers were cultured on intact and denuded amniotic membrane. Cells were proliferated in vitro with cell culture media containing human cord blood serum. Reverse transcription-polymerase chain reaction and immunofluorescence cytochemistry of cultured human limbal epithelial stem cell was done for characterization of the cells.

  10. Enzyme capture assay for rapid identification of Escherichia coli in blood cultures.

    PubMed Central

    Huang, S W; Wu, J J; Chang, T C

    1994-01-01

    An enzyme capture assay (ECA) for rapid identification of Escherichia coli in blood cultures by using beta-D-glucuronidase as a marker was developed. Microdilution plates coated with antiglucuronidase were used to capture this enzyme from the cell lysates of blood cultures which showed growth of gram-negative bacteria. The assay, using 4-methylumbelliferyl-beta-D-glucuronide as a fluorogenic substrate, had a detection limit of 0.1 ng/ml (3 x 10(-13) M) for the enzyme; this was approximately equal to a cell concentration of 10(6) CFU of E. coli per ml. Among 212 blood cultures showing growth of gram-negative bacteria, 77 specimens were found to contain E. coli by conventional culture procedures and 73 samples were positive by ECA. Among the 135 blood cultures from which E. coli was not isolated, ECA gave one false-positive (Salmonella enteritidis) reaction. Thus, the sensitivity and specificity for the identification of E. coli in blood cultures by ECA were 94.8% (73/77) and 99.3% (134/135), respectively. From the finding of positive growth in the culture bottle, the assay can be completed within 4 h. In view of the high rate of isolation of E. coli from bacteremic patients, the test can be performed in parallel with conventional culture protocols; this may shorten the identification time for E. coli, and proper antimicrobial treatments may be started 24 h earlier than when results of conventional identification systems are used. PMID:8077387

  11. A Multicenter Evaluation of Blood Culture Practices, Contamination Rates, and the Distribution of Causative Bacteria.

    PubMed

    Altindis, Mustafa; Koroglu, Mehmet; Demiray, Tayfur; Dal, Tuba; Ozdemir, Mehmet; Sengil, Ahmet Zeki; Atasoy, Ali Riza; Doğan, Metin; Cicek, Aysegul Copur; Ece, Gulfem; Kaya, Selcuk; Iraz, Meryem; Gultepe, Bilge Sumbul; Temiz, Hakan; Kandemir, Idris; Aksaray, Sebahat; Cetinkol, Yeliz; Sahin, Idris; Guducuoglu, Huseyin; Kilic, Abdullah; Kocoglu, Esra; Gulhan, Baris; Karabay, Oguz

    2016-01-01

    The prognostic value of blood culture testing in the diagnosis of bacteremia is limited by contamination. In this multicenter study, the aim was to evaluate the contamination rates of blood cultures as well as the parameters that affect the culture results. Sample collection practices and culture data obtained from 16 university/research hospitals were retrospectively evaluated. A total of 214,340 blood samples from 43,254 patients admitted to the centers in 2013 were included in this study. The blood culture results were evaluated based on the three phases of laboratory testing: the pre-analytic, the analytic, and the post-analytic phase. Blood samples were obtained from the patients through either the peripheral venous route (64%) or an intravascular catheter (36%). Povidone-iodine (60%) or alcohol (40%) was applied to disinfect the skin. Of the 16 centers, 62.5% have no dedicated phlebotomy team, 68.7% employed a blood culture system, 86.7% conducted additional studies with pediatric bottles, and 43.7% with anaerobic bottles. One center maintained a blood culture quality control study. The average growth rate in the bottles of blood cultures during the defined period (1259 - 26,400/year) was 32.3%. Of the growing microorganisms, 67% were causative agents, while 33% were contaminants. The contamination rates of the centers ranged from 1% to 17%. The average growth time for the causative bacteria was 21.4 hours, while it was 36.3 hours for the contaminant bacteria. The most commonly isolated pathogens were Escherichia coli (22.45%) and coagulase-negative staphylococci (CoNS) (20.11%). Further, the most frequently identified contaminant bacteria were CoNS (44.04%). The high contamination rates were remarkable in this study. We suggest that the hospitals' staff should be better trained in blood sample collection and processing. Sterile glove usage, alcohol usage for disinfection, the presence of a phlebotomy team, and quality control studies may all contribute to

  12. A Multicenter Evaluation of Blood Culture Practices, Contamination Rates, and the Distribution of Causative Bacteria

    PubMed Central

    Altindis, Mustafa; Koroglu, Mehmet; Demiray, Tayfur; Dal, Tuba; Ozdemir, Mehmet; Sengil, Ahmet Zeki; Atasoy, Ali Riza; Doğan, Metin; Cicek, Aysegul Copur; Ece, Gulfem; Kaya, Selcuk; Iraz, Meryem; Gultepe, Bilge Sumbul; Temiz, Hakan; Kandemir, Idris; Aksaray, Sebahat; Cetinkol, Yeliz; Sahin, Idris; Guducuoglu, Huseyin; Kilic, Abdullah; Kocoglu, Esra; Gulhan, Baris; Karabay, Oguz

    2016-01-01

    Background: The prognostic value of blood culture testing in the diagnosis of bacteremia is limited by contamination. Objectives: In this multicenter study, the aim was to evaluate the contamination rates of blood cultures as well as the parameters that affect the culture results. Materials and Methods: Sample collection practices and culture data obtained from 16 university/research hospitals were retrospectively evaluated. A total of 214,340 blood samples from 43,254 patients admitted to the centers in 2013 were included in this study. The blood culture results were evaluated based on the three phases of laboratory testing: the pre-analytic, the analytic, and the post-analytic phase. Results: Blood samples were obtained from the patients through either the peripheral venous route (64%) or an intravascular catheter (36%). Povidone-iodine (60%) or alcohol (40%) was applied to disinfect the skin. Of the 16 centers, 62.5% have no dedicated phlebotomy team, 68.7% employed a blood culture system, 86.7% conducted additional studies with pediatric bottles, and 43.7% with anaerobic bottles. One center maintained a blood culture quality control study. The average growth rate in the bottles of blood cultures during the defined period (1259 - 26,400/year) was 32.3%. Of the growing microorganisms, 67% were causative agents, while 33% were contaminants. The contamination rates of the centers ranged from 1% to 17%. The average growth time for the causative bacteria was 21.4 hours, while it was 36.3 hours for the contaminant bacteria. The most commonly isolated pathogens were Escherichia coli (22.45%) and coagulase-negative staphylococci (CoNS) (20.11%). Further, the most frequently identified contaminant bacteria were CoNS (44.04%). Conclusions: The high contamination rates were remarkable in this study. We suggest that the hospitals’ staff should be better trained in blood sample collection and processing. Sterile glove usage, alcohol usage for disinfection, the presence of

  13. [Confusion Over the Term "Contamination Rate" as It Pertains to Blood Cultures].

    PubMed

    Morii, Daiichi; Yokozawa, Takayuki; Ichinose, Naoki; Oda, Toshimi

    2016-05-01

    The blood culture contamination rate is often used to validate specimen-collection procedures. CUMITECH has set its optimal target to be 2% to 3%. However, the term "contamination rate" has been defined in many ways, limiting its generalizability. The definitions used in earlier studies can be divided into two categories; definitions based on clinical judgements, and those based on preset rules. According to each principle, the equation must be composed of a defined numerator and denominator. The problem with clinical definitions is that the decision is inevitably subjective, and the process is too cumbersome. Also, if the number of positive cultures is used as the denominator, the value would be equivalent to the positive predictive value, given that contamination is regarded as a "positive case." Thus, the value would not be useful for validating a procedure. On the other hand, when the preset algorithm was adopted, true infection would, to some degree, inevitably be classified as contamination. Also, if the algorithm adopted the number of blood culture sets as the denominator and contamination was defined as the identification of 1 or more specified organisms in only 1 of multiple sets of blood cultures, its theoretical maximum value would not be 100%. This is a problem because the value is a mixture of several numbers with different scales. In other words, whether the blood cultures are collected once, twice, or thrice or more a day would affect the result. The study cited by CUMITECH aimed to evaluate the equivalence between the clinical definition and the laboratory definition with preset rules, rather than to establish a benchmark for the contamination rate. It is undesirable for the number to be perceived as a benchmark. "A Guide to Blood Culture" (2013) by the Japanese Society for Clinical Microbiology introduced a calculation for the contamination rate, but the definition of the term "number of specimens" in the formula is ambiguous. In addition, the

  14. Blood culture-based diagnosis of bacteraemia: state of the art.

    PubMed

    Opota, O; Croxatto, A; Prod'hom, G; Greub, G

    2015-04-01

    Blood culture remains the best approach to identify the incriminating microorganisms when a bloodstream infection is suspected, and to guarantee that the antimicrobial treatment is adequate. Major improvements have been made in the last years to increase the sensitivity and specificity and to reduce the time to identification of microorganisms recovered from blood cultures. Among other factors, the introduction in clinical microbiology laboratories of the matrix-assisted laser desorption ionization time-of-flight mass spectrometry technology revolutionized the identification of microorganisms whereas the introduction of nucleic-acid-based methods, such as DNA hybridization or rapid PCR-based test, significantly reduce the time to results. Together with traditional antimicrobial susceptibility testing, new rapid methods for the detection of resistance mechanisms respond to major epidemiological concerns such as methicillin-resistant Staphylococcus aureus, extended-spectrum β-lactamase or carbapenemases. This review presents and discusses the recent developments in microbial diagnosis of bloodstream infections based on blood cultures.

  15. Influence of postmortem time on the outcome of blood cultures among cadaveric tissue donors.

    PubMed

    Saegeman, V; Verhaegen, J; Lismont, D; Verduyckt, B; De Rijdt, T; Ectors, N

    2009-02-01

    Tissue banks provide tissues of human cadaver donors for transplantation. The maximal time limit for tissue retrieval has been set at 24 h postmortem. This study aimed at evaluating the evidence for this limit from a microbiological point of view. The delay of growth in postmortem blood cultures, the identification of the species isolated and clinical/environmental factors were investigated among 100 potential tissue donors. No significant difference was found in the rate of donors with grown blood cultures within (25/65=38%) compared with after (24/65=37%) 24 h of death. Coagulase-negative staphylococci and gastro-intestinal microorganisms were isolated within and after 24 h of death. Two factors--antimicrobial therapy and "delay before body cooling"--were significantly inversely related with donors' blood culture results. From a microbiological point of view, there is no evidence for avoiding tissue retrieval among donors after 24 h of death.

  16. Preliminary evaluation of a new clinical algorithm to interpret blood cultures growing coagulase-negative staphylococci.

    PubMed

    Schnell, David; Lécuyer, Hervé; Geeraerts, Thomas; Dumenil, Anne-Sylvie; Bille, Emmanuelle; Mercier, Frédéric J; Benhamou, Dan; Zahar, Jean-Ralph

    2013-07-01

    Evaluating the clinical significance of blood cultures positive for coagulase-negative staphylococci (CoNS) is of critical importance since these microorganisms represent both the first contaminants of blood cultures and one of the leading causes of bloodstream infection (BSI). This prospective 2-centre study aimed to compare a previously reported algorithm to a clinical algorithm based on our experience. We identified 84 patients with CoNS-positive blood cultures. Twenty-seven (32%) were considered to have BSI according to our study algorithm. Thirty-seven (44%) patients were considered to have CoNS BSI according to the previously reported algorithm. The 2 algorithms isolated patients with similar rates of recurrences and hospital mortality. Our algorithm seemed to result in less diagnoses of CoNS BSI without harmful consequences compared to the previously reported algorithm. The impact on patient outcome and the inappropriate use of antibiotics deserves further investigation.

  17. Blood culture sampling rates at a German pediatric university hospital and incidence of invasive pneumococcal disease.

    PubMed

    Rüggeberg, J U; Ketteler, K; MacKenzie, C R; Von Kries, R; Reinert, R R; Schroten, H

    2004-04-01

    Recent pediatric surveillance studies suggest the incidence of pneumococcal bacteremia, but not meningitis, is lower in Germany than in most developed countries. Suboptimal case assessment in routine clinical practice has been suspected of contributing to this apparent discrepancy. We aimed to assess the blood culture sampling rate at a German pediatric university hospital and the disease burden associated with pneumococcal bacteremia in children under 5 years of age. The study design was retrospective, based on data-linkage and chart review. Blood cultures were frequently obtained in sepsis (96%; CI 78-99%) and meningitis (95%; CI 77-99%), but less commonly in pneumonia (49%; CI 43-54%) and fever without focus (48%; CI 38-59%). Pneumococci were the most common source of clinically significant bacteremia in previously healthy children. These blood culture sampling rates may be insufficient for the sensitive detection of pneumococcal bacteremia. Epidemiological surveillance based on poorly standardized diagnostic practices is prone to under-assessment.

  18. Oxidatively damaged guanosine in white blood cells and in urine of welders: associations with exposure to welding fumes and body iron stores.

    PubMed

    Pesch, Beate; Lotz, Anne; Koch, Holger M; Marczynski, Boleslaw; Casjens, Swaantje; Käfferlein, Heiko U; Welge, Peter; Lehnert, Martin; Heinze, Evelyn; Van Gelder, Rainer; Hahn, Jens-Uwe; Behrens, Thomas; Raulf, Monika; Hartwig, Andrea; Weiss, Tobias; Brüning, Thomas

    2015-08-01

    The International Agency for Research on Cancer considers the carcinogenicity of welding fume of priority for re-evaluation. Genotoxic effects in experimental animals are still inconclusive. Here, we investigated the association of personal exposure to metals in respirable welding fumes during a working shift with oxidatively damaged guanosine in DNA of white blood cells (WBC) and in postshift urine samples from 238 welders. Medians of 8-oxo-7,8-dihydro-2'-deoxyguanosine (8-oxodGuo) were 2.35/10(6) dGuo in DNA of WBC and 4.33 µg/g creatinine in urine. The median of 8-oxo-7,8-dihydroguanosine (8-oxoGuo) was 7.03 µg/g creatinine in urine. The extent of both urinary parameters was higher in welders applying techniques with high particle emission rates to stainless steel than in tungsten inert gas welders (8-oxodGuo: 9.96 vs. 4.49 µg/L, 8-oxoGuo: 15.7 vs. 7.7 µg/L), but this apparent difference diminished after creatinine adjustment. We applied random intercept models to estimate the influence of airborne and systemic exposure to metals on oxidatively damaged guanosine in WBC and urine together with covariates. We observed a highly significant nonlinear association of urinary 8-oxoGuo with serum ferritin (P < 0.0001) and higher 8-oxoGuo concentrations for respirable iron >1,000 µg/m(3) compared to ≤57 µg/m(3). Similar effects were found for manganese. Airborne chromium but not nickel was associated with all oxidatively modified guanosine measures, whereas urinary chromium as well as nickel showed associations with urinary modified guanosines. In summary, oxidatively damaged urinary guanosine was associated with airborne and systemic exposure to metals in welders and showed a strong relation to body iron stores.

  19. Melatonin and IP3-induced Ca2+ Release from Intracellular Stores in the Malaria Parasite Plasmodium falciparum within Infected Red Blood Cells*

    PubMed Central

    Alves, Eduardo; Bartlett, Paula J.; Garcia, Celia R. S.; Thomas, Andrew P.

    2011-01-01

    IP3-dependent Ca2+ signaling controls a myriad of cellular processes in higher eukaryotes and similar signaling pathways are evolutionarily conserved in Plasmodium, the intracellular parasite that causes malaria. We have reported that isolated, permeabilized Plasmodium chabaudi, releases Ca2+ upon addition of exogenous IP3. In the present study, we investigated whether the IP3 signaling pathway operates in intact Plasmodium falciparum, the major disease-causing human malaria parasite. P. falciparum-infected red blood cells (RBCs) in the trophozoite stage were simultaneously loaded with the Ca2+ indicator Fluo-4/AM and caged-IP3. Photolytic release of IP3 elicited a transient Ca2+ increase in the cytosol of the intact parasite within the RBC. The intracellular Ca2+ pools of the parasite were selectively discharged, using thapsigargin to deplete endoplasmic reticulum (ER) Ca2+ and the antimalarial chloroquine to deplete Ca2+ from acidocalcisomes. These data show that the ER is the major IP3-sensitive Ca2+ store. Previous work has shown that the human host hormone melatonin regulates P. falciparum cell cycle via a Ca2+-dependent pathway. In the present study, we demonstrate that melatonin increases inositol-polyphosphate production in intact intraerythrocytic parasite. Moreover, the Ca2+ responses to melatonin and uncaging of IP3 were mutually exclusive in infected RBCs. Taken together these data provide evidence that melatonin activates PLC to generate IP3 and open ER-localized IP3-sensitive Ca2+ channels in P. falciparum. This receptor signaling pathway is likely to be involved in the regulation and synchronization of parasite cell cycle progression. PMID:21149448

  20. Blood culture contamination in hospitalized pediatric patients: a single institution experience

    PubMed Central

    Min, Hyewon; Park, Cheong Soo; Kim, Dong Soo

    2014-01-01

    Purpose Blood culture is the most important tool for detecting bacteremia in children with fever. However, blood culture contamination rates range from 0.6% to 6.0% in adults; rates for young children have been considered higher than these, although data are limited, especially in Korea. This study determined the contamination rate and risk factors in pediatric patients visiting the emergency room (ER) or being admitted to the ward. Methods We conducted a retrospective chart review of blood cultures obtained from children who visited Yonsei Severance Hospital, Korea between 2006 and 2010. Positive blood cultures were labeled as true bacteremia or contamination according to Centers for Disease Control and Prevention/National Healthcare Safety Network definitions for laboratory-confirmed bloodstream infection, after exclusion of cultures drawn from preexisting central lines only. Results Among 40,542 blood cultures, 610 were positive, of which 479 were contaminations and 131 were true bacteremia (overall contamination rate, 1.18%). The contamination rate in the ER was significantly higher than in the ward (1.32% vs. 0.66%, P<0.001). The rate was higher in younger children (2.07%, 0.94%, and 0.61% in children aged <1 year, 1-6 years, and >6 years, respectively). Conclusion Overall, contamination rates were higher in younger children than in older children, given the difficulty of performing blood sampling in younger children. The contamination rates from the ER were higher than those from the ward, not accounted for only by overcrowding and lack of experience among personnel collecting samples. Further study to investigate other factors affecting contamination should be required. PMID:24868215

  1. [Quality indicators for blood culture: three years of monitoring at a university hospital in Chile].

    PubMed

    Guzmán, Ana María; Sánchez, Tomás; de la Barra, Ricardo

    2012-08-01

    Blood culture is considered the "gold standard" for the diagnosis of bacteremia, critical condition with high morbidity and mortality. Because of its importance, it is estimated that the blood culture is a critical test that requires close monitoring on the quality with which the process is performed. The objective of this work is to show the results of the monitoring carried out during the past three years, of 5 quality indicators of blood cultures in the laboratory of the Hospital Clínico de la Pontificia Universidad Católica de Chile, considering pre-analytical, analytical and post-analytical aspects. In the 3 years monitored the mean contamination was 0,7%, 46% of adult bottles had adequate volume, match between Gram stain with final identification was 99.4%, 100% of correct participations were achieved in surveys of external quality control and Gram staining notification before 1 hour was 88.7%. With regard to proposed aims, in 2011 the laboratory complies with all, except the percentage of bottles with appropriate volume of blood inoculated. This indicator is very low and should be corrected as soon as possible since it is known that it is an important condition for optimum performance of blood cultures.

  2. Validation of a monoclonal antibody-based immunofluorescent assay to detect Burkholderia pseudomallei in blood cultures

    PubMed Central

    Dulsuk, Adul; Paksanont, Suporn; Sangchankoom, Adisak; Ekchariyawat, Peeraya; Phunpang, Rungnapa; Jutrakul, Yaowaruk; West, T. Eoin

    2016-01-01

    Abstract Background Identification of Burkholderia pseudomallei, the cause of melioidosis, using routine methods takes several days. Use of a monoclonal antibody-based immunofluorescent assay (IFA) on positive blood cultures may speed diagnosis. Methods We tested the diagnostic accuracy of the IFA on 545 blood cultures positive for Gram-negative organisms at Udon Thani Hospital, Thailand, between June 2015 and August 2016. Results Sensitivity of the IFA was 100% and specificity was 99.6%. The median decrease in time to pathogen identification between the IFA result and routine methods was 28 h (IQR 25–51), p<0.0001. Conclusions The IFA accurately expedites the diagnosis of melioidosis. PMID:28115683

  3. Large-scale clinical comparison of the lysis-centrifugation and radiometric systems for blood culture

    SciTech Connect

    Brannon, P.; Kiehn, T.E.

    1985-12-01

    The Isolator 10 lysis-centrifugation blood culture system (E. I. du Pont de Nemours and Co., Inc., Wilmington, Del.) was compared with the BACTEC radiometric method (Johnston Laboratories, Inc., Towson, Md.) with 6B and 7D broth media for the recovery of bacteria and yeasts. From 11,000 blood cultures, 1,174 clinically significant organisms were isolated. The Isolator system recovered significantly more total organisms, members of the family Enterobacteriaceae, Staphylococcus spp., and yeasts. The BACTEC system recovered significantly more Pseudomonas spp., Streptococcus spp., and anaerobes. Of the Isolator colony counts, 87% measured less than 11 CFU/ml of blood. Organisms, on an average, were detected the same day from each of the two culture systems. Only 13 of the 975 BACTEC isolates (0.01%) were recovered by subculture of growth-index-negative bottles, and 12 of the 13 were detected in another broth blood culture taken within 24 h. Contaminants were recovered from 4.8% of the Isolator 10 and 2.3% of the BACTEC cultures.

  4. Relevance of blood cultures in acute pyelonephritis in a single-center retrospective study.

    PubMed

    Ledochowski, Stanislas; Abraham, Paul-Samuel; Jacob, Xavier; Dumitrescu, Oana; Lina, Gérard; Lepape, Alain; Piriou, Vincent; Wallet, Florent; Friggeri, Arnaud

    2015-08-01

    Pyelonephritides are frequently encountered diagnosis in Emergency Departments. Urinalyses have a central place in the management of this situation but the usefulness of blood cultures is not clear. We conducted a single-center retrospective study of 24 months to study the microbiological relevance of blood cultures in pyelonephritis. We included patients with blood cultures (BC) and urine cultures (UC) drawn at the same time, if they were not exposed to antibiotics prior to these tests. Of our 264 patients, 39 (15 %) had no bacteriological documentation. There were 83 (31 %) bacteremic patients. Seven patients had contaminated or sterile UC with positive BC. Four patients had positive UC and BC with the latter allowing identification of a pathogen absent from the UC (n = 1) or identifying the main pathogen in three cases. A total of 11 patients theoretically benefited from BC representing 4.2 % of our population. Excluding one patient who was known to be infected with multi-drug resistant bacteria, all empirical antibiotics regimens were effective against the identified pathogens. We did not reveal any significant therapeutic impact of blood cultures in the management of pyelonephritis, when BC and UC are performed before any antimicrobials treatment.

  5. Culture and characterisation of equine peripheral blood mesenchymal stromal cells.

    PubMed

    Spaas, Jan H; De Schauwer, Catharina; Cornillie, Pieter; Meyer, Evelyne; Van Soom, Ann; Van de Walle, Gerlinde R

    2013-01-01

    Although the use of mesenchymal stromal cells (MSCs) for the treatment of orthopaedic injuries in horses has been reported, no official guidelines exist that classify a particular cell as an equine MSC. Given the limited characterisation of peripheral blood (PB)-derived equine MSCs in particular, this study aimed to provide more detailed information in relation to this cell type. Mesenchymal stromal cells were isolated from equine PB samples and colony forming unit (CFU) assays as well as population doubling times (PDTs) (from P(0) to P(10)) were performed. Two types of colonies, 'fingerprint' and dispersed, could be observed based on macroscopic and microscopic features. Moreover, after an initial lag phase (as indicated by a negative PDT at P(0) to P(1)) the MSCs divided rapidly as indicated by a positive PDT at all further passages. Immunophenotyping was carried out with trypsin- as well as with accutase-detached MSC to evaluate potential trypsin-sensitive epitope destruction on particular antigens. Isolated MSC were positive for CD29, CD44, CD90 and CD105, and negative for CD45, CD79α, MHC II and a monocyte/macrophage marker, irrespective of the cell detaching agent used. Trilineage differentiation of the MSCs towards osteoblasts, chondroblasts and adipocytes was confirmed using a range of histochemical stains. Copyright © 2012 Elsevier Ltd. All rights reserved.

  6. Use of BBL CHROMagar MRSA Medium for Identification of Methicillin-Resistant Staphylococcus aureus Directly from Blood Cultures

    PubMed Central

    Pape, John; Wadlin, Jill; Nachamkin, Irving

    2006-01-01

    We evaluated the ability of BBL CHROMagar MRSA medium (Becton Dickinson, Sparks, MD) to identify methicillin-resistant Staphylococcus aureus (MRSA) directly upon subculture from positive blood culture bottles. There were 124 MRSA isolates recovered from blood cultures in the study. BBL CHROMagar MRSA medium was highly sensitive (97.6% [121/124] at 18 to 24 h of incubation and 100% [124/124] at 48 h) and 99.9% specific for identifying MRSA from positive blood cultures. PMID:16825383

  7. Faster identification of pathogens in positive blood cultures by fluorescence in situ hybridization in routine practice.

    PubMed

    Peters, Remco P H; Savelkoul, Paul H M; Simoons-Smit, Alberdina M; Danner, Sven A; Vandenbroucke-Grauls, Christina M J E; van Agtmael, Michiel A

    2006-01-01

    Rapid identification of microorganisms in blood cultures is required to optimize empirical treatment at an early stage. Fluorescence in situ hybridization (FISH) can reduce the time to identification of microorganisms in growth-positive blood cultures. In this study, we evaluated the performance, time to identification, and potential clinical benefits of FISH compared to those of conventional culture methods in routine practice. After Gram staining, blood culture fluids were simultaneously further identified with FISH and with conventional culture methods. Results and points in time of FISH and culture identification (provisional and final identifications) were collected and compared. For 91% of microorganisms, the genus or family name was identified, and for 79%, the species name could be attributed. The sensitivity and specificity of the individual probes exceeded 95%, except for the Enterobacteriaceae probe (sensitivity, 89%). Cross-hybridization was obtained with the Klebsiella pneumoniae probe for Klebsiella oxytoca. The time gains of FISH and final culture identification were more than 18 h for bacteria and 42 h for yeasts. With FISH, Staphylococcus aureus was differentiated from coagulase-negative staphylococci 1.4 h faster than by provisional identification (P < 0.001). In conclusion, FISH allows rapid and reliable identification of the majority of microorganisms in growth-positive blood cultures. The substantial time gain of identification with FISH may allow same-day adjustment of antimicrobial therapy, and FISH is especially useful if no provisional identification is obtained. With further extension of the number of probes and a reduction in turnaround time, FISH will become a very useful diagnostic tool in the diagnosis of bloodstream infections.

  8. A model of psychosocial and cultural antecedents of blood pressure control.

    PubMed Central

    Bosworth, Hayden B.; Oddone, Eugene Z.

    2002-01-01

    Hypertension is a major modifiable risk factor for stroke, congestive heart failure, and end-stage renal disease. Hypertension is particularly prevalent and deadly among African Americans. Effective treatment for hypertension has been available for decades, yet only one fourth of all individuals have their blood pressure under control. Despite the potential impact of hypertension, interventions to improve control have had limited success. We present a model of how to understand antecedents of blood pressure control according to three interrelated categories: patient characteristics, social and cultural environment, and medical environment. This theoretical paper was conducted using a literature review and a model to explain psychosocial antecedents of blood pressure control is presented. We conclude that improved understanding of important antecedents of blood pressure control coupled with technological advances, such as tailored interventions, provide clinicians with a tool that may lead to improved blood pressure control. These interventions will require the involvement of clinicians and consideration of sociocultural factors to be successful. PMID:11991336

  9. Measurement of Blood Coagulation Factor Synthesis in Cultures of Human Hepatocytes.

    PubMed

    Heinz, Stefan; Braspenning, Joris

    2015-01-01

    An important function of the liver is the synthesis and secretion of blood coagulation factors. Within the liver, hepatocytes are involved in the synthesis of most blood coagulation factors, such as fibrinogen, prothrombin, factor V, VII, IX, X, XI, XII, as well as protein C and S, and antithrombin, whereas liver sinusoidal endothelial cells produce factor VIII and von Willebrand factor. Here, we describe methods for the detection and quantification of most blood coagulation factors in hepatocytes in vitro. Hepatocyte cultures indeed provide a valuable tool to study blood coagulation factors. In addition, the generation and expansion of hepatocytes or hepatocyte-like cells may be used in future for cell-based therapies of liver diseases, including blood coagulation factor deficiencies.

  10. PCR identification of bacteria in blood culture does not fit the daily workflow of a routine microbiology laboratory.

    PubMed

    Karumaa, Santra; Kärpänoja, Pauliina; Sarkkinen, Hannu

    2012-03-01

    We have evaluated the GenoType blood culture assay (Hain Lifescience, Nehren, Germany) for the identification of bacteria in 233 positive blood cultures and assessed its suitability in the workflow of a routine microbiology laboratory. In 68/233 (29.2%) samples, the culture result could not be confirmed by the GenoType assay due to a lack of primers in the test, multiple organisms in the sample, or inconsistency with respect to the identification by culture. Although the GenoType blood culture assay gives satisfactory results for bacteria for which primers are available, there are difficulties in applying the test in the routine microbiology laboratory.

  11. Tobacco advertising in retail stores.

    PubMed Central

    Cummings, K M; Sciandra, R; Lawrence, J

    1991-01-01

    Recent studies have described tobacco advertising in the print media, on billboards, and through sponsorship of cultural and sporting events. However, little attention has been given to another common and unavoidable source of tobacco advertising, that which is encountered in retail stores. In July 1987, we conducted a survey of 61 packaged goods retail stores in Buffalo, NY, to assess the prevalence and type of point-of-sale tobacco advertising. In addition, store owners or managers were surveyed to determine their store's policy regarding tobacco advertising, receipt of monetary incentives from distributors for displaying tobacco ads, and willingness to display antitobacco ads. Six types of stores were involved in the study: 10 supermarkets, 10 privately owned grocery stores, 9 chain convenience food stores that do not sell gasoline, 11 chain convenience food stores that sell gasoline, 11 chain pharmacies, and 10 private pharmacies. Two-thirds of the stores displayed tobacco posters, and 87 percent had promotional items advertising tobacco products, primarily cigarettes. Larger stores, and those that were privately owned, tended to display more posters and promotional items. Eighty percent of tobacco product displays were for cigarettes, 16 percent for smokeless tobacco products, and 4 percent for cigars and pipe tobacco. Convenience stores selling gasoline had the most separate tobacco product displays. Of tobacco product displays, 24 percent were located adjacent to candy and snack displays. Twenty-nine of the 61 store owners or managers indicated that their store had a policy regulating the display of tobacco ads and tobacco product displays.(ABSTRACT TRUNCATED AT 250 WORDS) PMID:1910192

  12. One size doesn’t fit all: Should we reconsider the introduction of cold-stored platelets in blood bank inventories?

    PubMed Central

    Berzuini, Alessandra; Spreafico, Marta; Prati, Daniele

    2017-01-01

    Platelet concentrates are universally prepared with a standard method and stored for 5 days at room temperature (20–24°C) in gentle agitation. Currently, there is a renewed interest in the possibility of storing platelet concentrates below the standard temperatures. In fact, cold platelets might be more effective in bleeding patients and have a lower risk of bacterial transmission. Inventories including platelets at different temperatures may favour patient-centred strategies for prophylactic or therapeutic transfusions. PMID:28184297

  13. One size doesn't fit all: Should we reconsider the introduction of cold-stored platelets in blood bank inventories?

    PubMed

    Berzuini, Alessandra; Spreafico, Marta; Prati, Daniele

    2017-01-01

    Platelet concentrates are universally prepared with a standard method and stored for 5 days at room temperature (20-24°C) in gentle agitation. Currently, there is a renewed interest in the possibility of storing platelet concentrates below the standard temperatures. In fact, cold platelets might be more effective in bleeding patients and have a lower risk of bacterial transmission. Inventories including platelets at different temperatures may favour patient-centred strategies for prophylactic or therapeutic transfusions.

  14. Performance of Five Agar Media for Recovery of Fungi from Isolator Blood Cultures

    PubMed Central

    Procop, G. W.; Cockerill, F. R.; Vetter, E. A.; Harmsen, W. S.; Hughes, J. G.; Roberts, G. D.

    2000-01-01

    We studied the recovery of 1,270 fungal isolates from 176,144 Isolator blood cultures (0.72% positive) on bacterial and fungal media, under routine and differing incubation conditions. Except with Histoplasma capsulatum, chocolate agar incubated for only 3 days proved to be an excellent medium for the recovery of fungi from the Isolator system. PMID:11015411

  15. Rapid Intrinsic Fluorescence Method for Direct Identification of Pathogens in Blood Cultures

    PubMed Central

    Walsh, John D.; Hyman, Jay M.; Borzhemskaya, Larisa; Bowen, Ann; McKellar, Caroline; Ullery, Michael; Mathias, Erin; Ronsick, Christopher; Link, John; Wilson, Mark; Clay, Bradford; Robinson, Ron; Thorpe, Thurman; van Belkum, Alex; Dunne, W. Michael

    2013-01-01

    ABSTRACT A positive blood culture is a critical result that requires prompt identification of the causative agent. This article describes a simple method to identify microorganisms from positive blood culture broth within the time taken to perform a Gram stain (<20 min). The method is based on intrinsic fluorescence spectroscopy (IFS) of whole cells and required development of a selective lysis buffer, aqueous density cushion, optical microcentrifuge tube, and reference database. A total of 1,121 monomicrobial-positive broth samples from 751 strains were analyzed to build a database representing 37 of the most commonly encountered species in bloodstream infections or present as contaminants. A multistage algorithm correctly classified 99.6% of unknown samples to the Gram level, 99.3% to the family level, and 96.5% to the species level. There were no incorrect results given at the Gram or family classification levels, while 0.8% of results were discordant at the species level. In 8/9 incorrect species results, the misidentified isolate was assigned to a species of the same genus. This unique combination of selective lysis, density centrifugation, and IFS can rapidly identify the most common microbial species present in positive blood cultures. Faster identification of the etiologic agent may benefit the clinical management of sepsis. Further evaluation is now warranted to determine the performance of the method using clinical blood culture specimens. PMID:24255123

  16. Carbapenem resistance via the blaKPC-2 gene in Enterobacter cloacae blood culture isolate.

    PubMed

    Lo, Andy; Verrall, Rosemary; Williams, John; Stratton, Charles; Della-Latta, Phyllis; Tang, Yi-Wei

    2010-05-01

    An Enterobacter cloacae blood culture isolate expressing carbapenem resistance via the Klebsiella pneumoniae carbapenemase KPC-2 gene is reported. To our knowledge, this is the first report of a nosocomial isolate with carbapenemase-mediated resistance causing infection in a patient from Tennessee.

  17. Preparation of positive blood cultures for direct MALDI-ToF MS identification.

    PubMed

    Robinson, Andrew M; Ussher, James E

    2016-08-01

    MALDI-ToF MS can be used to identify microorganisms directly from blood cultures. This study compared two methods of sample preparation. Similar levels of genus- (91% vs 90%) and species-level identifications (79% vs 74%) were obtained with differential centrifugation and SDS methods. The SDS method is faster and requires minimal handling.

  18. [Rapid identification of microorganisms by mass spectrometry in a blood culture system. Comparison of two procedures].

    PubMed

    Cattani, María E; Posse, Tamara; Hermes, Ricardo L; Kaufman, Sara C

    2015-01-01

    Rapid identification of microorganisms is critical in hospitalized infected patients. Blood culture is currently the gold standard for detecting and identifying microorganisms causing bacteremia or sepsis. The introduction of mass spectrometry by matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF MS) in microbiology laboratories, especially in microorganisms growing in blood culture bottles, provides rapid identification. This study evaluates the performance of the Maldi Sepsityper Biotyper procedure (hereinafter, MS) compared to that of an in-home method (hereinafter, HF). Eight hundred and forty (840) positive blood culture bottles were processed using the HF procedure, 542 of which were also processed using MS. The organisms were identified in 670 (79.76%) and 391 (72.14%) bottles respectively (p = 0,0013). This study demonstrates the effectiveness of both procedures for identifying microorganisms directly from positive blood culture bottles. However, the HF procedure proved to be more effective than MS, especially in the presence of Gram positive organisms.

  19. Molecular Characterization of a Catalase-Negative Staphylococcus aureus Blood Culture Isolate

    PubMed Central

    Kum, Steven; Jureen, Roland; Lin, Raymond T. P.

    2015-01-01

    Here we report a catalase-negative methicillin-sensitive Staphylococcus aureus isolate collected from a blood culture. Sequencing through the gene encoding catalase, katA, demonstrated a 2-bp insertion. The resulting frameshift mutation generates a protein that has lost 26 amino acids (aa) at its C-terminal domain. PMID:26354811

  20. Predicting bacteremia based on nurse-assessed food consumption at the time of blood culture.

    PubMed

    Komatsu, Takayuki; Onda, Toshihito; Murayama, Go; Yamanouchi, Masashi; Inukai, Minori; Sakai, Ai; Kikuta, Masumi; Branch, Joel; Aoki, Makoto; Tierney, Lawrence M; Inoue, Kenji

    2012-01-01

    Bacteremia and its complications are important causes of morbidity and mortality in hospitalized patients. However, the yield of blood cultures is relatively low, with many false-positive results from bacterial contamination. We investigated the relationship between patient food consumption and the presence of bacteremia. This was an observational analysis of a cohort of 1179 patients who underwent blood culture analysis between January 2005 and December 2009. Patients with anorexia-inducing conditions, such as gastrointestinal illness and malignant disease treated with chemotherapy, were excluded. Food consumption was rated by nurses as the percentage of food consumed during the meal preceding the blood culture. Groupings were as follows: low consumption (<50%), moderate (>50% to <80%), and high (>80%). Low consumption was observed in 39.8% of patients, moderate in 17.8%, and high in 41.6%. The average body temperature was 38.1 ± 1.1°C. Bacteremia was present in 18.5%, 3.9%, and 1.4% of patients in the low, moderate, and high food consumption groups, respectively. The negative predictive value was 98.3%, suggesting that bacteremia is very unlikely in the setting of good food intake. Bacteremia is an unlikely occurrence in hospitalized patients who maintain adequate food consumption at the time of blood culture. Copyright © 2012 Society of Hospital Medicine.

  1. Western Culture in Japanese Film: Kurosawa's "Throne of Blood" and "Ran."

    ERIC Educational Resources Information Center

    Kane, Peter E.

    Akira Kurosawa, the most popular Asian film maker with audiences in the United States, has found in William Shakespeare's plays themes and plots that resonate within Japanese culture. While the translations of "Macbeth" into "Throne of Blood" and "King Lear" into "Ran" are quite direct and literal with only…

  2. Western Culture in Japanese Film: Kurosawa's "Throne of Blood" and "Ran."

    ERIC Educational Resources Information Center

    Kane, Peter E.

    Akira Kurosawa, the most popular Asian film maker with audiences in the United States, has found in William Shakespeare's plays themes and plots that resonate within Japanese culture. While the translations of "Macbeth" into "Throne of Blood" and "King Lear" into "Ran" are quite direct and literal with only…

  3. Comparative usefulness of inflammatory markers to indicate bacterial infection-analyzed according to blood culture results and related clinical factors.

    PubMed

    Nishikawa, Hirokazu; Shirano, Michinori; Kasamatsu, Yu; Morimura, Ayumi; Iida, Ko; Kishi, Tomomi; Goto, Tetsushi; Okamoto, Saki; Ehara, Eiji

    2016-01-01

    To assess relationships of inflammatory markers and 2 related clinical factors with blood culture results, we retrospectively investigated inpatients' blood culture and blood chemistry findings that were recorded from January to December 2014 using electronic medical records and analyzed the data of 852 subjects (426 culture-positive and 426 culture-negative). Results suggested that the risk of positive blood culture statistically increased as inflammatory marker levels and the number of related factors increased. Concerning the effectiveness of inflammatory markers, when the outcome definition was also changed for C-reactive protein (CRP), the odds ratio had a similar value, whereas when the outcome definition of blood culture positivity was used for procalcitonin (PCT), the greatest effectiveness of that was detected. Therefore, the current results suggest that PCT is more useful than CRP as an auxiliary indication of bacterial infection.

  4. Cultural diversity as a factor in self-monitoring blood glucose in gestational diabetes.

    PubMed

    Langer, O; Langer, N; Piper, J M; Elliott, B; Anyaegbunam, A

    1995-01-01

    The routine use of self-monitoring of capillary blood glucose by pregnant diabetic patients currently provides the basis for both clinical management and ongoing investigation. Strategies must therefore be developed to ensure that these data are reliable and accurately reported by patients and are not influenced by diverse socioeconomic levels or varied geographic locations. To explore this issue, we used glucose reflectance meters with a memory microchip capable of storing up to 440 consecutive blood glucose determinations. Two diverse groups of women from Texas and New York who had gestational diabetes performed self-monitoring of blood glucose from diagnosis until delivery. Both groups recorded their blood glucose results daily in a logbook. The reporting performance of all the participating subjects resulted in an actual compliance rate of 60% to 70% of testings required of the patients. Comparison of African-American, Mexican-American, and white populations revealed no significant differences in patient performance or compliance. Moreover, no differences were found between the groups at different geographic locations (New York, Texas) in patients' willingness and ability to comply with the regimen of self-monitoring blood glucose. These findings suggest that the use of memory reflectance meters, in conjunction with patient education and positive interaction between patient and care provider, will result in high patient compliance regardless of socioeconomic level or ethnic diversity.

  5. Multiplex blood PCR in combination with blood cultures for improvement of microbiological documentation of infection in febrile neutropenia.

    PubMed

    Lamoth, F; Jaton, K; Prod'hom, G; Senn, L; Bille, J; Calandra, T; Marchetti, O

    2010-10-01

    The frequent lack of microbiological documentation of infection by blood cultures (BC) has a major impact on clinical management of febrile neutropenic patients, especially in cases of unexplained persistent fever. We assessed the diagnostic utility of the LightCycler SeptiFast test (SF), a multiplex blood PCR, in febrile neutropenia. Blood for BC and SF was drawn at the onset of fever and every 3 days of persistent fever. SF results were compared with those of BC, clinical documentation of infection, and standard clinical, radiological, and microbiological criteria for invasive fungal infections (IFI). A total of 141 febrile neutropenic episodes in 86 hematological patients were studied: 44 (31%) microbiologically and 49 (35%) clinically documented infections and 48 (34%) unexplained fevers. At the onset of fever, BC detected 44 microorganisms in 35/141 (25%) episodes. Together, BC and SF identified 78 microorganisms in 61/141 (43%) episodes (P = 0.002 versus BC or SF alone): 12 were detected by BC and SF, 32 by BC only, and 34 by SF only. In 19/52 (37%) episodes of persistent fever, SF detected 28 new microorganisms (7 Gram-positive bacterial species, 15 Gram-negative bacterial species, and 6 fungal species [89% with a clinically documented site of infection]) whereas BC detected only 4 pathogens (8%) (P = 0.001). While BC did not detect fungi, SF identified 5 Candida spp. and 1 Aspergillus sp. in 5/7 probable or possible cases of IFI. Using SeptiFast PCR combined with blood cultures improves microbiological documentation in febrile neutropenia, especially when fever persists and invasive fungal infection is suspected. Technical adjustments may enhance the efficiency of this new molecular tool in this specific setting.

  6. Measurement of Microbial DNA Polymerase Activity Enables Detection and Growth Monitoring of Microbes from Clinical Blood Cultures

    PubMed Central

    Zweitzig, Daniel R.; Riccardello, Nichol M.; Morrison, John; Rubino, Jason; Axelband, Jennifer; Jeanmonod, Rebecca; Sodowich, Bruce I.; Kopnitsky, Mark J.; O’Hara, S. Mark

    2013-01-01

    Surveillance of bloodstream infections (BSI) is a high priority within the hospital setting. Broth-based blood cultures are the current gold standard for detecting BSI, however they can require lengthy incubation periods prior to detection of positive samples. We set out to demonstrate the feasibility of using enzymatic template generation and amplification (ETGA)-mediated measurement of DNA polymerase activity to detect microbes from clinical blood cultures. In addition to routine-collected hospital blood cultures, one parallel aerobic blood culture was collected and immediately refrigerated until being transported for ETGA analysis. After refrigeration holding and transport, parallel-collected cultures were placed into a BACTEC incubator and ETGA time-course analysis was performed. Of the 308 clinical blood cultures received, 22 were BACTEC positive, and thus were initially selected for ETGA time course analysis. The ETGA assay detected microbial growth in all 22 parallel-positive blood cultures in less time than a BACTEC incubator and also yielded genomic DNA for qPCR-based organism identification. In summary, feasibility of detecting microbes from clinical blood culture samples using the ETGA blood culture assay was demonstrated. Additional studies are being considered towards development of clinically beneficial versions of this methodology. PMID:24155986

  7. Streptokinase Clot Culture Compared with Whole Blood Culture for Isolation of Salmonella typhi and S. Paratyphi A from Patients with Enteric Fever

    DTIC Science & Technology

    1988-01-01

    Bravo, N. (1934). Diagnosis of typhoid fever using showed that the mnl STKCC was 28% more sensitive a string capsule device. Trsansma of da Rwal...masow and rose-spot whether Varidase, which is no longer available, would cultures for recovery nfSafialie &^ in typhoid fever . have been superior to...rate culture superior to sireptokinase clot cul:d8m 10 blood-to-broth ratio blood culture Achaowledgmenwat for diagnosia of typhoid fever . Annie Jenal

  8. Beacon-based (bbFISH®) technology for rapid pathogens identification in blood cultures.

    PubMed

    Sakarikou, Christina; Parisato, Martina; Lo Cascio, Giuliana; Fontana, Carla

    2014-04-22

    Diagnosis and treatment of bloodstream infections (BSI) are often hampered by the delay in obtaining the final results of blood cultures. Rapid identification of pathogens involved in BSI is of great importance in order to improve survival of septic patients. Beacon-based fluorescent in situ hybridization (hemoFISH® Gram positive and hemoFISH® Gram negative test kits, miacom diagnostics GmbH Düsseldorf, Germany) accelerates the identification of most frequent bacterial pathogens of sepsis. In this study a total of 558 blood culture (377 blood culture positive and 181 negative) were tested with the hemoFISH® method and the results were evaluated in comparison with the traditional culture based methods. The overall sensitivity and specificity of the hemoFISH® tests were 94.16% and 100%, while, the PPV and NPV were 100 and 89.16%, respectively. As the hemoFISH® results were obtained within 45 mins, the time difference between the final results of the traditional culture method and the hemoFISH® assay was about two days. Considering the good sensitivity and specificity of the hemoFISH® assays as well as the significant time saving in obtaining the final results (p-value 0.0001), the introduction of the system could be rialable in the microbiology laboratories, even alongside the traditional systems.

  9. Detection of parasitemia profiles by blood culture after treatment of human chronic Trypanosoma cruzi infection.

    PubMed

    de Castro, Ana Maria; Luquetti, Alejandro Ostermayer; Rassi, Anis; Chiari, Egler; Galvão, Lúcia Maria da Cunha

    2006-09-01

    The change in parasitemia profile, measured by sequential blood cultures of 27 benznidazole (Bz) treated patients compared with 13 untreated patients, on the chronic phase of chagas disease, is described. All patients were adults (age limits: 23-88) with positive serology (three tests); 23 of them were females. All patients were submitted to six blood cultures, three before and three after Bz treatment. The parasitemia was classified as nondetected (with three negative blood cultures), medium (one positive culture in three), and high (two or three positive cultures). From the eight patients with nondetected parasitemia before Bz, seven still had the same profile and only one switched to medium; from eight with medium parasitemia, seven shifted to nondetected, and one to high parasitemia. From the 11 patients with high parasitemia before Bz, ten converted to nondetected and only one was positive after Bz. Nineteen of the 27 patients changed the parasitemia profile (70.4%), and the rate of therapeutic failure was 11.1% (3/27) during the first 24 months of follow-up after Bz. The shift to nondetected parasitemia profile was from 8/27 to 24/27 patients after Bz treatment for the first 2 years. Only 46.2% (6/13) of the nontreated individuals changed the parasitemia profile. We conclude that there is a strong trypanocide effect of Bz (88.8%) and a rate of therapeutic failure of 11.1% during the first 2 years after trypanocidal treatment.

  10. Evaluating the culture of fetal erythroblasts from maternal blood for non-invasive prenatal diagnosis.

    PubMed

    Chen, H; Griffin, D K; Jestice, K; Hackett, G; Cooper, J; Ferguson-Smith, M A

    1998-09-01

    Fetal erythroblasts circulating in maternal blood are important candidate cells for non-invasive prenatal diagnosis. We have cultured erythroblasts from 16 maternal blood samples, both with and without prior enrichment by magnetic activated cell sorting (MACS), in a semi-solid medium containing growth factors. Individual colonies were examined by PCR with sex chromosome-specific primers and microsatellite marker primers. No conclusive Y-chromosome specific amplification could be demonstrated in any of the 16 cases, even when the mother was confirmed to be carrying a male fetus. All colonies tested by microsatellite marker PCR were of maternal origin. Our results suggest that the probability of obtaining fetal colonies from fetal erythroblasts circulating in maternal blood is very low and that approaches for culturing fetal erythroblasts in vitro cannot yet be used reliably for prenatal diagnosis using current methods for fetal cell enrichment.

  11. Improved diagnosis of orthopedic implant-associated infection by inoculation of sonication fluid into blood culture bottles.

    PubMed

    Portillo, María Eugenia; Salvadó, Margarita; Trampuz, Andrej; Siverio, Ana; Alier, Albert; Sorli, Lluisa; Martínez, Santos; Pérez-Prieto, Daniel; Horcajada, Juan P; Puig-Verdie, Lluis

    2015-05-01

    Sonication improved the diagnosis of orthopedic implant-associated infections (OIAI). We investigated the diagnostic performance of sonication fluid inoculated into blood culture bottles in comparison with that of intraoperative tissue and sonication fluid cultures. Consecutive patients with removed orthopedic hardware were prospectively included and classified as having OIAI or aseptic failure (AF) according to standardized criteria. The diagnostic procedure included the collection of five intraoperative tissue cultures and sonication of the removed device, followed by conventional culture of the sonication fluid. Cultures were incubated for 7 days (aerobic) or 14 days (anaerobic). In addition, 10 ml of sonication fluid was inoculated into each aerobic and anaerobic BacT/Alert FAN blood culture bottle and incubated in the automated blood culture system for 5 days. Of 75 included patients, 39 had OIAI and 36 AF. The sensitivity of sonication fluid inoculated into blood culture bottles (100%) was higher than that of conventional sonication fluid (87%; P = 0.05) or intraoperative tissue cultures (59%; P < 0.01). Previous antibiotic therapy reduced the culture sensitivity of conventional sonication fluid to 77% and that of intraoperative tissue to 55%, while it remained 100% for blood culture-inoculated sonication fluid. The time to positivity was shorter in blood culture-inoculated sonication fluid, with detection of 72% of microorganisms after 1 day of incubation, than for intraoperative tissue and conventional sonication fluid cultures, with detection of 18% and 28% of microorganisms, respectively. In conclusion, compared to conventional sonication fluid and intraoperative tissue cultures, sonication fluid inoculated into blood culture bottles improved the diagnosis of OIAI and considerably reduced the time to culture positivity.

  12. Evaluation of the FilmArray Blood Culture Identification Panel: Results of a Multicenter Controlled Trial.

    PubMed

    Salimnia, Hossein; Fairfax, Marilynn R; Lephart, Paul R; Schreckenberger, Paul; DesJarlais, Sharon M; Johnson, J Kristie; Robinson, Gwen; Carroll, Karen C; Greer, Amy; Morgan, Margie; Chan, Raymond; Loeffelholz, Michael; Valencia-Shelton, Frances; Jenkins, Stephen; Schuetz, Audrey N; Daly, Judy A; Barney, Trenda; Hemmert, Andrew; Kanack, Kristen J

    2016-03-01

    Sepsis is a major cause of morbidity, mortality, and increased medical expense. Rapid diagnosis improves outcomes and reduces costs. The FilmArray blood culture identification panel (BioFire Diagnostics LLC, Salt Lake City, UT), a highly multiplexed PCR assay, can identify 24 etiologic agents of sepsis (8 Gram-positive, 11 Gram-negative, and 5 yeast species) and three antimicrobial resistance genes (mecA, vanA/B, and blaKPC) from positive blood culture bottles. It provides results in about 1 h with 2 min for assay setup. We present the results of an eight-center trial comparing the sensitivity and specificity of the panel with those of the laboratories' standard phenotypic identification techniques, as well as with molecular methods used to distinguish Acinetobacter baumannii from other members of the A. calcoaceticus-A. baumannii complex and to detect antimicrobial resistance genes. Testing included 2,207 positive aerobic blood culture samples, 1,568 clinical and 639 seeded. Samples were tested fresh or were frozen for later testing within 8 h after the bottles were flagged as positive by an automated blood culture system. At least one organism was detected by the panel in 1,382 (88.1%) of the positive clinical specimens. The others contained primarily off-panel organisms. The panel reported multiple organisms in 81 (5.86%) positive clinical specimens. The unresolved blood culture identification sensitivity for all target detections exceeded 96%, except for Klebsiella oxytoca (92.2%), which achieved 98.3% sensitivity after resolution of an unavoidable phenotypic error. The sensitivity and specificity for vanA/B and blaKPC were 100%; those for mecA were 98.4 and 98.3%, respectively. Copyright © 2016 Salimnia et al.

  13. Evaluation of the FilmArray Blood Culture Identification Panel: Results of a Multicenter Controlled Trial

    PubMed Central

    Salimnia, Hossein; Lephart, Paul R.; Schreckenberger, Paul; DesJarlais, Sharon M.; Johnson, J. Kristie; Robinson, Gwen; Carroll, Karen C.; Greer, Amy; Morgan, Margie; Chan, Raymond; Loeffelholz, Michael; Valencia-Shelton, Frances; Jenkins, Stephen; Schuetz, Audrey N.; Daly, Judy A.; Barney, Trenda; Hemmert, Andrew; Kanack, Kristen J.

    2016-01-01

    Sepsis is a major cause of morbidity, mortality, and increased medical expense. Rapid diagnosis improves outcomes and reduces costs. The FilmArray blood culture identification panel (BioFire Diagnostics LLC, Salt Lake City, UT), a highly multiplexed PCR assay, can identify 24 etiologic agents of sepsis (8 Gram-positive, 11 Gram-negative, and 5 yeast species) and three antimicrobial resistance genes (mecA, vanA/B, and blaKPC) from positive blood culture bottles. It provides results in about 1 h with 2 min for assay setup. We present the results of an eight-center trial comparing the sensitivity and specificity of the panel with those of the laboratories' standard phenotypic identification techniques, as well as with molecular methods used to distinguish Acinetobacter baumannii from other members of the A. calcoaceticus-A. baumannii complex and to detect antimicrobial resistance genes. Testing included 2,207 positive aerobic blood culture samples, 1,568 clinical and 639 seeded. Samples were tested fresh or were frozen for later testing within 8 h after the bottles were flagged as positive by an automated blood culture system. At least one organism was detected by the panel in 1,382 (88.1%) of the positive clinical specimens. The others contained primarily off-panel organisms. The panel reported multiple organisms in 81 (5.86%) positive clinical specimens. The unresolved blood culture identification sensitivity for all target detections exceeded 96%, except for Klebsiella oxytoca (92.2%), which achieved 98.3% sensitivity after resolution of an unavoidable phenotypic error. The sensitivity and specificity for vanA/B and blaKPC were 100%; those for mecA were 98.4 and 98.3%, respectively. PMID:26739158

  14. Preparation of a blood culture pellet for rapid bacterial identification and antibiotic susceptibility testing.

    PubMed

    Croxatto, Antony; Prod'hom, Guy; Durussel, Christian; Greub, Gilbert

    2014-10-15

    Bloodstream infections and sepsis are a major cause of morbidity and mortality. The successful outcome of patients suffering from bacteremia depends on a rapid identification of the infectious agent to guide optimal antibiotic treatment. The analysis of Gram stains from positive blood culture can be rapidly conducted and already significantly impact the antibiotic regimen. However, the accurate identification of the infectious agent is still required to establish the optimal targeted treatment. We present here a simple and fast bacterial pellet preparation from a positive blood culture that can be used as a sample for several essential downstream applications such as identification by MALDI-TOF MS, antibiotic susceptibility testing (AST) by disc diffusion assay or automated AST systems and by automated PCR-based diagnostic testing. The performance of these different identification and AST systems applied directly on the blood culture bacterial pellets is very similar to the performance normally obtained from isolated colonies grown on agar plates. Compared to conventional approaches, the rapid acquisition of a bacterial pellet significantly reduces the time to report both identification and AST. Thus, following blood culture positivity, identification by MALDI-TOF can be reported within less than 1 hr whereas results of AST by automated AST systems or disc diffusion assays within 8 to 18 hr, respectively. Similarly, the results of a rapid PCR-based assay can be communicated to the clinicians less than 2 hr following the report of a bacteremia. Together, these results demonstrate that the rapid preparation of a blood culture bacterial pellet has a significant impact on the identification and AST turnaround time and thus on the successful outcome of patients suffering from bloodstream infections.

  15. Preparation of a Blood Culture Pellet for Rapid Bacterial Identification and Antibiotic Susceptibility Testing

    PubMed Central

    Croxatto, Antony; Prod'hom, Guy; Durussel, Christian; Greub, Gilbert

    2014-01-01

    Bloodstream infections and sepsis are a major cause of morbidity and mortality. The successful outcome of patients suffering from bacteremia depends on a rapid identification of the infectious agent to guide optimal antibiotic treatment. The analysis of Gram stains from positive blood culture can be rapidly conducted and already significantly impact the antibiotic regimen. However, the accurate identification of the infectious agent is still required to establish the optimal targeted treatment. We present here a simple and fast bacterial pellet preparation from a positive blood culture that can be used as a sample for several essential downstream applications such as identification by MALDI-TOF MS, antibiotic susceptibility testing (AST) by disc diffusion assay or automated AST systems and by automated PCR-based diagnostic testing. The performance of these different identification and AST systems applied directly on the blood culture bacterial pellets is very similar to the performance normally obtained from isolated colonies grown on agar plates. Compared to conventional approaches, the rapid acquisition of a bacterial pellet significantly reduces the time to report both identification and AST. Thus, following blood culture positivity, identification by MALDI-TOF can be reported within less than 1 hr whereas results of AST by automated AST systems or disc diffusion assays within 8 to 18 hr, respectively. Similarly, the results of a rapid PCR-based assay can be communicated to the clinicians less than 2 hr following the report of a bacteremia. Together, these results demonstrate that the rapid preparation of a blood culture bacterial pellet has a significant impact on the identification and AST turnaround time and thus on the successful outcome of patients suffering from bloodstream infections. PMID:25350577

  16. Evaluation of blood cultures in a children’s hospital located in Southeastern Anatolia

    PubMed Central

    Yiş, Reyhan

    2015-01-01

    Aim: Bloodstream infections in hospitalized patients are one of the most important causes of morbidity and mortality despite antimicrobial therapy. Early diagnosis and treatment of these infections is crucial. The aim of this study was to evaluate the distribution and antibiotic susceptibility of bacteria isolated from blood cultures in a children’s hospital in the Southeastern Anatolia during an 18-month period. Material and Methods: 7 040 blood cultures which were sent from hospitalized patients in Gaziantep Children’s Hospital between 01.07.2010 and 01.01.2012 were evaluated. Results: A total of 7 040 blood cultures were evaluated in this study. Microbial growth was detected in 2075 (29.47%) blood cultures. The most frequently isolated bacteria were coagulase-negative staphylococci (%45.97) which were followed by Salmonella spp. (%7.8). 12.12% of enterococcal isolates were resistant to glycopeptide antibiotics. The most frequently isolated gram negative bacterium was Salmonella spp. 15.43% of Salmonella spp. showed decreased susceptibility against quinolones. The ESBL positivity rate of E. coli and K. pneumoniae strains was found to be 35.08% and 57.14%, respectively. The imipenem resistance rate of P. aeruginosa was found to be 33.33%. The most common nonfermentative bacterium was S. maltophilia. Conclusions: The distribution of bacteria isolated from blood cultures and antibiotic resistance rates differ among different regions of Turkey. Different results obtained in our study may be related with regional tendencies to infections and patient population. Distribution of infectious agents and antibiotic resistance rates should be evaluated at regular intervals. This will lead to establishment of proper antibiotic usage policies in our country. PMID:26265894

  17. Evaluation of the nanosphere verigene gram-positive blood culture assay with the VersaTREK blood culture system and assessment of possible impact on selected patients.

    PubMed

    Beal, Stacy G; Ciurca, Jane; Smith, Geremy; John, Jeffrey; Lee, Francesca; Doern, Christopher D; Gander, Rita M

    2013-12-01

    The Verigene Gram-positive blood culture (BC-GP) assay (Nanosphere, Northbrook, IL) is a molecular method for the rapid identification of Gram-positive organisms and resistance markers directly from blood culture bottles. A total of 148 VersaTREK REDOX 1 40-ml aerobic bottles demonstrating Gram-positive bacteria were tested. Results were compared with those from conventional biochemical and matrix-assisted laser desorption ionization-time of flight (MALDI-TOF) identifications. We obtained isolates of methicillin-resistant Staphylococcus aureus (MRSA) (24), methicillin-susceptible Staphylococcus aureus (MSSA) (14), methicillin-resistant Staphylococcus epidermidis (MRSE) (17), methicillin-susceptible Staphylococcus epidermidis (MSSE) (9), other coagulase-negative staphylococci (19), Streptococcus salivarius (5), Streptococcus parasanguinis (2), Streptococcus sanguinis (1), Streptococcus cristatus (1), the Streptococcus bovis group (5), Streptococcus agalactiae (9), the Streptococcus anginosus group (1), Streptococcus pneumoniae (6), vancomycin-resistant Enterococcus faecium (VRE FCM) (16), vancomycin-susceptible Enterococcus faecalis (3), Aerococcus viridans (2), Bacillus (6), Corynebacterium (8), Lactobacillus (2), Micrococcus (2), Neisseria mucosa (1), Escherichia coli (3), Candida tropicalis (1), Propionibacterium (1), and Rothia (1). Overall agreement with the culture results was 95%. A total of 137 of 138 (99%) monomicrobial cultures were concordant. We tested 9 polymicrobial samples and found 33% agreement. A chart review of 31 patients with MRSA, MSSA, or VRE demonstrated that the Nanosphere BC-GP assay might have led to more appropriate antibiotic selection for these patients an average of 42 h earlier. Additionally, contact isolation could have been initiated an average of 37 h earlier for patients with MRSA or VRE. The BC-GP assay may have a positive impact on patient care, health care costs, and antibiotic stewardship.

  18. Molecular Detection of Streptococcus pneumoniae on Dried Blood Spots from Febrile Nigerian Children Compared to Culture.

    PubMed

    Iroh Tam, Pui-Ying; Hernandez-Alvarado, Nelmary; Schleiss, Mark R; Hassan-Hanga, Fatimah; Onuchukwu, Chuma; Umoru, Dominic; Obaro, Stephen K

    2016-01-01

    Nigeria has one of the highest burdens of pneumococcal disease in the world, but accurate surveillance is lacking. Molecular detection of infectious pathogens in dried blood spots (DBS) is an ideal method for surveillance of infections in resource-limited settings because of its low cost, minimal blood volumes involved, and ease of storage at ambient temperature. Our study aim was to evaluate a Streptococcus pneumoniae real-time polymerase chain reaction (rt-PCR) assay on DBS from febrile Nigerian children on Whatman 903 and FTA filter papers, compared to the gold standard of culture. Between September 2011 to May 2015, blood was collected from children 5 years of age or under who presented to six hospital study sites throughout northern and central Nigeria with febrile illness, and inoculated into blood culture bottles or spotted onto Whatman 903 or FTA filter paper. Culture and rt-PCR were performed on all samples. A total of 537 DBS specimens from 535 children were included in the study, of which 15 were culture-positive for S. pneumoniae. The rt-PCR assay detected S. pneumoniae in 12 DBS specimens (2.2%). One positive rt-PCR result was identified in a culture-negative specimen from a high-risk subject, and two positive rt-PCR results were negative on repeat testing. Six culture-confirmed cases of S. pneumoniae bacteremia were missed. Compared to culture, the overall sensitivities of Whatman 903 and FTA DBS for detection of S. pneumoniae were 57.1% (95% CI 18.4-90.1%) and 62.5% (95% CI 24.5-91.5%), respectively. Nonspecific amplification was noted in an additional 22 DBS (4.1%). Among these, six were positive for a non-S. pneumoniae pathogen on culture. Rt-PCR was able to detect S. pneumoniae from clinical DBS specimens, including from a culture-negative specimen. Our findings show promise of this approach as a surveillance diagnostic, but also raise important cautionary questions. Several DBS specimens were detected as S. pneumoniae by rt-PCR despite growth of

  19. Molecular Detection of Streptococcus pneumoniae on Dried Blood Spots from Febrile Nigerian Children Compared to Culture

    PubMed Central

    Iroh Tam, Pui-Ying; Hernandez-Alvarado, Nelmary; Schleiss, Mark R.; Hassan-Hanga, Fatimah; Onuchukwu, Chuma; Umoru, Dominic; Obaro, Stephen K.

    2016-01-01

    Background Nigeria has one of the highest burdens of pneumococcal disease in the world, but accurate surveillance is lacking. Molecular detection of infectious pathogens in dried blood spots (DBS) is an ideal method for surveillance of infections in resource-limited settings because of its low cost, minimal blood volumes involved, and ease of storage at ambient temperature. Our study aim was to evaluate a Streptococcus pneumoniae real-time polymerase chain reaction (rt-PCR) assay on DBS from febrile Nigerian children on Whatman 903 and FTA filter papers, compared to the gold standard of culture. Methods Between September 2011 to May 2015, blood was collected from children 5 years of age or under who presented to six hospital study sites throughout northern and central Nigeria with febrile illness, and inoculated into blood culture bottles or spotted onto Whatman 903 or FTA filter paper. Culture and rt-PCR were performed on all samples. Results A total of 537 DBS specimens from 535 children were included in the study, of which 15 were culture-positive for S. pneumoniae. The rt-PCR assay detected S. pneumoniae in 12 DBS specimens (2.2%). One positive rt-PCR result was identified in a culture-negative specimen from a high-risk subject, and two positive rt-PCR results were negative on repeat testing. Six culture-confirmed cases of S. pneumoniae bacteremia were missed. Compared to culture, the overall sensitivities of Whatman 903 and FTA DBS for detection of S. pneumoniae were 57.1% (95% CI 18.4–90.1%) and 62.5% (95% CI 24.5–91.5%), respectively. Nonspecific amplification was noted in an additional 22 DBS (4.1%). Among these, six were positive for a non-S. pneumoniae pathogen on culture. Conclusions Rt-PCR was able to detect S. pneumoniae from clinical DBS specimens, including from a culture-negative specimen. Our findings show promise of this approach as a surveillance diagnostic, but also raise important cautionary questions. Several DBS specimens were detected as

  20. Tobacco advertising in retail stores.

    PubMed

    Cummings, K M; Sciandra, R; Lawrence, J

    1991-01-01

    Recent studies have described tobacco advertising in the print media, on billboards, and through sponsorship of cultural and sporting events. However, little attention has been given to another common and unavoidable source of tobacco advertising, that which is encountered in retail stores. In July 1987, we conducted a survey of 61 packaged goods retail stores in Buffalo, NY, to assess the prevalence and type of point-of-sale tobacco advertising. In addition, store owners or managers were surveyed to determine their store's policy regarding tobacco advertising, receipt of monetary incentives from distributors for displaying tobacco ads, and willingness to display antitobacco ads. Six types of stores were involved in the study: 10 supermarkets, 10 privately owned grocery stores, 9 chain convenience food stores that do not sell gasoline, 11 chain convenience food stores that sell gasoline, 11 chain pharmacies, and 10 private pharmacies. Two-thirds of the stores displayed tobacco posters, and 87 percent had promotional items advertising tobacco products, primarily cigarettes. Larger stores, and those that were privately owned, tended to display more posters and promotional items. Eighty percent of tobacco product displays were for cigarettes, 16 percent for smokeless tobacco products, and 4 percent for cigars and pipe tobacco. Convenience stores selling gasoline had the most separate tobacco product displays. Of tobacco product displays, 24 percent were located adjacent to candy and snack displays. Twenty-nine of the 61 store owners or managers indicated that their store had a policy regulating the display of tobacco ads and tobacco product displays. Policies dealt primarily with the location of tobacco posters (for example, no ads in the window) and number of product displays. Only 14 shop owners or managers indicated that they had previously displayed antitobacco information; more than half (31 of 61) said that they would be willing to display antitobaccoads.In many

  1. [Identification of staphylococci directly from positive blood culture bottles by MALDI-TOF MS system].

    PubMed

    Kilic, Abdullah; Baysallar, Mehmet

    2014-07-01

    Bloodstream infections are substantial causes of morbidity and mortality worldwide. Staphylococcus species are the most commonly isolated microorganisms from blood cultures in clinical microbiology laboratories. MALDI-TOF MS (Matrix-Assisted Laser Desorption/Ionization Time- of- Flight Mass Spectrometry) system allows the identification of microorganisms directly from positive blood culture bottles. The aim of this study was to evaluate the MALDI-TOF MS system for the identification of staphylococci directly from the positive blood culture bottles which revealed the presence of gram-positive cocci by staining. A total of 96 positive blood culture bottles that yielded gram-positive cocci by Gram stain were evaluated. These blood cultures were obtained from 69 patients between December 2013-February 2014. Conventional methods and BD Phoenix™ automated bacterial identification system (Becton Dickinson, USA) were used for routine identification. The strains were also identified by real-time Taqman PCR (qPCR) which was considered as the reference method. In MALDI-TOF MS method, MALDI Sepsityper™ Kit was used for the bacterial identification and all measurements were carried out by using Microflex LT instrument and FlexControl 3.0 software (Bruker Daltonics, USA). Of 96 culture bottles positive for gram-positive cocci, 90 were correctly identified as staphylococci at genus level with all the three study methods (qPCR, BD Phoenix, Bruker MALDI-TOF MS). The other six samples were identified as Enterococcus faecium (n= 4) and Streptococcus pyogenes (n= 2) by both Phoenix and the MALDI-TOF systems. Of the 90 samples, 87 were identified at the species level (15 S.aureus, 33 S.epidermidis, 29 S.hominis, 10 S.haemolyticus) and three at the genus level by the reference qPCR method. When comparing the results obtained by qPCR and Bruker MALDI-TOF MS, incompatibility was detected for three isolates. Those isolates were identified as S.hominis by qPCR, however two of them were

  2. Effects of transforming growth factor-beta on long-term human cord blood monocyte cultures

    SciTech Connect

    Orcel, P.; Bielakoff, J.; De Vernejoul, M.C. )

    1990-02-01

    Transforming growth factor-beta (TGF-beta) modulates growth and differentiation in many cell types and is abundant in bone matrix. We recently showed that human cord blood monocytes cultured in the presence of 1,25(OH)2D3 acquire some features of osteoclast precursors. Since TGF-beta has been shown to influence bone resorption in organ culture, we have studied the effect of TGF-beta (1-1,000 pg/ml) on cord blood monocyte cultures. These cells were cultured on plastic substrate during 3 weeks in the presence of 20% horse serum and 10(-9) M 1,25(OH)2D3. TGF-beta, from a concentration of 10 pg/ml in the culture medium, decreased in a dose dependent manner the formation of multinucleated cells. At a concentration of TGF-beta of 1 ng/ml, the multinucleated cells were reduced to 2.1% +/- 0.3%, compared to 19.3% +/- 1.5% in control cultures. TGF-beta inhibited in a dose-dependent manner the proliferation of cord blood monocytes as assessed by 3H-thymidine incorporation at 7 and 14 days of culture. The fusion index was also decreased by 3 weeks of treatment with TGF-beta. Indomethacin did not reverse the inhibitory effects of TGF-beta. The expression of the osteoclastic phenotype was assessed using two different antibodies: 23C6, a monoclonal antibody directed against the vitronectin receptor, which is highly expressed by osteoclasts but not by adult monocytes, and an antibody to HLA-DR, which is not present on osteoclast. TGF-beta decreased the expression of HLA-DR and increased in a dose-dependent manner the proportion of 23C6-labeled cells; these results suggest that TGF-beta could modulate a differentiation effect to the osteoclastic phenotype. However, when cord blood monocytes were cultured on devitalized rat calvariae prelabeled with 45Ca, TGF-beta did not induce any 45Ca release from bone cultured with monocytes.

  3. Direct Antimicrobial Susceptibility Testing of Gram-Negative Bacilli in Blood Cultures by an Electrochemical Method

    PubMed Central

    Huang, Ay Huey; Wu, Jiunn Jong; Weng, Yu Mei; Ding, Hwia Cheng; Chang, Tsung Chain

    1998-01-01

    Nonfastidious aerobic gram-negative bacilli (GNB) are commonly isolated from blood cultures. The feasibility of using an electrochemical method for direct antimicrobial susceptibility testing of GNB in positive blood cultures was evaluated. An aliquot (10 μl) of 1:10-diluted positive blood cultures containing GNB was inoculated into the Bactometer module well (bioMérieux Vitek, Hazelwood, Mo.) containing 1 ml of Mueller-Hinton broth supplemented with an antibiotic. Susceptibility tests were performed in a breakpoint broth dilution format, with the results being categorized as resistant, intermediate, or susceptible. Seven antibiotics (ampicillin, cephalothin, gentamicin, amikacin, cefamandole, cefotaxime, and ciprofloxacin) were used in this study, with each agent being tested at the two interpretive breakpoint concentrations. The inoculated modules were incubated at 35°C, and the change in impedance in each well was continuously monitored for 24 h by the Bactometer. The MICs of the seven antibiotics for each blood isolate were also determined by the standardized broth microdilution method. Of 146 positive blood cultures (1,022 microorganism-antibiotic combinations) containing GNB tested by the direct method, the rates of very major, major, and minor errors were 0, 1.1, and 2.5%, respectively. The impedance method was simple; no centrifugation, preincubation, or standardization of the inocula was required, and the susceptibility results were normally available within 3 to 6 h after inoculation. The rapid method may allow proper antimicrobial treatment almost 30 to 40 h before the results of the standard methods are available. PMID:9738038

  4. Optimal turnaround time for direct identification of microorganisms by mass spectrometry in blood culture.

    PubMed

    Randazzo, Adrien; Simon, Marc; Goffinet, Pierre; Classen, Jean-François; Hougardy, Nicolas; Pierre, Pascal; Kinzinger, Philippe; Mauel, Etienne; Goffinet, Jean-Sébastien

    2016-11-01

    During the past few years, several studies describing direct identification of bacteria from blood culture using mass spectrometry have been published. These methods cannot, however, be easily integrated into a common laboratory workflow because of the high hands-on time they require. In this paper, we propose a new method of identification with a short hands-on time and a turnaround time shorter than 15min. Positive blood bottles were homogenised and 600μL of blood were transferred to an Eppendorf tube where 600μL of lysis buffer were added. After homogenisation, a centrifugation step of 4min at 10,500g was performed and the supernatant was discarded. The pellet was then washed and loaded in quadruplicate into wells of a Vitek® MS-DS plate. Each well was covered with a saturated matrix solution and a MALDI-TOF mass spectrometry analysis was performed. Species were identified using the software Myla 3.2.0-2. We analysed 266 positive blood culture bottles. A microorganism grew in 261 cultures, while five bottles remained sterile after 48h of incubation in subculture. Our method reaches a probability of detection at the species level of 77.8% (203/261) with a positive predictive value of 99.5% (202/203). We developed a new method for the identification of microorganisms using mass spectrometry, directly performed from a positive blood culture. This method has short hands-on time and turnaround time and can easily take place in the workflow of a laboratory, with comparable results in performance with other methods reported in the literature. Copyright © 2016 Elsevier B.V. All rights reserved.

  5. A rapid, highly sensitive and culture-free detection of pathogens from blood by positive enrichment.

    PubMed

    Vutukuru, Manjula Ramya; Sharma, Divya Khandige; Ragavendar, M S; Schmolke, Susanne; Huang, Yiwei; Gumbrecht, Walter; Mitra, Nivedita

    2016-12-01

    Molecular diagnostics is a promising alternative to culture based methods for the detection of bloodstream infections, notably due to its overall lower turnaround time when starting directly from patient samples. Whole blood is usually the starting diagnostic sample in suspected bloodstream infections. The detection of low concentrations of pathogens in blood using a molecular assay necessitates a fairly high starting volume of blood sample in the range of 5-10mL. This large volume of blood sample has a substantial accompanying human genomic content that interferes with pathogen detection. In this study, we have established a workflow using magnetic beads coated with Apolipoprotein H that makes it possible to concentrate pathogens from a 5.0mL whole blood sample, thereby enriching pathogens from whole blood background and also reducing the sample volume to ~200μL or less. We have also demonstrated that this method of enrichment allows detection of 1CFU/mL of Escherichia coli, Enterococcus gallinarum and Candida tropicalis from 5mL blood using quantitative PCR; a detection limit that is not possible in unenriched samples. The enrichment method demonstrated here took 30min to complete and can be easily integrated with various downstream molecular and microbiological techniques. Copyright © 2016 Elsevier B.V. All rights reserved.

  6. Long-Term Stored Hemoglobin-Vesicles, a Cellular Type of Hemoglobin-Based Oxygen Carrier, Has Resuscitative Effects Comparable to That for Fresh Red Blood Cells in a Rat Model with Massive Hemorrhage without Post-Transfusion Lung Injury

    PubMed Central

    Yamasaki, Keishi; Sakai, Hiromi; Otagiri, Masaki

    2016-01-01

    Hemoglobin-vesicles (HbV), encapsulating highly concentrated human hemoglobin in liposomes, were developed as a substitute for red blood cells (RBC) and their safety and efficacy in transfusion therapy has been confirmed in previous studies. Although HbV suspensions are structurally and physicochemically stabile for least 1-year at room temperature, based on in vitro experiments, the issue of whether the use of long-term stored HbV after a massive hemorrhage can be effective in resuscitations without adverse, post-transfusion effects remains to be clarified. We report herein on a comparison of the systemic response and the induction of organ injuries in hemorrhagic shock model rats resuscitated using 1-year-stored HbV, freshly packed RBC (PRBC-0) and by 28-day-stored packed RBC (PRBC-28). The six-hour mortality after resuscitation was not significantly different among the groups. Arterial blood pressure and blood gas parameters revealed that, using HbV, recovery from the shock state was comparable to that when PRBC-0 was used. Although no significant change was observed in serum parameters reflecting liver and kidney injuries at 6 hours after resuscitation among the three resuscitation groups, results based on Evans Blue and protein leakage in bronchoalveolar lavage fluid, the lung wet/dry weight ratio and histopathological findings indicated that HbV as well as PRBC-0 was less predisposed to result in a post-transfusion lung injury than PRBC-28, as evidenced by low levels of myeloperoxidase accumulation and subsequent oxidative damage in the lung. The findings reported herein indicate that 1-year-stored HbV can effectively function as a resuscitative fluid without the induction of post-transfused lung injury and that it is comparable to fresh PRBC, suggesting that HbV is a promising RBC substitute with a long shelf-life. PMID:27798697

  7. Towards cultural materialism in the medical humanities: the case of blood rejuvenation.

    PubMed

    Oakley, Catherine

    2017-05-11

    This paper argues for an approach within the medical humanities that draws on the theoretical legacy of cultural materialism as a framework for reading cultural practices and their relationship to the social and economic order. It revisits the origins and development of cultural materialism in cultural studies and literary studies between the 1970s and 1990s and considers how, with adaptation, this methodology might facilitate ideological criticism focused on material formations of health, disease and the human body. I outline three key characteristics of a medicocultural materialist approach along these lines: (a) interdisciplinary work on a broad range of medical and cultural sources, including those drawn from 'popular' forms of culture; (b) the combination of historicist analysis with scrutiny of present-day contexts; (c) analyses that engage with political economy perspectives and/or the work of medical sociology in this area. The subsequent sections of the paper employ a medicocultural materialist approach to examine conjectural understandings of, and empirical investigations into, the capacity of transfused human blood to rejuvenate the ageing body. I trace textual faultlines that expose the structures of power which inform the movement of blood between bodies in 'medical gothic' fictions from the 19th-century fin de siècle, including Mary Elizabeth Braddon's 'Good Lady Ducayne' (1896) and Bram Stoker's Dracula (1897). I conclude with a critique of biomedical innovations in blood rejuvenation in the era of medical neoliberalism, before considering the potential applications of medicocultural materialism to other topics within the field of the medical humanities. Published by the BMJ Publishing Group Limited. For permission to use (where not already granted under a licence) please go to http://www.bmj.com/company/products-services/rights-and-licensing/.

  8. Comparison of acridine orange, methylene blue, and Gram stains for blood cultures.

    PubMed Central

    Mirrett, S; Lauer, B A; Miller, G A; Reller, L B

    1982-01-01

    Direct microscopic screening of blood cultures by Gram stain or methylene blue stain is time consuming and frequently insensitive. Therefore, we evaluated a fluorescent-staining procedure that uses acridine orange (AO) at pH 3.5 and compared it with the methylene blue and Gram stain procedures. All smears were prepared within 24 h of receiving the culture, fixed with methanol, and examined without the results of the companion smears being known. AO-stained smears were examined with incident-light fluorescence at 600 x magnification and confirmed at 1,500x magnification. All bottles macroscopically positive within 24 h were excluded from the study. Of 2,946 cultures entered into the study, 204 (6.9%) were positive within 3 days. The sensitivity and specificity of AO based on these culture results were 52 and 98%, respectively, compared with 38% sensitivity and 99% specificity by methylene blue and Gram stains. The AO staining procedure is a simple, sensitive, screening technique for the early detection of positive blood cultures. PMID:6175656

  9. Characterization of glucocerebrosidase in peripheral blood cells and cultured blastoid cells.

    PubMed

    Aerts, J M; Heikoop, J; van Weely, S; Donker-Koopman, W E; Barranger, J A; Tager, J M; Schram, A W

    1988-08-01

    We have characterized glucocerebrosidase in various cell types of peripheral blood of control subjects and in cultured human blastoid cells. The intracellular level of glucocerebrosidase in cultured blastoid cells (10-30 nmol substrate hydrolyzed/h.mg protein) resembles closely values observed for leukocyte cell types and various tissues and is significantly lower than that observed in cultured fibroblasts (150-500 nmol substrate hydrolyzed/h.mg protein). Glucocerebrosidase is extracted from leukocyte cell types and cultured blastoid cells almost exclusively in a monomeric, nonactivated form with enzymatic properties identical to those of the tissue enzyme. In contrast, extracts of platelets are rich in an aggregated, activated form of the enzyme. Glucocerebrosidase in blood cells and cultured blastoid cells is heterogeneous with respect to Mr and pI due to a heterogeneous oligosaccharide composition of the enzyme. The different forms seen represent intermediates in the biosynthesis and maturation of the enzyme. Blastoid cells should thus be an attractive model system for studying the natural history of glucocerebrosidase in a cell type related to those cells involved in the pathology of Gaucher disease.

  10. Continuous and semi-continuous cell culture for production of blood clotting factors.

    PubMed

    Desai, Sunil G

    2015-11-10

    Recombinant clotting factors are important biotherapeutics that Pfizer has produced and marketed for over fifteen years. Owing to the complexity of the structure and function of these blood factors, it can be challenging to achieve the required product quality and manufacturing productivity. The article highlights the semi-continuous and continuous cell culture processes employed by Pfizer for the production of BeneFIX and ReFacto AF. The benefits of such processes, the challenges of maintaining an aseptic production culture for extended periods, and batch definition are discussed in this article. Copyright © 2015 Elsevier B.V. All rights reserved.

  11. Rapid identification and antimicrobial susceptibility profiling of Gram-positive cocci in blood cultures with the Vitek 2 system.

    PubMed

    Lupetti, A; Barnini, S; Castagna, B; Capria, A-L; Nibbering, P H

    2010-01-01

    Rapid identification and antimicrobial susceptibility profiling of the bacteria in blood cultures can result in clinical and financial benefits. Addition of saponin to the fluid from blood culture bottles promotes the recovery of the bacteria and thus may shorten the turnaround time of the microbiological analyses. In this study we compared the identification and susceptibility profiles of saponin-treated and untreated (standard method) blood cultures monomicrobial for Gram-positive cocci using Vitek 2. We concordantly identified 49 (89%) of 55 monobacterial cultures using the results with the standard method as reference. Complete categorical agreement between the susceptibility profiles with the new and the standard method was found for 26 (53%) of 49 isolates, while discrepancies were seen for 23 (47%) cultures. E-tests indicated that the new method resulted in a correct susceptibility profile for 8 (35%) of these 23 blood cultures. Therefore, 34 (69%) of 49 cultures showed a concordant/correct susceptibility profile for all antimicrobials with an overall error rate of 2.3%. Thus, addition of saponin to the fluid from blood culture bottles of the Bactec 9240 leads to the rapid (results available >or=12 hours earlier) and reliable identification and susceptibility profiling of Gram-positive cocci in blood cultures with Vitek 2.

  12. Blood Culture Collection through Peripheral Intravenous Catheters Increases the Risk of Specimen Contamination among Adult Emergency Department Patients

    PubMed Central

    Self, Wesley H.; Speroff, Theodore; McNaughton, Candace D.; Wright, Patty W.; Miller, Geraldine; Johnson, James G.; Daniels, Titus L.; Talbot, Thomas R.

    2017-01-01

    Five hundred five blood cultures collected through a peripheral intravenous catheter (PIV) in an emergency department were matched to cultures obtained by dedicated venipuncture from the same patient within 10 minutes. The relative risk of contamination for cultures collected through PIVs compared with dedicated venipuncture was 1.83 (95% confidence interval, 1.08–3.11). PMID:22476282

  13. Blood culture collection through peripheral intravenous catheters increases the risk of specimen contamination among adult emergency department patients.

    PubMed

    Self, Wesley H; Speroff, Theodore; McNaughton, Candace D; Wright, Patty W; Miller, Geraldine; Johnson, James G; Daniels, Titus L; Talbot, Thomas R

    2012-05-01

    Five hundred five blood cultures collected through a peripheral intravenous catheter (PIV) in an emergency department were matched to cultures obtained by dedicated venipuncture from the same patient within 10 minutes. The relative risk of contamination for cultures collected through PIVs compared with dedicated venipuncture was 1.83 (95% confidence interval, 1.08-3.11).

  14. Direct blood culturing on solid medium outperforms an automated continuously monitored broth-based blood culture system in terms of time to identification and susceptibility testing

    PubMed Central

    Idelevich, E.A.; Grünastel, B.; Peters, G.; Becker, K.

    2015-01-01

    Pathogen identification and antimicrobial susceptibility testing (AST) should be available as soon as possible for patients with bloodstream infections. We investigated whether a lysis-centrifugation (LC) blood culture (BC) method, combined with matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS) identification and Vitek 2 AST, provides a time advantage in comparison with the currently used automated broth-based BC system. Seven bacterial reference strains were added each to 10 mL human blood in final concentrations of 100, 10 and 1 CFU/mL. Inoculated blood was added to the Isolator 10 tube and centrifuged at 3000 g for 30 min, then 1.5 mL sediment was distributed onto five 150-mm agar plates. Growth was observed hourly and microcolonies were subjected to MALDI-TOF MS and Vitek 2 as soon as possible. For comparison, seeded blood was introduced into an aerobic BC bottle and incubated in the BACTEC 9240 automated BC system. For all species/concentration combinations except one, successful identification and Vitek 2 inoculation were achieved even before growth detection by BACTEC. The fastest identification and inoculation for AST were achieved with Escherichia coli in concentrations of 100 CFU/mL and 10 CFU/mL (after 7 h each, while BACTEC flagged respective samples positive after 9.5 h and 10 h). Use of the LC-BC method allows skipping of incubation in automated BC systems and, used in combination with rapid diagnostics from microcolonies, provides a considerable advantage in time to result. This suggests that the usefulness of direct BC on solid medium should be re-evaluated in the era of rapid microbiology. PMID:26909155

  15. Direct blood culturing on solid medium outperforms an automated continuously monitored broth-based blood culture system in terms of time to identification and susceptibility testing.

    PubMed

    Idelevich, E A; Grünastel, B; Peters, G; Becker, K

    2016-03-01

    Pathogen identification and antimicrobial susceptibility testing (AST) should be available as soon as possible for patients with bloodstream infections. We investigated whether a lysis-centrifugation (LC) blood culture (BC) method, combined with matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS) identification and Vitek 2 AST, provides a time advantage in comparison with the currently used automated broth-based BC system. Seven bacterial reference strains were added each to 10 mL human blood in final concentrations of 100, 10 and 1 CFU/mL. Inoculated blood was added to the Isolator 10 tube and centrifuged at 3000 g for 30 min, then 1.5 mL sediment was distributed onto five 150-mm agar plates. Growth was observed hourly and microcolonies were subjected to MALDI-TOF MS and Vitek 2 as soon as possible. For comparison, seeded blood was introduced into an aerobic BC bottle and incubated in the BACTEC 9240 automated BC system. For all species/concentration combinations except one, successful identification and Vitek 2 inoculation were achieved even before growth detection by BACTEC. The fastest identification and inoculation for AST were achieved with Escherichia coli in concentrations of 100 CFU/mL and 10 CFU/mL (after 7 h each, while BACTEC flagged respective samples positive after 9.5 h and 10 h). Use of the LC-BC method allows skipping of incubation in automated BC systems and, used in combination with rapid diagnostics from microcolonies, provides a considerable advantage in time to result. This suggests that the usefulness of direct BC on solid medium should be re-evaluated in the era of rapid microbiology.

  16. Impact of reporting gram stain results from blood culture bottles on the selection of antimicrobial agents.

    PubMed

    Uehara, Yuki; Yagoshi, Michiko; Tanimichi, Yumiko; Yamada, Hiroko; Shimoguchi, Kazuo; Yamamoto, Sachiyo; Yanai, Mitsuru; Kumasaka, Kazunari

    2009-07-01

    We assessed the usefulness of reporting direct blood Gram stain results compared with the results of positive blood cultures in 482 episodes and monitored impact on selection of antimicrobial treatment. We found that the reporting groups "Staphylococcus spp," "Pseudomonas spp and related organisms," and "yeasts" identified in this way matched perfectly with later culture identification. When the report indicated Staphylococcus spp or Pseudomonas spp and related organisms, physicians started or changed antimicrobials suitable for these bacteria more frequently than when "other streptococci" and "family Enterobacteriaceae" were reported (P < .05). Incorrect recognition of Acinetobacter spp as Enterobacteriaceae family is still the most challenging problem in this context. Gram stain results that definitively identify Staphylococcus spp, Pseudomonas spp and related organisms, and yeasts reliably can be rapidly provided by clinical laboratories; this information has a significant impact on early selection of effective antimicrobials. Further investigation is needed to assess the clinical impact of reporting Gram stain results in bacteremia.

  17. Antimicrobial susceptibility profile of Acinetobacter species isolated from blood cultures in two Japanese university hospitals.

    PubMed

    Kishii, Kozue; Kikuchi, Ken; Yoshida, Atsushi; Okuzumi, Katsuko; Uetera, Yushi; Yasuhara, Hiroshi; Moriya, Kyoji

    2014-02-01

    Carbapenem-resistant Acinetobacter baumannii has rapidly spread worldwide. This study investigated antibiotic susceptibility and genotypic resistance of 123 consecutive blood culture isolates of Acinetobacter species collected between 2003 and 2011 in two Japanese hospitals. The isolates were assigned to 13 species. Carbapenem resistance was detected in four isolates. Only one A. baumannii isolate had blaOXA-23 together with ISAba1; the remaining three isolates had IMP-1 metallo-β-lactamase. Quinolone resistance was detected in five isolates that had point mutations in the quinolone resistance-determining region. The predominance of various non-A. baumannii species and low prevalence of carbapenem resistance among blood culture isolates of Acinetobacter species in two Japanese hospitals were confirmed.

  18. Evaluation of substrates for radiometric detection of bacteria in blood cultures

    SciTech Connect

    Bopp, H.; Ellner, P.D.

    1988-05-01

    Various /sup 14/C-labeled substrates were evaluated for their potential use in blood culture media. These uniformly labeled compounds were added to hypertonic and anaerobic formulations of modified Columbia broth and compared with analogous BACTEC media with the BACTEC 460. Different bacterial species gave significant growth indices when 2.0 microCi of labeled glucose, glutamic acid, aspartic acid, arginine, or formate was used alone or in combinations in the experimental media. The combination of glucose, glutamic acid, and sodium formate was selected, and simulated blood cultures with representative aerobic, facultative, and anaerobic bacteria and a yeast were compared with BACTEC vials. Under these conditions, the experimental media often became positive several hours earlier than the BACTEC vials and usually produced higher growth indices.

  19. Validation of a monoclonal antibody-based immunofluorescent assay to detect Burkholderia pseudomallei in blood cultures.

    PubMed

    Dulsuk, Adul; Paksanont, Suporn; Sangchankoom, Adisak; Ekchariyawat, Peeraya; Phunpang, Rungnapa; Jutrakul, Yaowaruk; Chantratita, Narisara; West, T Eoin

    2017-01-22

    Identification of Burkholderia pseudomallei, the cause of melioidosis, using routine methods takes several days. Use of a monoclonal antibody-based immunofluorescent assay (IFA) on positive blood cultures may speed diagnosis. We tested the diagnostic accuracy of the IFA on 545 blood cultures positive for Gram-negative organisms at Udon Thani Hospital, Thailand, between June 2015 and August 2016. Sensitivity of the IFA was 100% and specificity was 99.6%. The median decrease in time to pathogen identification between the IFA result and routine methods was 28 h (IQR 25-51), p<0.0001. The IFA accurately expedites the diagnosis of melioidosis. © The Author 2017. Published by Oxford University Press on behalf of Royal Society of Tropical Medicine and Hygiene.

  20. Bacterial and fungal DNA extraction from positive blood culture bottles: a manual and an automated protocol.

    PubMed

    Mäki, Minna

    2015-01-01

    When adapting a gene amplification-based method in a routine sepsis diagnostics using a blood culture sample as a specimen type, a prerequisite for a successful and sensitive downstream analysis is the efficient DNA extraction step. In recent years, a number of in-house and commercial DNA extraction solutions have become available. Careful evaluation in respect to cell wall disruption of various microbes and subsequent recovery of microbial DNA without putative gene amplification inhibitors should be conducted prior selecting the most feasible DNA extraction solution for the downstream analysis used. Since gene amplification technologies have been developed to be highly sensitive for a broad range of microbial species, it is also important to confirm that the used sample preparation reagents and materials are bioburden-free to avoid any risks for false-positive result reporting or interference of the diagnostic process. Here, one manual and one automated DNA extraction system feasible for blood culture samples are described.

  1. Clinical significance of Staphylococcus saprophyticus identified on blood culture in a tertiary care hospital.

    PubMed

    Choi, Sang-Ho; Woo, Jun Hee; Jeong, Jin-Yong; Kim, Nam Joong; Kim, Mi-Na; Kim, Yang Soo; Ryu, Jiso

    2006-11-01

    Staphylococcus saprophyticus is a well-known cause of acute uncomplicated urinary tract infection in young women. However, the clinical significance of this organism isolated from blood culture has not been determined. We assessed the clinical significance and characteristics of S. saprophyticus identified on blood culture. A total of 24 patients were identified, and 7 patients (29.2%) were considered to have clinically significant bacteremia. Of the 7 patients with clinically significant bacteremia, hematologic malignancy was the most common underlying illness (5 patients), and tunneled-central venous catheter was the most common portal of entry (4 patients). In no case did S. saprophyticus bacteremia originate from the urinary tract. One patient died during hospitalization. However, the death was not directly related to bacteremia. In conclusion, our data suggest that bacteremia caused by S. saprophyticus is most commonly associated with tunneled-central venous catheter in patients with hematologic malignancies and may be associated with a lower risk of mortality.

  2. [PCR rDNA 16S used for the etiological diagnosis of blood culture negative endocarditis].

    PubMed

    Baty, G; Lanotte, P; Hocqueloux, L; Prazuck, T; Bret, L; Romano, M; Mereghetti, L

    2010-06-01

    We report the case of a 55 year-old man presenting with a double aortic and mitral endocarditis for which resected valve culture was repeatedly negative. Specific PCR made on valves because of highly positive blood tests for Bartonella henselae remained negative. A molecular approach was made with 16S rDNA PCR, followed by sequencing. Bartonella quintana was identified as the etiology of endocarditis. B. quintana, "fastidious" bacteria, even if hard to identify in a laboratory, is often reported as a blood culture negative endocarditis (BCNE) agent. Molecular biology methods have strongly improved the diagnosis of BCNE. We propose a review of the literature focusing on the interest of broad-spectrum PCR on valve for the etiological diagnosis of BCNE.

  3. Blood Transfusion and Donation

    MedlinePlus

    ... receiving the blood transfusion. To keep blood safe, blood banks carefully screen donated blood. The risk of catching ... one or more times before the surgery. A blood bank will store your blood for your use. NIH: ...

  4. Extensive proliferative capacity of single isolated CD34 human cord blood cells in suspension culture.

    PubMed

    Xiao, M; Broxmeyer, H E; Horie, M; Grigsby, S; Lu, L

    1994-01-01

    Nonadherent, low-density T-lymphocyte-depleted (NALT-) CD34 cells from normal human cord blood were assessed in suspension culture for the effects of recombinant cytokines on their proliferation, differentiation, and generation of myeloid progenitor cells. In this cell population, 82% of cells expressed c-kit protein as assessed by in situ hybridization, and their cloning efficiency was 85% when cells were plated at low cell numbers with combinations of growth factors. CD34 cells were sorted as 1, 5, or 10 cell(s) per well and also at 5000 cells per dish to initiate stromal-free suspension cultures in the presence of steel factor (SLF), interleukin (IL)-1 alpha, and IL-3. Forty-eight percent of the wells started with a single CD34 cell were positive for growth after 14 days, and the wells contained greater than 5 x 10(3) cells by 21-28 days. Progenitors were assayed weekly with cultures initiated with 1 or 5000 cells. While the fold expansion of nucleated cells was greater in cultures initiated with one cell per well (> 5000 compared to 791-fold expansion for 5000 cells), the fold expansion of progenitors was greater than 5000 cells were used to initiate cultures. Under optimal conditions, there was, respectively, a 160-, 164-, and 57-fold output of high proliferative potential colony-forming cells, granulocyte-macrophage colony-forming units, and erythroid burst-forming units/granulocyte erythroid macrophage megakaryocyte colony-forming units within 1-3 weeks for cultures initiated with 5000 CD34 cells compared with respective fold increases of 29, 16, and 1, for single-initiated cultures. These results demonstrate the expansion capacity of single CD34 cord blood cells and demonstrate that factors in addition to SLF, IL-1 alpha, and IL-3 are necessary for optimal expansion of progenitors from single isolated CD34 cells.

  5. [Utility of prolonged incubation and terminal subcultures of blood cultures from immunocompromised patients].

    PubMed

    Soloaga, R; Procopio, A; Manganello, S; Ivanovic, V; Romay, N; Pirosanto, Y; Fernández, A; Zudiker, R; Echeverría, A; Nagel, C; del Castillo, M; López, E; Gutfraind, Z; Tokumoto, M; Guelfand, L

    2001-01-01

    The value of blind terminal subcultures (7 and 30 days) and prolonged incubation (30 days) of blood cultures from immunosuppressed patients was analyzed in the Fundación Favaloro, the Fundación para la Lucha contra las Enfermedades Neurológicas de la Infancia and the Hospital de Niños Ricardo Gutiérrez. A total of 2707 blood cultures and 369 patients were included (transplantation of solid organs 154, oncohematologic disorders 106 and solid tumors 109). Bact-Alert bottles were incubated at 35 degrees C for 30 days in the Bact-Alert System. Bottles with positive signals were routinely removed, and aliquots of the broth were Gram stained and subcultured aerobically in chocolate agar and Sabouraud agar. A total of 136 bacteremic episodes were obtained. The positivization time of blood cultures was 81.6% at 24 h, 93.3% at 48 h, 94.5% at 72 h and 97.7% within 7 days. Only 3 (2.2%) episodes were positive by blind terminal subcultures and 1 (0.75%) by prolonged incubation (14 days). The median time and range of positivization in hours were 13.8 and 2.2-168, respectively. The microorganisms isolated were coagulase negative staphylococci (n = 24), Klebsiella pneumoniae (n = 22), Staphylococcus aureus (n = 21), Escherichia coli (n = 18), Acinetobacter spp (n = 9), Candida spp (n = 8), Pseudomonas aeruginosa (n = 6), Enterobacter cloacae (n = 5), Stenotrophomonas maltophilia (n = 5), Enterococcus faecalis, Salmonella spp and Capnocytophaga sputigena (n = 2), Enterobacter aerogenes, Enterococcus faecium, Citrobacter diversus, Candida albicans, Klebsiella oxytoca, Chryseomonas luteola, Serratia marcescens, Abiotrophia spp, Campylobacter jejuni, Moraxella catarrhalis, Moraxella urethralis, Neisseria sicca, beta hemolytic group G streptococci, Rhodococcus equi, Micrococcus spp, Cryptococcus neoformans and Streptococcus mitis (n = 1). In our experience, blind terminal subcultures and prolonged incubation of blood cultures from immunosuppressed patients are unnecessary and

  6. Blood culture contamination with Enterococci and skin organisms: implications for surveillance definitions of primary bloodstream infections.

    PubMed

    Freeman, Joshua T; Chen, Luke Francis; Sexton, Daniel J; Anderson, Deverick J

    2011-06-01

    Enterococci are a common cause of bacteremia but are also common contaminants. In our institution, approximately 17% of positive blood cultures with enterococci are mixed with skin organisms. Such isolates are probable contaminants. The specificity of the current definition of primary bloodstream infection could be increased by excluding enterococci mixed with skin organisms. Copyright © 2011 Association for Professionals in Infection Control and Epidemiology, Inc. Published by Mosby, Inc. All rights reserved.

  7. Predictive and prognostic factors in patients with blood-culture-positive community-acquired pneumococcal pneumonia.

    PubMed

    Amaro, Rosanel; Liapikou, Adamantia; Cilloniz, Catia; Gabarrus, Albert; Marco, Francesc; Sellares, Jacobo; Polverino, Eva; Garau, Javier; Ferrer, Miquel; Musher, Daniel M; Torres, Antoni

    2016-09-01

    In patients with pneumococcal community-acquired pneumonia (CAP), the risk factors for bacteraemia and its impact on outcomes are not fully elucidated. We aimed to compare characteristics of patients with blood-culture-positive versus blood-culture-negative pneumococcal CAP, and to characterise bacteraemic serotypes.We describe a prospective, observational study on nonimmunocompromised patients with pneumococcal CAP, from 1996 to 2013. We define severe pneumonia according to American Thoracic Society/Infectious Diseases Society of America guidelines.Of a total of 917 patients with pneumococcal CAP, 362 had blood-culture-positive pneumococcal pneumonia (BCPPP; 39%). High C-reactive protein (CRP) (≥20 mg·dL(-1)) (odds ratio (OR) 2.36, 95% CI 1.45-3.85), pleural effusion (OR 2.03, 95% CI 1.13-3.65) and multilobar involvement (OR 1.69, 95% CI 1.02-2.79) were independently associated with bacteraemic CAP, while nursing home resident (OR 0.12, 95% CI 0.01-1.00) was found as a protective factor. Despite the clinical differences, BCPPP showed similar outcomes to blood-culture-negative pneumococcal pneumonia (BCNPP). 14% of the serotypes (period 2006-2013) causing bacteraemia are included in pneumococcal conjugate vaccine PVC7, 74% in pneumococcal conjugate vaccine PVC13 and 83% in pneumococcal polysaccharide vaccine PPSV23.Pleural effusion, a high level of CRP and multilobar involvement predicted an increased risk of BCPPP. Although BCPPP patients were more severely ill at admission, mortality was not significantly greater than in BCNPP patients. Copyright ©ERS 2016.

  8. Human whole-blood culture system for ex vivo characterization of designer-cell function.

    PubMed

    Schukur, Lina; Geering, Barbara; Fussenegger, Martin

    2016-03-01

    Encapsulated designer cells implanted into mice are currently used to validate the efficacy of therapeutic gene networks for the diagnosis and treatment of various human diseases in preclinical research. Because many human conditions cannot be adequately replicated by animal models, complementary and alternative procedures to test future treatment strategies are required. Here we describe a novel approach utilizing an ex vivo human whole-blood culture system to validate synthetic biology-inspired designer cell-based treatment strategies. The viability and functionality of transgenic mammalian designer cells co-cultured with primary human immune cells were characterized. We demonstrated that transgenic mammalian designer cells required adequate insulation from the human blood microenvironment to maintain viability and functionality. The biomaterial alginate-(poly-l-lysine)-alginate used to encapsulate the transgenic designer cells did neither affect the viability of primary granulocytes and lymphocytes nor the functionality of lymphocytes. Additionally, alginate-encapsulated transgenic designer cells remained responsive to the release of the pro-inflammatory cytokine tumor necrosis factor (TNF) from the whole-blood culture upon exposure to bacterial lipopolysaccharide (LPS). TNF diffused into the alginate capsules, bound to the specific TNF receptors on the transgenic designer cells' surface and triggered the expression of the reporter gene SEAP (human placental secreted alkaline phosphatase) that was rewired to the TNF-specific signaling cascade. Human whole-blood culture systems can therefore be considered as valuable complementary assays to animal models for the validation of synthetic circuits in genetically modified mammalian cells and may speed up preclinical research in a world of personalized medicine.

  9. Detection of the Dimorphic Phases of Mucor circinelloides in Blood Cultures from an Immunosuppressed Female

    PubMed Central

    Arroyo, Miguel A.; Schmitt, Bryan H.; Davis, Thomas E.

    2016-01-01

    Mucormycosis fungemia is rarely documented since blood cultures are nearly always negative. We describe a case of Mucor circinelloides fungemia in a patient with a history of a sinus infection, sarcoidosis, and IgG deficiency. The identity of the isolate was supported by its microscopic morphology and its ability to convert into yeast forms under anaerobic conditions. The early detection, initiation of liposomal amphotericin B treatment, and reversal of underlying predisposing risk factors resulted in a good outcome. PMID:27777804

  10. Lack of requirement for blind subcultures of BACTEC blood culture media

    SciTech Connect

    McLaughlin, J.C.; Evers, J.L.; Officer, J.L.

    1981-11-01

    To determine the need for blind subculturing of BACTEC (Johnston Laboratories, Cockeysville, Md.) blood culture media, we compared results of radiometric readings, visual inspection, and blind subculturing for nearly 7,500 blood specimens. Visual inspection and radiometric testing were performed on day 1 through 7, and blind subcultures were made on day 3. In the first phase of the study, 402 of 3,896 aerobic bottles were positive by radiometric testing (growth index, greater than 25), visual inspection, or subculturing. Only six bottles were radiometrically negative but subculture positive on day 3. The second phase of the study was designed to determine if aerobic bottles eventually became radiometrically positive in those cases in which they were radiometrically negative but subculture positive on day 3. Two bottles were subculture positive but never gave a growth index of greater than or equal to 25 by day 7. One yielded Staphylococcus epidermidis, and one yielded viridans, Streptococcus sp. A total of 35 anaerobic organisms were isolated from 3,896 blood specimens. All of these anaerobes were detected by both radiometric testing and subculturing. We examined a total of 14,972 blood culture bottles. Twenty-nine bottles considered negative by visual inspection or radiometric readings were found to be positive by subculturing. Fifteen of these were shown, by chart review, to contain contaminants. Organisms in the other negative bottles would not have gone undetected because companion bottles from the same patients were radiometrically or visually positive. We concluded that it is necessary to perform blind subcultures of BACTEC 7B and 8B blood culture bottles.

  11. Sample taking during orthopedic surgery: sensitivity and specificity using the BACTEC blood culture system.

    PubMed

    Podleska, L E; Lendemans, S; Schmid, E; Hussmann, B; Nast-Kolb, D; Taeger, G

    2012-02-01

    The use of blood culture systems for sterile body fluids other than blood has proven to be superior to routine culture methods. This study was conducted in order to assess the performance of the BACTEC blood culture system compared to swab/tissue sample collection for the detection of infection from intraoperative samples taken during surgical procedures. Sensitivity was determined by taking samples (BACTEC and swab/tissue samples) from patients with clinically evident infection (Infection group). Specificity was tested by taking the same sample sets from patients who had aseptic operations with no history of infection (Control group). The sensitivity was found to be much higher for the BACTEC group (50 isolates from 56 samples, sensitivity: 89%) compared to the swab/tissue samples (29 isolates out of 56 samples, sensitivity: 52%). The specificity was lower in the BACTEC group (32 isolates out of 44 samples, specificity: 27%) compared to the swab/tissue samples (1 isolate out of 44 samples, specificity: 98%). We conclude that BACTEC is useful for intraoperative sample collection in cases of low-grade infection. However, it is less specific and there is always the possibility for contamination. Therefore, it is advisable to use this technique in combination with regular tissue samples.

  12. [Disseminated mycobacterial infections in patients with HIV/AIDS. Evaluation of blood cultures].

    PubMed

    Coitinho, C; Brandes, E; Pardiñas, M; Rivas, C

    2005-01-01

    One thousand-forty blood cultures corresponding to 451 Uruguayan patients with AIDS and clinic diagnosis of disseminated mycobacterial infection were evaluated between 1999 and 2003. Samples were processed in the National Reference Center for Mycobacteria (Montevideo, Uruguay), using the automated blood culture system for mycobacteria MB-BacT (BioMérieux). Forty-five positive samples were detected (4.3%) corresponding to 26 patients with AIDS (average 2.3 samples per patient). In 10/26 patients M. avium complex (MAC) was identified and in 13/26 the isolated germ was M. tuberculosis. The average time of incubation was of 12.4 days (range 6-19 days) for MAC and of 22.6 days (range 7-35 days) for M. tuberculosis. Blood culture has demonstrated to be the best sample for the bacteriological confirmation of the disseminated mycobacterial infections when at least 2 samples by patient are studied. The frequency of isolates of M. tuberculosis and MAC in AIDS patients is according with a moderate prevalence of tuberculosis in Uruguay.

  13. Acoustic trapping for bacteria identification in positive blood cultures with MALDI-TOF MS.

    PubMed

    Hammarström, Björn; Nilson, Bo; Laurell, Thomas; Nilsson, Johan; Ekström, Simon

    2014-11-04

    Matrix assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) is currently changing the clinical routine for identification of microbial pathogens. One important application is the rapid identification of bacteria for the diagnosis of bloodstream infections (BSI). A novel approach based on acoustic trapping and an integrated selective enrichment target (ISET) microchip that improves the sample preparation step for this type of analysis is presented. The method is evaluated on clinically relevant samples in the form of Escherichia coli infected blood cultures. It is shown that noncontact acoustic trapping enables capture, enrichment, and washing of bacteria directly from the complex background of crude blood cultures. The technology replaces centrifugation-based separation with a faster and highly automated sample preparation method that minimizes manual handling of hazardous pathogens. The presented method includes a solid phase extraction step that was optimized for enrichment of the bacterial proteins and peptides that are used for bacterial identification. The acoustic trapping-based assay provided correct identification in 12 out 12 cases of E. coli positive blood cultures with an average score of 2.19 ± 0.09 compared to 1.98 ± 0.08 when using the standard assay. This new technology opens up the possibility to automate and speed up an important and widely used diagnostic assay for bloodstream infections.

  14. Coagulase-negative staphylococci strains resistant to oxacillin isolated from neonatal blood cultures.

    PubMed

    Pereira, Valéria Cataneli; Cunha, Maria de Lourdes Ribeiro de Souza da

    2013-11-01

    Coagulase-negative staphylococci (CoNS) are the microorganisms most frequently isolated from clinical samples and are commonly found in neonatal blood cultures. Oxacillin is an alternative treatment of choice for CoNS infections; however, resistance to oxacillin can have a substantial impact on healthcare by adversely affecting morbidity and mortality. The objective of this study was to detect and characterise oxacillin-resistant CoNS strains in blood cultures of newborns hospitalised at the neonatal ward of the University Hospital of the Faculty of Medicine of Botucatu. One hundred CoNS strains were isolated and the mecA gene was detected in 69 of the CoNS strains, including 73.2% of Staphylococcus epidermidis strains, 85.7% of Staphylococcus haemolyticus strains, 28.6% of Staphylococcus hominis strains and 50% of Staphylococcus lugdunensis strains. Among these oxacillin-resistant CoNS strains, staphylococcal cassette chromosome mec (SCCmec) type I was identified in 24.6%, type II in 4.3%, type III in 56.5% and type IV in 14.5% of the strains. The data revealed an increase in the percentage of CoNS strains isolated from blood cultures from 1991-2009. Furthermore, a predominant SCCmec profile of the oxacillin-resistant CoNS strains isolated from neonatal intensive care units was identified with a prevalence of SCCmec types found in hospital-acquired strains.

  15. [Effectiveness of intervention by the infection control team for cancer patients with a positive blood culture].

    PubMed

    Yamada, Tomoyuki; Suzuki, Kaoru; Ohi, Yukimasa; Kawanishi, Fumiko; Shibata, Yuriko; Hosomi, Makoto; Goto, Emi; Nishihara, Masami; Katsumata, Takahiro; Ukimura, Akira

    2013-11-01

    Cancer patients at a high risk of acquiring infectious diseases should be maintained in a facility where good infection control practices are followed. At our hospital, the infection control team(ICT)provides expertise, education, and support to the staff, helping them maintain proper standards, thereby minimizing the risks of infection. The ICT(established in 2004)has implemented infection control programs by employing an appropriate number of staff members after the revision of medical treatment fees in 2011. Our intervention program includes 2 general policies, namely, ordering and collection of blood cultures and intervention for the medical care of patients with positive blood cultures. In this study, we evaluated the effectiveness of our intervention for cancer patients with a positive blood culture. During the surveillance period(April 2011 to July 2012), 42 positive cases were determined to be infectious. ICT intervention was required in 37 cases. Our suggestions were accepted in 92%(34/37)of the cases, and improved outcome was estimated in 65%(22/34)of the cases. The results of our study contribute to the scientific bases on which routine clinical practices could be promoted in the future.

  16. Multi-Probe Real-Time PCR Identification of Common Mycobacterium Species in Blood Culture Broth

    PubMed Central

    Foongladda, Suporn; Pholwat, Suporn; Eampokalap, Boonchuay; Kiratisin, Pattarachai; Sutthent, Ruengpung

    2009-01-01

    Mycobacterium tuberculosis complex, M. avium, and M. intracellulare are the most common causes of systemic bacterial infection in AIDS patients. To identify these mycobacterial isolates in primary blood culture broths, we developed a multiple hybridization probe-based real-time PCR assay using the LightCycler system. The primers were designed to amplify a 320-bp fragment of Mycobacterium 16S rRNA genes. Reaction specificity was evaluated using PCR amplification curves along with specific melting temperatures of probes on DNA extracted from 13 Mycobacterium species. In this study, results showed 100% accuracy for the selected bacterial panel. Detection limits were 350, 600, and 650 colony-forming unit (CFU)/ml blood culture broths for M. tuberculosis complex, M. avium, and M. intracellulare, respectively (1 to 2 CFU/reaction). To evaluate clinical applicability, 341 acid-fast bacilli in blood culture broths were analyzed. In total, 327 (96%) were positively identified: 54.5% M. tuberculosis complex, 37.5% M. avium, and 3.8% M. intracellulare. Results can be available within 3 hours of receiving a broth sample, which makes this rapid and simple assay an attractive diagnostic tool for clinical use. PMID:19095775

  17. Sterility testing of a total nutrient admixture with a biphasic blood-culture system.

    PubMed

    Murray, P R; Sandrock, M J

    1991-11-01

    The ability of a biphasic blood-culture system to detect microbial contamination of a total nutrient admixture (TNA) was studied. A TNA consisting of amino acids, dextrose, lipid emulsion, electrolytes, and trace elements was prepared under aseptic conditions. Septi-Chek blood-culture bottles were then injected with a TNA sample and with one of various dilutions of five bacterial or fungal suspensions, and an agar slide unit was attached to each bottle. One bottle was injected only with a TNA sample. The bottles were incubated and examined once daily. Each bottle was inverted immediately after inoculation and at each observation time to allow the broth to wash over the agar slide unit. Because the lipid contained in the TNA caused the broth in all bottles to become turbid, it was impossible to determine whether microbial growth was present in the broth. Microbial growth, however, became evident in 24-72 hours on all slide units except the one attached to the bottle that had been injected only with TNA. The Septi-Chek biphasic blood-culture system appears to be useful in evaluating the sterility of TNAs.

  18. Rapid identification of bacteria, mecA and van genes from blood cultures.

    PubMed

    Prère, M-F; Baron, O; Fayet, O

    2007-11-01

    The Genotype technology, a quick molecular genetic assay based on DNA multiplex amplification with biotinylated primers followed by hybridization to membrane bound probes, complies with the requirements for a fast diagnosis of sepsis. We evaluated the new Genotype BC Gram-negative and Gram-positive test kits (Hain Life Science, Germany) which respectively allow for the identification of 15 species of Gram-negative (GN) rods, and the identification of 17 Gram-positive (GP) bacteria species together with the determination of methicillin and vancomycin resistance (mecA and van genes). The study was performed on 60 positive blood cultures from BacT/ALERT bottles (aerobic, anaerobic and pediatric bottles). First, a Gram stain was carried out to select between Genotype BC GP or GN test, then identification were performed by the Genotype BC tests and by biochemical conventional tests after subculture and phenotypic susceptibility determination. The operating procedure was very easy to carry out and required a small amount of starting material (5 to 10 microL of blood culture). The results were available within 4.5 hours. For all the blood cultures, the Genotype BC results correlated with the biochemical identification and phenotypic antibiotics susceptibility. According to our results, this DNA strip technology based assay can easily be incorporated into routine diagnosis.

  19. Two-year experience with aerobic culturing of apheresis and whole blood-derived platelets.

    PubMed

    Kleinman, Steven H; Kamel, Hany T; Harpool, Dennis R; Vanderpool, Sandra K; Custer, Brian; Wiltbank, Thomas B; Nguyen, Kim-Anh; Tomasulo, Peter A

    2006-10-01

    Throughout its system of regional centers, Blood Systems implemented culture based bacterial testing with a standardized protocol for both apheresis and whole blood-derived platelets (PLTs). After a 24-hour hold, 4 mL of PLT product was inoculated into an aerobic bottle (BacT/ALERT, bioMérieux). Cultures were incubated for 24 hours before routine product release to prevent distribution of infected products while minimizing consignee notification, product retrievals, and hospital PLT inventory problems. Initial-positives were further tested (and bacteria identified) by performing cultures from the original component and subcultures from the BacT/ALERT bottle. Results were categorized according to AABB recommended definitions with minor modifications. The rate of true-positive detections from culturing 122,971 apheresis PLTs was 0.017 percent (95% confidence interval [CI], 0.011%-0.026%). All true-positive microorganisms were Gram-positive with a predominance of coagulase-negative Staphylococcus and Bacillus species. Twenty of the 21 true-positive samples (95%) were detected by 24 hours but only 14 (68%) were detected by 18 hours. The false-positive rate due to contamination was 0.1 percent with the majority of isolates being skin or environmental organisms. Results did not differ significantly for whole blood-derived versus apheresis PLTs. These data corroborate the fact that the rate of detection of truly contaminated PLT apheresis products in the United States is approximately 1 in 5000 (0.02%); this is lower than the 0.03 to 0.05 percent rates that were generally quoted in the literature before the implementation of prospective bacterial culturing programs.

  20. [A case with Abiotrophia defectiva isolated from postoperative blood culture of a patient with diffuse peritonitis].

    PubMed

    Tanamachi, Chiyoko; Hashimoto, Kouji; Oyama, Nana; Horita, Rie; Yano, Tomomi; Tashiro, Naotaka; Fuyuta, Syuhei; Sakamoto, Teruo; Nakashima, Osamu

    2012-08-01

    We have recently detected Abiotrophia defectiva (A. defectiva) from postoperative blood culture of a patient with diffuse peritonitis. Pleomorphic Gram positive rod appearance and no growth on a standard sheep blood agar led us to suspect aerotolerant anaerobe. From the test results of simplified identification kit for anaerobes, we tentatively reported Clostridium tertium (C. tertium) to the clinician. However, the analysis of 16SrRNA sequence proved it to be A. defectiva. When Gram positive coccus or pleomorphic Gram positive rod are observed on blood culture, it is recommended to incubate Brucella HK (RS) blood agar plates for 24 hours at 35 degrees C both aerobically and anaerobically. The growth both aerobically and anaerobically suggests A. defectiva and Satellitism test and Rapid ID 32 STREP(SYSMEX) testing should be performed to identify the responsible bacteria. A. defectiva is Gram positive streptococcus and requires pyridoxal hydrochloride or L-cysteine to grow. It could be unidentified or misidentified and its particularity often leads incomplete report to the clinician. Our study shows that more careful examination will increase the detection of A. defectiva.

  1. Reducing time to identification of aerobic bacteria and fastidious micro-organisms in positive blood cultures.

    PubMed

    Intra, J; Sala, M R; Falbo, R; Cappellini, F; Brambilla, P

    2016-12-01

    Rapid and early identification of micro-organisms in blood has a key role in the diagnosis of a febrile patient, in particular, in guiding the clinician to define the correct antibiotic therapy. This study presents a simple and very fast method with high performances for identifying bacteria by matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) after only 4 h of incubation. We used early bacterial growth on PolyViteX chocolate agar plates inoculated with five drops of blood-broth medium deposited in the same point and spread with a sterile loop, followed by a direct transfer procedure on MALDI-TOF MS target slides without additional modification. Ninety-nine percentage of aerobic bacteria were correctly identified from 600 monomicrobial-positive blood cultures. This procedure allowed obtaining the correct identification of fastidious pathogens, such as Streptococcus pneumoniae, Neisseria meningitidis and Haemophilus influenzae that need complex nutritional and environmental requirements in order to grow. Compared to the traditional pathogen identification from blood cultures that takes over 24 h, the reliability of results, rapid performance and suitability of this protocol allowed a more rapid administration of optimal antimicrobial treatment in the patients.

  2. Clinical laboratory comparison of lysis-centrifugation and BACTEC radiometric blood culture techniques.

    PubMed

    McLaughlin, J C; Hamilton, P; Scholes, J V; Bartlett, R C

    1983-11-01

    The lysis-centrifugation technique (ISOLATOR; E. I. du Pont de Nemours & Co., Wilmington, Del.) and the radiometric blood culture technique (BACTEC; Johnston Laboratories, Inc., Cockeysville, Md.) were compared on 1,000 blood cultures. A total of 16 ml of blood was distributed: 8 ml into an ISOLATOR 7.5 microbial tube and 4 ml each into BACTEC 7C and 8B bottles. The concentrate from the ISOLATOR tubes was inoculated under a laminar-flow hood onto two sheep blood agar plates (one incubated in CO2 and one incubated anaerobically), one chocolate agar plate, and one brain heart infusion agar plate. Of 91 blood specimens obtained that yielded clinically significant organisms, 52 were positive by both systems, 27 were positive by the ISOLATOR system only, and 12 were positive by the BACTEC system only. From the positive blood specimens, 97 clinically significant organisms were isolated: 57 by both systems, 27 by the ISOLATOR system only, and 13 by the BACTEC system only. Of the 57 organisms detected by both systems, 28 were detected simultaneously, 13 were detected earlier by the ISOLATOR system, and 16 were detected earlier by the BACTEC system. Isolated colonies were obtained earlier by the ISOLATOR system in 40 cases and by the BACTEC system in 5 cases. Organisms determined to be contaminants by thorough chart review were isolated from 138 ISOLATOR tubes. In 98 instances, these were represented by one colony of Staphylococcus epidermidis, alpha-hemolytic streptococci, or diphtheroids. The ability to determine CFU per milliliter with the ISOLATOR system did not help differentiate clinically significant organisms from contaminants.

  3. Highly Sensitive Bacteriophage-Based Detection of Brucella abortus in Mixed Culture and Spiked Blood

    PubMed Central

    Sergueev, Kirill V.; Filippov, Andrey A.; Nikolich, Mikeljon P.

    2017-01-01

    For decades, bacteriophages (phages) have been used for Brucella species identification in the diagnosis and epidemiology of brucellosis. Traditional Brucella phage typing is a multi-day procedure including the isolation of a pure culture, a step that can take up to three weeks. In this study, we focused on the use of brucellaphages for sensitive detection of the pathogen in clinical and other complex samples, and developed an indirect method of Brucella detection using real-time quantitative PCR monitoring of brucellaphage DNA amplification via replication on live Brucella cells. This assay allowed the detection of single bacteria (down to 1 colony-forming unit per milliliter) within 72 h without DNA extraction and purification steps. The technique was equally efficient with Brucella abortus pure culture and with mixed cultures of B. abortus and α-proteobacterial near neighbors that can be misidentified as Brucella spp., Ochrobactrum anthropi and Afipia felis. The addition of a simple short sample preparation step enabled the indirect phage-based detection of B. abortus in spiked blood, with the same high sensitivity. This indirect phage-based detection assay enables the rapid and sensitive detection of live B. abortus in mixed cultures and in blood samples, and can potentially be applied for detection in other clinical samples and other complex sample types. PMID:28604602

  4. Highly Sensitive Bacteriophage-Based Detection of Brucella abortus in Mixed Culture and Spiked Blood.

    PubMed

    Sergueev, Kirill V; Filippov, Andrey A; Nikolich, Mikeljon P

    2017-06-10

    For decades, bacteriophages (phages) have been used for Brucella species identification in the diagnosis and epidemiology of brucellosis. Traditional Brucella phage typing is a multi-day procedure including the isolation of a pure culture, a step that can take up to three weeks. In this study, we focused on the use of brucellaphages for sensitive detection of the pathogen in clinical and other complex samples, and developed an indirect method of Brucella detection using real-time quantitative PCR monitoring of brucellaphage DNA amplification via replication on live Brucella cells. This assay allowed the detection of single bacteria (down to 1 colony-forming unit per milliliter) within 72 h without DNA extraction and purification steps. The technique was equally efficient with Brucella abortus pure culture and with mixed cultures of B. abortus and α-proteobacterial near neighbors that can be misidentified as Brucella spp., Ochrobactrum anthropi and Afipia felis. The addition of a simple short sample preparation step enabled the indirect phage-based detection of B. abortus in spiked blood, with the same high sensitivity. This indirect phage-based detection assay enables the rapid and sensitive detection of live B. abortus in mixed cultures and in blood samples, and can potentially be applied for detection in other clinical samples and other complex sample types.

  5. Isolation of Corynebacterium tuscaniae sp. nov. from Blood Cultures of a Patient with Endocarditis

    PubMed Central

    Riegel, Philippe; Creti, Roberta; Mattei, Romano; Nieri, Alfredo; von Hunolstein, Christina

    2006-01-01

    A strain of an unknown coryneform bacterium was repeatedly isolated in pure culture from the blood of a patient affected by endocarditis. Comparative 16S rRNA gene sequence analysis revealed that this isolate represented a new subline within the genus Corynebacterium. This new taxon can be identified by the presence of corynomycolic acids and its enzymatic activities and fermentation of sugars. Acid production from glucose and maltose, pyrazinamidase and alkaline phoshatase activities, and hippurate hydrolysis were the most characteristic phenotypic features of the bacterium. On the basis of both phenotypic and phylogenetic evidence, it is proposed that this isolate be classified as a novel species, Corynebacterium tuscaniae sp. nov. The type strain, ISS-5309, has been deposited in the American Type Culture Collection (ATCC BAA-1141) and in the Culture Collection of the University of Göteborg (CCUG 51321). PMID:16455875

  6. Antibiotic susceptibility testing of grown blood cultures by combining culture and real-time polymerase chain reaction is rapid and effective.

    PubMed

    Beuving, Judith; Verbon, Annelies; Gronthoud, Firza A; Stobberingh, Ellen E; Wolffs, Petra F G

    2011-01-01

    Early administration of appropriate antibiotic therapy in bacteraemia patients dramatically reduces mortality. A new method for RApid Molecular Antibiotic Susceptibility Testing (RAMAST) that can be applied directly to positive blood cultures was developed and evaluated. Growth curves and antibiotic susceptibility of blood culture isolates (Staphylococcus aureus, enterococci and (facultative) aerobic gram-negative rods) were determined by incubating diluted blood cultures with and without antibiotics, followed by a quantitative universal 16S PCR to detect the presence or absence of growth. Testing 114 positive blood cultures, RAMAST showed an agreement with microbroth dilution of 96.7% for gram-negative rods, with a minor error (false-susceptibility with a intermediate resistant strain) rate of 1.9%, a major error (false resistance) rate of 0.8% and a very major error (false susceptibility) rate of 0.6%. Agreement for S. aureus was 97.9%, with a very major error rate of 2.1%. Enterococcus species showed 95.0% agreement, with a major error rate of 5.0%. These agreements are comparable with those of the Phoenix system. Starting from a positive blood culture, the test was completed within 9 hours. This new rapid method for antibiotic susceptibility testing can potentially provide accurate results for most relevant bacteria commonly isolated from positive blood cultures in less time than routine methods.

  7. Blood

    MedlinePlus

    ... The liquid part, called plasma, is made of water, salts, and protein. Over half of your blood is plasma. The solid part of your blood contains red blood cells, white blood cells, and platelets. Red ...

  8. Rapid detection of positive blood cultures with the BACTEC NR-660 does not require first-day subculturing.

    PubMed Central

    Levi, M H; Gialanella, P; Motyl, M R; McKitrick, J C

    1988-01-01

    An analysis of blood culture data was performed to determine whether subculturing within the first 24 h of incubation decreased the time to detection of positive blood cultures when compared with the routine use of the BACTEC NR-660 system (Johnston Laboratories, Inc., Towson, Md.). During a 9-month period (June 1985 to February 1986), 17,913 blood cultures were received in our laboratory, of which 1,463 (8.2%) became positive. Of the positive cultures, 97% were detected with equal or greater rapidity by the NR-660 system than by visual inspection and first-day blind subculturing. There were 37 delayed positive cultures from which only one isolate (0.07%) was not eventually detected by the NR-660 system. Coagulase-negative staphylococcus was the most frequent isolate among the delayed positive cultures, but only 3 of 15 isolates were known to be clinically significant isolates. The longest delay in detection by the NR-660 system was 6 days for one isolate of Cryptococcus neoformans and one isolate of Klebsiella pneumoniae. Although subculturing may decrease the time to detection of a few cultures, the majority of positive blood cultures were detected faster or with equal speed by the NR-660 system. When the data were evaluated, routine use of the NR-660 system was sufficient for the detection of positive blood cultures and was cost-effective. PMID:3069859

  9. Performance of the Verigene Gram-negative blood culture assay for rapid detection of bacteria and resistance determinants.

    PubMed

    Dodémont, Magali; De Mendonça, Ricardo; Nonhoff, Claire; Roisin, Sandrine; Denis, Olivier

    2014-08-01

    Nonduplicate blood cultures that were positive for Gram-negative bacilli (n = 125) were tested by the Verigene Gram-negative blood culture (BC-GN) assay; 117 (90.7%) isolates were members of the panel. For identification and resistance markers, the agreements with routine methods were 97.4% (114/117) and 92.3% (12/13). The BC-GN assay is a rapid and accurate tool for the detection of pathogens from blood cultures and could be integrated alongside conventional systems to enable faster patient management, but the clinical benefits should be further evaluated.

  10. Effects of Culture Conditions and Tuberculin Source on Interferon-gamma production in Whole Blood Cultures from Mycobacterium bovis Infected Cattle

    USDA-ARS?s Scientific Manuscript database

    The BOVIGAM® interferon (IFN) - gamma assay constitutes an ante-mortem, in vitro laboratory-based tuberculosis test and is used complementary to the tuberculin skin test. The assay is performed in two stages: firstly, whole blood is cultured with antigens stimulating blood leucocytes to produce IFN...

  11. Effects of Culture Conditions and Tuberculin Source on Interferon-gamma Production in Whole Blood Cultures from Mycobacterium bovis Infected Cattle

    USDA-ARS?s Scientific Manuscript database

    The BOVIGAM® interferon (IFN) - gamma assay constitutes an ante-mortem, in vitro laboratory-based tuberculosis test and is used complementary to the tuberculin skin test. The assay is performed in two stages: firstly, whole blood is cultured with antigens stimulating blood leucocytes to produce IFN-...

  12. Acceleration of Antimicrobial Susceptibility Testing of Positive Blood Cultures by Inoculation of Vitek 2 Cards with Briefly Incubated Solid Medium Cultures

    PubMed Central

    Idelevich, Evgeny A.; Schüle, Isabel; Grünastel, Barbara; Wüllenweber, Jörg; Peters, Georg

    2014-01-01

    Briefly incubated agar cultures from positive blood cultures were used for antimicrobial susceptibility testing (AST) by Vitek 2. The cultivation time until inoculation was 3.8 h for Gram-positive cocci and 2.4 h for Gram-negative rods. The error rates were low, providing early and reliable AST without additional time or cost expenditure. PMID:25165084

  13. Acceleration of antimicrobial susceptibility testing of positive blood cultures by inoculation of Vitek 2 cards with briefly incubated solid medium cultures.

    PubMed

    Idelevich, Evgeny A; Schüle, Isabel; Grünastel, Barbara; Wüllenweber, Jörg; Peters, Georg; Becker, Karsten

    2014-11-01

    Briefly incubated agar cultures from positive blood cultures were used for antimicrobial susceptibility testing (AST) by Vitek 2. The cultivation time until inoculation was 3.8 h for Gram-positive cocci and 2.4 h for Gram-negative rods. The error rates were low, providing early and reliable AST without additional time or cost expenditure.

  14. Using qualitative research methods in biomedical innovation: the case of cultured red blood cells for transfusion.

    PubMed

    Lyall, Catherine; King, Emma

    2016-05-11

    Qualitative research has a key role to play in biomedical innovation projects. This article focuses on the appropriate use of robust social science methodologies (primarily focus group studies) for identifying the public's willingness and preference for emerging medical technologies. Our study was part of the BloodPharma project (now known as the Novosang project) to deliver industrially generated red blood cells for transfusion. Previous work on blood substitutes shows that the public prefers donated human blood. However, no research has been conducted concerning attitudes to stem cell derived red blood cells. Qualitative research methods including interviews and focus groups provide the methodological context for this paper. Focus groups were used to elicit views from sub-sections of the UK population about the potential use of such cultured red blood cells. We reflect on the appropriateness of that methodology in the context of the BloodPharma project. Findings are in the form of lessons transferable to other interdisciplinary, science-led teams about what a social science dimension can bring; why qualitative research should be included; and how it can be used effectively. Qualitative data collection offers the strength of exploring ambivalence and investigating the reasons for views, but not necessarily their prevalence in wider society. The inherent value of a qualitative method, such as focus groups, therefore lies in its ability to uncover new information. This contrasts with a quantitative approach to simply 'measuring' public opinion on a topic about which participants may have little prior knowledge. We discuss a number of challenges including: appropriate roles for embedded social scientists and the intricacies of doing upstream engagement as well as some of the design issues and limitations associated with the focus group method.

  15. [Stimulation of cell cultures recovery after cryopreservation by the cattle cord blood FRACTION (below 5 kDa) or Actovegin].

    PubMed

    Gulevskiĭ, A K; Trifonova, A V; Lavrik, A A

    2013-01-01

    The capacities of the cattle cord blood low-molecular fraction (below 5 kDa) and Actovegin (the vealer blood fraction (below 5 kDa)) for recovering functions of cell cultures after cryopreservation compared. Their influence proliferation of the flozen-thawed cell cultures, certain stages of their growth, cell attachment, rate of cell spreading, and mitotic regiment has been studied. Both the cord blood low-molecular fraction and Actovegin were shown to stimulate growth of the cell cultures after cryopreservation more efficiently at the concentration of 224 μg/ml. However, despite the stimulating effect discovered, their application did not bring proliferative indices on the 1st passage after cryopreservation to the values of the native culture. The effects of the cord blood low-molecular fraction and Actovegin on the human fibroblast culture were identical by the following parameters: cell attachment, rates of cell spreading and proliferation. In culture BHK-21 clone 13/04 the efficiency of Actovegin was low, while the cord blood low-molecular fraction has a conspicuous stimulating effect on its adhesion and proliferation. The investigations carried out can serve as a basis for the development of regenerative media containing the cattle cord blood low-molecular fraction (below 5 kDa) or Actovegin as active components at the concentration of 224 μg/ml with the purpose of fast recovery of culture prolifetative properties after cryopreservation.

  16. Blood culture and polymerase chain reaction for the diagnosis of the chronic phase of human infection with Trypanosoma cruzi.

    PubMed

    Castro, A M; Luquetti, A O; Rassi, A; Rassi, G G; Chiari, E; Galvão, L M C

    2002-10-01

    In the present study, we evaluated for the first time the profile of blood parasitism in untreated, chronic Chagas' disease. The study was conducted on 60 patients and a control group of nine serologically negative individuals. Analysis of three blood samples showed 70% cumulative positivity for blood culture and 86.7% positivity for PCR. The comparison of the two tests revealed that 41.1% (74/180) of the samples presented positive results for both PCR and blood culture, 22.2% (40/180) were positive for PCR alone, and 4.4% (8/180) were positive for blood culture and negative for PCR. The addition of the second sample raised positivity significantly for both blood culture ( P=0.0000) and PCR ( P=0.0369). Addition of the third sample was also statistically significant for blood culture ( P=0.0001) but not for PCR ( P=0.1186). These data point to the importance of studying the parasitemia of Trypanosoma cruzi-infected individuals before specific treatment. They also suggest that at least two blood samples should be collected and that two tests should be used, if possible--a procedure that considerably improves the parasitologic diagnosis of Chagas' disease and the evaluation of therapeutic efficacy.

  17. Performance of the FilmArray® blood culture identification panel utilized by non-expert staff compared with conventional microbial identification and antimicrobial resistance gene detection from positive blood cultures.

    PubMed

    McCoy, Morgan H; Relich, Ryan F; Davis, Thomas E; Schmitt, Bryan H

    2016-07-01

    Utilization of commercially available rapid platforms for microbial identification from positive blood cultures is useful during periods of, or in laboratories with, limited expert staffing. We compared the results of the FilmArray® BCID Panel performed by non-expert technologists to those of conventional methods for organism identification performed by skilled microbiologists. Within 8 h of signalling positive by a continuous monitoring blood culture system, positive bottles were analysed by the FilmArray BCID Panel. Data from these analyses were compared to standard-of-care testing, which included conventional and automated methods. To gauge the ease of use of the BCID Panel by non-expert staff, technologists unfamiliar with diagnostic bacteriology performed the testing without prior knowledge of the Gram stain results, or even whether organisms were detected. Identifications of 172/200 (86 %) positive blood cultures using the BCID Panel were consistent with identifications provided by standard-of-care methods. Standard-of-care testing identified organisms in 20 positive blood cultures, which were not represented on the BCID Panel. Seven (3.5 %) blood cultures demonstrated a discrepancy between the methods, which could not be attributed to either a lack of representation on the panel or unclear separate detection of organisms in a mixed blood culture of a shared genus or grouping of organisms, e.g. Staphylococcus or Enterobacteriaceae . One (0.5 %) blood culture yielded invalid results on two separate panels, so it was eliminated from the study. The easy-to-use FilmArray® technology shows good correlation with blood culture identification and antibiotic resistance detection performed by conventional methods. This technology may be particularly useful in laboratories with limited staffing or limited technical expertise.

  18. Purification and microRNA profiling of exosomes derived from blood and culture media.

    PubMed

    McDonald, Marguerite K; Capasso, Kathryn E; Ajit, Seena K

    2013-06-14

    Stable miRNAs are present in all body fluids and some circulating miRNAs are protected from degradation by sequestration in small vesicles called exosomes. Exosomes can fuse with the plasma membrane resulting in the transfer of RNA and proteins to the target cell. Their biological functions include immune response, antigen presentation, and intracellular communication. Delivery of miRNAs that can regulate gene expression in the recipient cells via blood has opened novel avenues for target intervention. In addition to offering a strategy for delivery of drugs or RNA therapeutic agents, exosomal contents can serve as biomarkers that can aid in diagnosis, determining treatment options and prognosis. Here we will describe the procedure for quantitatively analyzing miRNAs and messenger RNAs (mRNA) from exosomes secreted in blood and cell culture media. Purified exosomes will be characterized using western blot analysis for exosomal markers and PCR for mRNAs of interest. Transmission electron microscopy (TEM) and immunogold labeling will be used to validate exosomal morphology and integrity. Total RNA will be purified from these exosomes to ensure that we can study both mRNA and miRNA from the same sample. After validating RNA integrity by Bioanalyzer, we will perform a medium throughput quantitative real time PCR (qPCR) to identify the exosomal miRNA using Taqman Low Density Array (TLDA) cards and gene expression studies for transcripts of interest. These protocols can be used to quantify changes in exosomal miRNAs in patients, rodent models and cell culture media before and after pharmacological intervention. Exosomal contents vary due to the source of origin and the physiological conditions of cells that secrete exosomes. These variations can provide insight on how cells and systems cope with stress or physiological perturbations. Our representative data show variations in miRNAs present in exosomes purified from mouse blood, human blood and human cell culture media

  19. Early detection and preliminary susceptibility testing of positive pediatric blood cultures with the Steers replicator.

    PubMed Central

    Paisley, J W; Todd, J K; Roe, M H

    1977-01-01

    Early replicator subculturing of blood specimens after 4 to 16 h of incubation detected 135 of 217 (59%) of all positive cultures, including 56 of 64 (87%) Haemophilus influenzae type b, 7 of 22 (32%) Staphylococcus aureus, 19 of 20 (95%) pathogenic streptococci, and 20 of 24 (83%) Enterobacteriaceae. The mean time to detection of the common pediatric pathogens (36 h) was significantly less than that of contaminants (85 h) (P less than 0.001). Blind subculturing to differential media aided in the rapid identification of isolates and the detection of mixed cultures. In addition, a method of obtaining rapid susceptibilities of blood and body fluid isolates to selected antibiotics by blind subculturing to antibiotic-containing media was evaluated. Immediate susceptibility information was obtained for 214 of the 245 (87.3%) isolate-antibiotic combinations. There was complete correlation with a standard Kirby-Bauer reading for 94.9% of these observations. Replicator blood subculturing before 24 h of incubation results in early detection of the majority of pediatric pathogens. The inoculation of additional differential and antibiotic-containing media with each blind subculture aids in rapid identification of isolates and may give limited, but clinically important, immediate susceptibility information. Images PMID:914993

  20. Early detection and preliminary susceptibility testing of positive pediatric blood cultures with the Steers replicator.

    PubMed

    Paisley, J W; Todd, J K; Roe, M H

    1977-10-01

    Early replicator subculturing of blood specimens after 4 to 16 h of incubation detected 135 of 217 (59%) of all positive cultures, including 56 of 64 (87%) Haemophilus influenzae type b, 7 of 22 (32%) Staphylococcus aureus, 19 of 20 (95%) pathogenic streptococci, and 20 of 24 (83%) Enterobacteriaceae. The mean time to detection of the common pediatric pathogens (36 h) was significantly less than that of contaminants (85 h) (P less than 0.001). Blind subculturing to differential media aided in the rapid identification of isolates and the detection of mixed cultures. In addition, a method of obtaining rapid susceptibilities of blood and body fluid isolates to selected antibiotics by blind subculturing to antibiotic-containing media was evaluated. Immediate susceptibility information was obtained for 214 of the 245 (87.3%) isolate-antibiotic combinations. There was complete correlation with a standard Kirby-Bauer reading for 94.9% of these observations. Replicator blood subculturing before 24 h of incubation results in early detection of the majority of pediatric pathogens. The inoculation of additional differential and antibiotic-containing media with each blind subculture aids in rapid identification of isolates and may give limited, but clinically important, immediate susceptibility information.

  1. Ability of procalcitonin to diagnose bacterial infection and bacteria types compared with blood culture findings

    PubMed Central

    Watanabe, Yuji; Oikawa, Nozomi; Hariu, Maya; Fuke, Ryota; Seki, Masafumi

    2016-01-01

    Procalcitonin (PCT) and C-reactive protein serve as biomarkers of infection in patients with sepsis/bacteremia. The present study assessed the clinical characteristics of 280 patients with suspected sepsis who were admitted to Tohoku Medical and Pharmaceutical University Hospital between January 2012 and December 2013. Among the patients, 133 and 147 were positive and negative for PCT, respectively. Patients who were PCT positive were older and more frequently male, had reduced levels of platelets and albumin, and increased levels of aspartate aminotransferase, alanine aminotransferase, blood urea nitrogen, creatinine, and C-reactive protein. Patients who were PCT positive had significantly higher blood culture positivity compared with those who were PCT negative, and the sensitivity and specificity of PCT for detecting positive blood cultures were 74.5% and 59.1%, respectively. Escherichia coli was detected in PCT-positive patients, whereas Staphylococcus epidermidis and Staphylococcus lugdunensis were frequently detected in PCT-negative patients. Levels of PCT were higher in the patients infected with gram-negative rods than those with gram-positive cocci. Furthermore, extended-spectrum β-lactamase (ESBL)-producing bacteria cases showed higher levels of PCT than those of non-ESBL cases. These results suggest that PCT may be a useful biomarker of sepsis, and it might serve as a strong tool to detect patients with severe gram-negative rod bacteremia including ESBL-producing bacteria cases early due to its relative high sensitivity. PMID:27757046

  2. Ability of procalcitonin to diagnose bacterial infection and bacteria types compared with blood culture findings.

    PubMed

    Watanabe, Yuji; Oikawa, Nozomi; Hariu, Maya; Fuke, Ryota; Seki, Masafumi

    2016-01-01

    Procalcitonin (PCT) and C-reactive protein serve as biomarkers of infection in patients with sepsis/bacteremia. The present study assessed the clinical characteristics of 280 patients with suspected sepsis who were admitted to Tohoku Medical and Pharmaceutical University Hospital between January 2012 and December 2013. Among the patients, 133 and 147 were positive and negative for PCT, respectively. Patients who were PCT positive were older and more frequently male, had reduced levels of platelets and albumin, and increased levels of aspartate aminotransferase, alanine aminotransferase, blood urea nitrogen, creatinine, and C-reactive protein. Patients who were PCT positive had significantly higher blood culture positivity compared with those who were PCT negative, and the sensitivity and specificity of PCT for detecting positive blood cultures were 74.5% and 59.1%, respectively. Escherichia coli was detected in PCT-positive patients, whereas Staphylococcus epidermidis and Staphylococcus lugdunensis were frequently detected in PCT-negative patients. Levels of PCT were higher in the patients infected with gram-negative rods than those with gram-positive cocci. Furthermore, extended-spectrum β-lactamase (ESBL)-producing bacteria cases showed higher levels of PCT than those of non-ESBL cases. These results suggest that PCT may be a useful biomarker of sepsis, and it might serve as a strong tool to detect patients with severe gram-negative rod bacteremia including ESBL-producing bacteria cases early due to its relative high sensitivity.

  3. Revisiting the Diego Blood Group System in Amerindians: Evidence for Gene-Culture Comigration.

    PubMed

    Bégat, Christophe; Bailly, Pascal; Chiaroni, Jacques; Mazières, Stéphane

    2015-01-01

    Six decades ago the DI*A allele of the Diego blood group system was instrumental in proving Native American populations originated from Siberia. Since then, it has received scant attention. The present study was undertaken to reappraise distribution of the DI*A allele in 144 Native American populations based on current knowledge. Using analysis of variance tests, frequency distribution was studied according to geographical, environmental, and cultural parameters. Frequencies were highest in Amazonian populations. In contrast, DI*A was undetectable in subarctic, Fuegian, Panamanian, Chaco and Yanomama populations. Closer study revealed a correlation that this unequal distribution was correlated with language, suggesting that linguistic divergence was a driving force in the expansion of DI*A among Native Americans. The absence of DI*A in circumpolar Eskimo-Aleut and Na-Dene speakers was consistent with a late migratory event confined to North America. Distribution of DI*A in subtropical areas indicated that gene and culture exchanges were more intense within than between ecozones. Bolstering the utility of classical genetic markers in biological anthropology, the present study of the expansion of Diego blood group genetic polymorphism in Native Americans shows strong evidence of gene-culture comigration.

  4. Revisiting the Diego Blood Group System in Amerindians: Evidence for Gene-Culture Comigration

    PubMed Central

    Bégat, Christophe; Bailly, Pascal; Chiaroni, Jacques; Mazières, Stéphane

    2015-01-01

    Six decades ago the DI*A allele of the Diego blood group system was instrumental in proving Native American populations originated from Siberia. Since then, it has received scant attention. The present study was undertaken to reappraise distribution of the DI*A allele in 144 Native American populations based on current knowledge. Using analysis of variance tests, frequency distribution was studied according to geographical, environmental, and cultural parameters. Frequencies were highest in Amazonian populations. In contrast, DI*A was undetectable in subarctic, Fuegian, Panamanian, Chaco and Yanomama populations. Closer study revealed a correlation that this unequal distribution was correlated with language, suggesting that linguistic divergence was a driving force in the expansion of DI*A among Native Americans. The absence of DI*A in circumpolar Eskimo-Aleut and Na-Dene speakers was consistent with a late migratory event confined to North America. Distribution of DI*A in subtropical areas indicated that gene and culture exchanges were more intense within than between ecozones. Bolstering the utility of classical genetic markers in biological anthropology, the present study of the expansion of Diego blood group genetic polymorphism in Native Americans shows strong evidence of gene-culture comigration. PMID:26148209

  5. Expression of four canine leukocyte adhesion factors in fresh and stored whole blood samples evaluated using a no-lyse, no-wash method.

    PubMed

    Holst, B Ström; Hagberg, M; Lilliehöök, I; Johannisson, A

    2011-02-15

    Expression of four leukocyte adhesion factors on canine leukocytes was studied by flow cytometry using a no-lyse, no-wash method. The effect of fixation and storage for up to 14 days in 1% paraformaldehyde on labelled samples and within assay variation was evaluated. Monoclonal antibodies directed to monocyte marker CD14, and to adhesion molecules CD11a, CD18, CD32 and CD49d were used. Cell surface marker, cell population, time, and the interactions between time and cell marker significantly affected expression of cell adhesion factors. For CD18, there was a significant difference in mean fluorescence intensity (MFI) between fresh and stored samples (P<0.001), but no significant difference between stored samples. The MFIs of CD11a and CD49d were not significantly affected by fixation and storage. The CVs differed significantly depending on cell marker (P<0.001) and cell population (P=0.005). Fixation and storage did not significantly affect the CV. In conclusion, a no-lyse, no-wash method can be applied to canine leukocytes. The effect of fixation and storage on the resulting MFI differs between monoclonal antibodies, and should be evaluated for each antibody before use. The coefficient of variation was generally acceptable, and high CVs were related to a low MFIs or low numbers of analysed cells. Copyright © 2010 Elsevier B.V. All rights reserved.

  6. The Optimization of Molecular Detection of Clinical Isolates of Brucella in Blood Cultures by eryD Transcriptase Gene for Confirmation of Culture-Negative Samples

    PubMed Central

    Tabibnejad, Mahsa; Alikhani, Mohammad Yousef; Arjomandzadegan, Mohammad; Hashemi, Seyed Hamid; Naseri, Zahra

    2016-01-01

    Background Brucellosis is a zoonosis disease which is widespread across the world. Objectives The aim of the present study is the evaluation of culture-negative blood samples. Materials and Methods A total of 100 patients with suspected brucellosis were included in this experimental study and given positive serological tests. Diagnosis was performed on patients with clinical symptoms of the disease, followed by the detection of a titer that was equal to or more than 1:160 (in endemic areas) by the standard tube agglutination method. Blood samples were cultured by a BACTEC 9050 system, and subsequently by Brucella agar. At the same time, DNA from all blood samples was extracted by Qiagen Kit Company (Qia Amp Mini Kit). A molecular assay of blood samples was carried out by detection of eryD transcriptase and bcsp 31 genes in specific double PCR reactions. The specificity of the primers was evaluated by DNA from pure and approved Brucella colonies found in the blood samples, by DNA from other bacteria, and by ordinary PCR. DNA extraction from the pure colonies was carried out by both Qiagen Kit and Chelex 100 methods; the two were compared. Results 39 cases (39%) had positive results when tested by the BACTEC system, and 61 cases (61%) became negative. 23 culture-positive blood samples were randomly selected for PCR reactions; all showed 491 bp for the eryD gene and 223 bp for the bcsp 31 gene. Interestingly, out of 14 culture-negative blood samples, 13 cases showed positive bonds in PCR. The specificity of the PCR method was equal to 100%. DNA extraction from pure cultures was done by both Chelex 100 and Qiagen Kit; these showed the same results for all samples. Conclusions The results prove that the presented double PCR method could be used to detect positive cases from culture-negative blood samples. The Chelex 100 method is simpler and safer than the use of Qiagen Kit for DNA extraction. PMID:27330831

  7. The Optimization of Molecular Detection of Clinical Isolates of Brucella in Blood Cultures by eryD Transcriptase Gene for Confirmation of Culture-Negative Samples.

    PubMed

    Tabibnejad, Mahsa; Alikhani, Mohammad Yousef; Arjomandzadegan, Mohammad; Hashemi, Seyed Hamid; Naseri, Zahra

    2016-04-01

    Brucellosis is a zoonosis disease which is widespread across the world. The aim of the present study is the evaluation of culture-negative blood samples. A total of 100 patients with suspected brucellosis were included in this experimental study and given positive serological tests. Diagnosis was performed on patients with clinical symptoms of the disease, followed by the detection of a titer that was equal to or more than 1:160 (in endemic areas) by the standard tube agglutination method. Blood samples were cultured by a BACTEC 9050 system, and subsequently by Brucella agar. At the same time, DNA from all blood samples was extracted by Qiagen Kit Company (Qia Amp Mini Kit). A molecular assay of blood samples was carried out by detection of eryD transcriptase and bcsp 31 genes in specific double PCR reactions. The specificity of the primers was evaluated by DNA from pure and approved Brucella colonies found in the blood samples, by DNA from other bacteria, and by ordinary PCR. DNA extraction from the pure colonies was carried out by both Qiagen Kit and Chelex 100 methods; the two were compared. 39 cases (39%) had positive results when tested by the BACTEC system, and 61 cases (61%) became negative. 23 culture-positive blood samples were randomly selected for PCR reactions; all showed 491 bp for the eryD gene and 223 bp for the bcsp 31 gene. Interestingly, out of 14 culture-negative blood samples, 13 cases showed positive bonds in PCR. The specificity of the PCR method was equal to 100%. DNA extraction from pure cultures was done by both Chelex 100 and Qiagen Kit; these showed the same results for all samples. The results prove that the presented double PCR method could be used to detect positive cases from culture-negative blood samples. The Chelex 100 method is simpler and safer than the use of Qiagen Kit for DNA extraction.

  8. Reducing time to identification of positive blood cultures with MALDI-TOF MS analysis after a 5-h subculture.

    PubMed

    Verroken, A; Defourny, L; Lechgar, L; Magnette, A; Delmée, M; Glupczynski, Y

    2015-02-01

    Speeding up the turn-around time of positive blood culture identifications is essential in order to optimize the treatment of septic patients. Several sample preparation techniques have been developed allowing direct matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) identification of positive blood cultures. Yet, the hands-on time restrains their routine workflow. In this study, we evaluated an approach whereby MALDI-TOF MS identification without any additional steps was carried out on short subcultured colonies from positive blood bottles with the objective of allowing results reporting on the day of positivity detection. Over a 7-month period in 2012, positive blood cultures detected by 9 am with an automated system were inoculated onto a Columbia blood agar and processed after a 5-h incubation on a MALDI-TOF MicroFlex platform (Bruker Daltonik GmbH). Single-spotted colonies were covered with 1 μl formic acid and 1 μl matrix solution. The results were compared to the validated identification techniques. A total of 925 positive blood culture bottles (representing 470 bacteremic episodes) were included. Concordant identification was obtained in 727 (81.1 %) of the 896 monomicrobial blood cultures, with failure being mostly observed with anaerobes and yeasts. In 17 episodes of polymicrobic bacteremia, the identification of one of the two isolates was achieved in 24/29 (82.7 %) positive cultures. Routine implementation of MALDI-TOF MS identification on young positive blood subcultures provides correct results to the clinician in more than 80 % of the bacteremic episodes and allows access to identification results on the day of blood culture positivity detection, potentially accelerating the implementation of targeted clinical treatments.

  9. Positive blood culture is not associated with increased mortality in patients with sepsis-induced acute respiratory distress syndrome.

    PubMed

    Yang, Szu-Chun; Liao, Kuang-Ming; Chen, Chang-Wen; Lin, Wei-Chieh

    2013-11-01

    Previous studies have demonstrated that positive blood culture could contribute to poorer outcomes in patients with pneumonia. However, the impact of positive blood culture on the outcomes of patients with sepsis-induced acute respiratory distress syndrome (ARDS) has not been evaluated. An observational study that prospectively screened 4861 patients admitted to medical or surgical intensive care units (ICUs) of a tertiary referral centre was performed. Among 4861 admitted patients, 146 diagnosed with sepsis-induced ARDS were enrolled (mean age: 66.1 years). Lower PaO2 /FiO2 , decreased respiratory system compliance, and higher lung injury scores (LIS) on the day of ARDS diagnosis were associated with positive blood cultures (n = 68) rather than negative blood cultures (n = 78). There was no relationship between positive blood culture and in-hospital mortality. Kaplan-Meier estimates also revealed that positive blood culture was not associated with 60-day mortality but with an increased length of stay in the hospital and in the ICU (P = 0.007 and P = 0.016, respectively). Using multivariate logistic regression, higher LIS was independently associated with positive blood culture. In addition, chronic pulmonary disease, lower platelet count, higher LIS, and the development of shock on the diagnosis of ARDS, were independent risk factors for in-hospital mortality. This study suggests that the presence of positive blood culture is not associated with increased mortality; however, the mean durations of hospital and ICU stays in patients with sepsis-induced ARDS are increased. © 2013 The Authors. Respirology © 2013 Asian Pacific Society of Respirology.

  10. Comparison of outcomes between patients with single versus multiple positive blood cultures for Enterococcus: Infection versus illusion?

    PubMed

    Claeys, Kimberly C; Zasowski, Evan J; Lagnf, Abdalhamid M; Rybak, Michael J

    2016-01-01

    Enterococci represent one of the most common causative pathogens of bloodstream infections (BSIs). There is debate in the literature regarding the clinical importance of single versus multiple positive blood cultures for Enterococci. This single-center retrospective study found that patients with multiple positive blood cultures experienced increased inpatient mortality and a shorter median survival. Additionally, BSIs >6.7 days resulted in approximately 20% increased mortality. These results are preliminary and require further exploration.

  11. Fate in humans of the plasticizer, DI (2-ethylhexyl) phthalate, arising from transfusion of platelets stored in vinyl plastic bags. [plasticizer migration into human blood from vinyl plastic bags during transfusion

    NASA Technical Reports Server (NTRS)

    Rubin, R. J.; Schiffer, C. A.

    1975-01-01

    Platelet concentrates were shown to contain 18-38 mg/100 ml of a phthalate plasticizer (DEHP) which arose by migration from the vinyl plastic packs in which the plateletes were prepared and stored. Transfusion of these platelets into 6 adult patients with leukemia resulted in peak blood plasma levels of DEHP ranging from 0.34 - 0.83 mg/100 ml. The blood levels fell mono-exponentially with a mean rate of 2.83 percent per minute and a half-life of 28.0 minutes. Urine was assayed by a method that would measure unchanged DEHP as well as all phthalic acid-containing metabolities. In two patients, at most 60 and 90% of the infused dose, respectively, was excreted in the urine collected for 24 hours post-transfusion. These estimates, however, could be high due to the simultaneous excretion of DEHP remaining from previous transfusions or arising from uncontrolled environmental exposures.

  12. Time course of radiometric detection of positive blood cultures in childhood

    SciTech Connect

    Meadow, W.L.; Schwartz, I.K.

    1986-05-01

    We have determined the time course of radiometric detection of microbial growth in 2348 positive blood culture specimens obtained at Wyler Children's Hospital during a 5-year interval. Overall 72 and 88% of isolates were detected within 48 and 72 hours after sampling, respectively. For pathogenic organisms aerobic detection was generally more rapid and more inclusive than anaerobic detection. At 48 hours of incubation the detection of six potential pathogens (Salmonella sp., Haemophilus influenzae, Group D streptococci, Neisseria meningitidis, coagulase-negative staphylococci, Candida sp.) was significantly delayed compared with detection of other pathogenic organisms recovered from blood. At 72 hours of incubation the detection rates remained less than 95% for H. influenzae, Staphylococcus aureus, Klebsiella sp., coagulase-negative staphylococci, Group D streptococci and Candida sp. These data should assist clinical decisions regarding duration of antibiotic therapy for the presumptive diagnosis of bacteremia in children.

  13. Use of Positive Blood Cultures for Direct Identification and Susceptibility Testing with the Vitek 2 System

    PubMed Central

    de Cueto, Marina; Ceballos, Esther; Martinez-Martinez, Luis; Perea, Evelio J.; Pascual, Alvaro

    2004-01-01

    In order to further decrease the time lapse between initial inoculation of blood culture media and the reporting of results of identification and antimicrobial susceptibility tests for microorganisms causing bacteremia, we performed a prospective study in which specially processed fluid from positive blood culture bottles from Bactec 9240 (Becton Dickinson, Cockeysville, Md.) containing aerobic media were directly inoculated into Vitek 2 system cards (bio-Mérieux, France). Organism identification and susceptibility results were compared with those obtained from cards inoculated with a standardized bacterial suspension obtained following subculture to agar; 100 consecutive positive monomicrobic blood cultures, consisting of 50 gram-negative rods and 50 gram-positive cocci, were included in the study. For gram-negative organisms, 31 of the 50 (62%) showed complete agreement with the standard method for species identification, while none of the 50 gram-positive cocci were correctly identified by the direct method. For gram-negative rods, there were 50% categorical agreements between the direct and standard methods for all drugs tested. The very major error rate was 2.4%, and the major error rate was 0.6%. The overall error rate for gram-negatives was 6.6%. Complete agreement in clinical categories of all antimicrobial agents evaluated was obtained for 19 of 50 (38%) gram-positive cocci evaluated; the overall error rate was 8.4%, with 2.8% minor errors, 2.4% major errors, and 3.2% very major errors. These findings suggest that the Vitek 2 cards inoculated directly from positive Bactec 9240 bottles do not provide acceptable bacterial identification or susceptibility testing in comparison with corresponding cards tested by a standard method. PMID:15297523

  14. PCR evaluation of false-positive signals from two automated blood-culture systems.

    PubMed

    Karahan, Z Ceren; Mumcuoglu, Ipek; Guriz, Haluk; Tamer, Deniz; Balaban, Neriman; Aysev, Derya; Akar, Nejat

    2006-01-01

    Rapid detection of micro-organisms from blood is one of the most critical functions of a diagnostic microbiology laboratory. Automated blood-culture systems reduce the time needed to detect positive cultures, and reduce specimen handling. The false-positive rate of such systems is 1-10%. In this study, the presence of pathogens in 'false-positive' bottles obtained from BACTEC 9050 (Becton Dickinson) and BacT/Alert (Biomérieux) systems was investigated by eubacterial and fungal PCR. A total of 169 subculture-negative aerobic blood-culture bottles (104 BacT/Alert and 65 BACTEC) were evaluated. Both fungal and eubacterial PCRs were negative for all BACTEC bottles. Fungal PCR was also negative for the BacT/Alert system, but 10 bottles (9.6%) gave positive results by eubacterial PCR. Sequence analysis of the positive PCR amplicons indicated the presence of the following bacteria (number of isolates in parentheses): Pasteurella multocida (1), Staphylococcus epidermidis (2), Staphylococcus hominis (1), Micrococcus sp. (1), Streptococcus pneumoniae (1), Corynebacterium spp. (2), Brachibacterium sp. (1) and Arthrobacter/Rothia sp. (1). Antibiotic usage by the patients may be responsible for the inability of the laboratory to grow these bacteria on subcultures. For patients with more than one false-positive bottle, molecular methods can be used to evaluate the microbial DNA in these bottles. False positives from the BACTEC system may be due to elevated patient leukocyte counts or the high sensitivity of the system to background increases in CO(2) concentration.

  15. Transfer of opiorphin through a blood-brain barrier culture model.

    PubMed

    Bocsik, Alexandra; Darula, Zsuzsanna; Tóth, Géza; Deli, Mária A; Wollemann, Mária

    2015-08-01

    Opioid peptides are potent analgesics with therapeutic potential in the treatment of acute and chronic pain. Their efficacy is limited by peptidases (enkephalinases). Opiorphin pentapeptide (QRFSR) is the first characterized human endogenous inhibitor of enkephalinases. The peptide is able to increase the binding and affinity of endogenous opiates to mu opioid receptors; thus, the mechanism of opiorphin may provide a new therapeutic approach in pain management. The analgesic effect of opiorphin was proven in several earlier published in vitro and in vivo studies. Our aim was to test the transfer of opiorphin through a blood-brain barrier model for the first time. The flux of opiorphin was tested on a blood-brain barrier culture model consisting of rat brain endothelial, glial and pericyte cells. Brain endothelial cells in this triple co-culture model form tight monolayers characterized by transendothelial electrical resistance measurement. Relative quantity of the peptide was estimated by mass spectrometry. The transfer of opiorphin through the blood-brain barrier model was estimated to be ∼3%, whereas the permeability coefficient was 0.53 ± 1.36 × 10(-6) cm/s (n = 4). We also observed rapid conversion of N-terminal glutamine into pyroglutamic acid during the transfer experiments. Our results indicate that opiorphin crosses cultured brain endothelial cells in the absence of serum factors in a significant amount. This is in agreement with previous in vivo data showing potentiation of enkephalin-mediated antinociception. We suggest that opiorphin may have a potential as a centrally acting novel drug to treat pain.

  16. Blood

    MedlinePlus

    ... that die or are lost from the body. White Blood Cells White blood cells (WBCs, and also ... of severe pain. previous continue Diseases of the White Blood Cells Neutropenia (pronounced: new-truh-PEE-nee- ...

  17. Characterization of Blood Culture Isolates of Streptococcus dysgalactiae subsp. equisimilis Possessing Lancefield's Group A Antigen

    PubMed Central

    Brandt, Claudia M.; Haase, Gerhard; Schnitzler, Norbert; Zbinden, Reinhard; Lütticken, Rudolf

    1999-01-01

    For three human blood culture isolates of beta-hemolytic streptococci with Lancefield's serogroup A antigen, phylogenetic analysis of the 16S rRNA genes confirmed biochemical identification as Streptococcus dysgalactiae subsp. equisimilis. Genes encoding M or M-like proteins, which are considered to be major virulence determinants in streptococci, were detected in all of these strains. Our data clearly demonstrate that for beta-hemolytic streptococci, the species assignment should not be based on the results of serogrouping alone. PMID:10565964

  18. Direct identification of bacteria in blood culture samples using an electronic nose.

    PubMed

    Trincavelli, Marco; Coradeschi, Silvia; Loutfi, Amy; Söderquist, Bo; Thunberg, Per

    2010-12-01

    In this paper, we introduce a method for identification of bacteria in human blood culture samples using an electronic nose. The method uses features, which capture the static (steady state) and dynamic (transient) properties of the signal from the gas sensor array and proposes a means to ensemble results from consecutive samples. The underlying mechanism for ensembling is based on an estimation of posterior probability, which is extracted from a support vector machine classifier. A large dataset representing ten different bacteria cultures has been used to validate the presented methods. The results detail the performance of the proposed algorithm and show that through ensembling decisions on consecutive samples, significant reliability in classification accuracy can be achieved.

  19. Lack of clinical relevance in routine final subcultures of radiometrically negative BACTEC blood culture vials

    SciTech Connect

    Plorde, J.J.; Carlson, L.G.; Dau, M.E.

    1982-11-01

    During a 38-month period, 10,106 blood specimens were received in the laboratory for culture. These were inoculated into 26,424 vials and processed using the BACTEC radiometric detection system. Of these vials, 1,914 were eventually found to be microbiologically positive. Isolates from 836 vials were judged to be contaminants. In the remaining 1,078 vials, growth was first detected visually or radiometrically in 1,062 and by final subculture in 16. Growth from these sixteen bottles represented 12 clinically significant bacteremic episodes in as many patients. In nine of these episodes, other culture vials from the same patient were positive radiometrically. Therefore, 358 of 361 (99.2%) bacteremic episodes were detected without the benefit of routine final subcultures. The three patients whose bacteremia was missed were diagnosed clinically and placed on appropriate therapy prior to the detection of the bacteremias by final subculture.

  20. IDENTIFICATION OF BACTERIA IN BLOOD CULTURES FROM CLINICALLY ILL CAPTIVE ANTILLEAN MANATEES (TRICHECHUS MANATUS MANATUS).

    PubMed

    Silva, Mariana C O; Attademo, Fernanda F L; Freire, Augusto C B; Sousa, Glaucia P; Luna, Fábia O; Lima, Débora C V; Mota, Rinaldo A; Mendes, Emiko S; Silva, Jean C R

    2017-03-01

    Between September 2001 and March 2013, 62 bacterial cultures (37 aerobic and 25 anaerobic) were performed on 37 blood samples from 23 Antillean manatees ( Trichechus manatus manatus) that were kept in captivity at the Brazilian National Center for Research and Conservation of Aquatic Mammals (CMA) in Pernambuco (CMA-PE) and Alagoas (CMA-AL), Brazil. All of the animals sampled exhibited clinical signs at the time of sampling including abscesses (n = 8), debilitation and anorexia (n = 22), and profound lethargy-moribundity (n = 7). The 4 animals with profound lethargy-moribundity died shortly after sampling of unknown causes. Bacteria were isolated from 15/37 (40.5%) and aerobic blood cultures from 13/23 animals (56.5%). None of the anaerobic cultures were positive. Aeromonas caviae , Aeromonas hydrophila , Aeromonas sp., Escherichia coli , Leclercia adecarboxylata , Pantoea agglomerans , Pseudomonas aeruginosa , Pseudomonas stutzeri , Pseudomonas sp., Sphingomonas paucimobilis , coagulase-negative Staphylococcus, and Staphylococcus epidermidis were each found in only one animal; Staphylococcus spp. was found in two; and Vibrio fluvialis in four. Thirteen samples had only one bacteria isolated, one sample had two bacteria, and one sample had three bacteria isolated. Regarding sex, age group, and origin among the manatees examined, 54.5% (6/11) of the females, 58.3% (7/12) of the males, 40% (2/5) of the calves, 66.7% (8/12) of the juveniles, 50% (3/6) of the adults, 55.5% (10/18) at CMA-PE, and 60% (3/5) at CMA-AL were found to be positive for bacterial growth during at least one sampling time. All Antillean manatees were clinically ill. Regarding clinical signs, bacteria were found in 50% (11/22) of blood samples of the animals showing debilitation and anorexia, 1 of 8 (12.5%) of blood samples of the animals showing abscesses, and 3 of 7 (42.9%) of blood samples of the animals showing profound lethargy-moribundity.

  1. Nurses' competency in drawing blood cultures and educational intervention to reduce the contamination rate.

    PubMed

    Al-Hamad, Arif; Al-Ibrahim, Maha; Alhajhouj, Eman; Al-Alshaikh Jaffer, Waseelah; Altowaileb, Jaffar; Alfaraj, Hassan

    2016-01-01

    Compared with truly negative cultures, false positive blood cultures (BCs) not only increase laboratory work but also prolong the lengths of patient stays, which are likely to increase patient morbidity and costs. The present study aimed to evaluate the effectiveness of a hospital-wide educational intervention on BC contamination rates. Nurses performed all phlebotomies; therefore, educational workshops were offered to all nurses twice a week over a 3-month period. The workshops consisted of a questionnaire, PowerPoint presentation, video show, demonstration of the different materials used to collect BCs, and question session. Data from the questionnaires and laboratory culture results were compared between the 6-month pre- and post-intervention periods. Of the 503 eligible nurses, 216 (42.9%) attended the workshops. The survey identified areas for improvement, which included time of disinfectant application, volume of blood to be cultured, and disinfection of BC bottle tops. Of the 9903 BC sets that were drawn from 3649 patients during the study period, 676 (6.8%) were contaminated. The monthly BC contamination rates for the 6-month pre- and post-intervention periods were 8.1% and 5.2%, respectively, representing a 36% reduction (P=0.008). Only three wards had an acceptable contamination rate of ≤3% before the intervention, compared with eight wards after the intervention. While contamination of BCs can never be completely eliminated, there is evidence that adherence to best practice BC collection techniques can minimize BC contamination, which might be best achieved with a dedicated phlebotomy team.

  2. 3D cultured immortalized human hepatocytes useful to develop drugs for blood-borne HCV

    SciTech Connect

    Aly, Hussein Hassan; Shimotohno, Kunitada; Hijikata, Makoto

    2009-02-06

    Due to the high polymorphism of natural hepatitis C virus (HCV) variants, existing recombinant HCV replication models have failed to be effective in developing effective anti-HCV agents. In the current study, we describe an in vitro system that supports the infection and replication of natural HCV from patient blood using an immortalized primary human hepatocyte cell line cultured in a three-dimensional (3D) culture system. Comparison of the gene expression profile of cells cultured in the 3D system to those cultured in the existing 2D system demonstrated an up-regulation of several genes activated by peroxisome proliferator-activated receptor alpha (PPAR{alpha}) signaling. Furthermore, using PPAR{alpha} agonists and antagonists, we also analyzed the effect of PPAR{alpha} signaling on the modulation of HCV replication using this system. The 3D in vitro system described in this study provides significant insight into the search for novel anti-HCV strategies that are specific to various strains of HCV.

  3. Proportional-Integral-Derivative (PID) Control of Secreted Factors for Blood Stem Cell Culture

    PubMed Central

    Caldwell, Julia; Wang, Weijia; Zandstra, Peter W.

    2015-01-01

    Clinical use of umbilical cord blood has typically been limited by the need to expand hematopoietic stem and progenitor cells (HSPC) ex vivo. This expansion is challenging due to the accumulation of secreted signaling factors in the culture that have a negative regulatory effect on HSPC output. Strategies for global regulation of these factors through dilution have been developed, but do not accommodate the dynamic nature or inherent variability of hematopoietic cell culture. We have developed a mathematical model to simulate the impact of feedback control on in vitro hematopoiesis, and used it to design a proportional-integral-derivative (PID) control algorithm. This algorithm was implemented with a fed-batch bioreactor to regulate the concentrations of secreted factors. Controlling the concentration of a key target factor, TGF-β1, through dilution limited the negative effect it had on HSPCs, and allowed global control of other similarly-produced inhibitory endogenous factors. The PID control algorithm effectively maintained the target soluble factor at the target concentration. We show that feedback controlled dilution is predicted to be a more cost effective dilution strategy compared to other open-loop strategies, and can enhance HSPC expansion in short term culture. This study demonstrates the utility of secreted factor process control strategies to optimize stem cell culture systems, and motivates the development of multi-analyte protein sensors to automate the manufacturing of cell therapies. PMID:26348930

  4. Filter-cultured ARPE-19 cells as outer blood-retinal barrier model.

    PubMed

    Mannermaa, Eliisa; Reinisalo, Mika; Ranta, Veli-Pekka; Vellonen, Kati-Sisko; Kokki, Heidi; Saarikko, Anni; Kaarniranta, Kai; Urtti, Arto

    2010-07-11

    Retinal pigment epithelium (RPE) regulates drug transfer between posterior eye segment and blood circulation, but there is no established RPE cell model for drug delivery studies. We evaluated ARPE-19 filter culture model for this purpose. Passive permeability of 6-carboxyfluorescein, betaxolol and FITC-dextran (40kDa) and active transport of 6-carboxyfluorescein, sodium fluorescein, rhodamine 123, cyclosporine A and digoxin in ARPE-19 model were investigated and compared with isolated bovine RPE-choroid tissue. In addition, barrier properties, and mRNA expression of RPE-specific and melanogenesis-related genes (RPE65, VMD2, CRALBP, OTX-2, MITF-A, TRP-1, tyrosinase) were measured in various culture conditions. The filter grown ARPE-19 cell model showed reasonable barrier properties (TER close to 100Omegacm(2)), but its permeability was slightly higher than that of isolated bovine RPE/choroid specimens. In active transport studies the ARPE-19 model mimics qualitatively the permeability profile of bovine RPE-choroid, but ARPE-19 model underestimates the importance of active transport relative to passive diffusion. Long-term filter-cultured ARPE-19 cells expressed various RPE-specific and melanogenesis-related genes at higher levels than the ARPE-19 cells cultured short-term in flasks. ARPE-19 model can be used to study drug permeation processes in the RPE.

  5. Enhanced Adhesion of Pasteurella multocida to Cultured Turkey Peripheral Blood Monocytes

    PubMed Central

    Pruimboom, Ingrid M.; Rimler, Richard B.; Ackermann, Mark R.

    1999-01-01

    Capsular hyaluronic acid (HA) mediates adhesion of serogroup A strains of Pasteurella multocida to elicited turkey air sac macrophages (TASM). In contrast, freshly isolated turkey peripheral blood monocytes (TPBM) do not bind serogroup A strains. Following culture of TPBM for 6 days in chamber slides, adhesion of the bacteria to TPBM increased gradually. Incubation in chamber slides coated with entactin-collagen IV-laminin (ECL) attachment matrix or exposure to phorbol myristate acetate (PMA) further enhanced the adhesion of P. multocida to TPBM. Addition of HA, but not Arg-Gly-Asp peptide, to TPBM culture inhibited bacterial adherence similarly to the inhibition previously reported for TASM. Exposure of TPBM to monoclonal antibody directed against HA-binding cell surface proteoglycan (CD44) decreased binding of P. multocida. Collectively, these findings indicate that P. multocida adhesion to TPBM is mediated by capsular HA and can be increased by culture on ECL attachment matrix or PMA exposure. Additionally, the findings suggest that the capsular mucopolysaccharide of serogroup A strains of P. multocida recognizes an isoform of CD44 expressed on cultured TPBM. PMID:10024573

  6. Evaluating the role of low-speed centrifugation towards transfecting human peripheral blood mononuclear cell culture.

    PubMed

    Majumdar, M; Ratho, R; Chawla, Y; Singh, M P

    2014-01-01

    The conventional method of transfection of suspension cells by chemical has proven to be very difficult. We present a new transfection protocol, wherein, low-speed centrifugation of cell culture plates immediately after adding the lipid: DNA complex significantly enhances the transfection efficiency. Peripheral blood mononuclear cells (PBMCs) were transfected with BLOCK-iT™ Fluorescent Oligo (scrambled siRNA) and lipofectamine complex using conventional and low-speed centrifugation modified transfection protocols. The efficiency of transfection was determined using flowcytometer and cell viability was checked using MTT assay. Incorporation of low-speed centrifugation significantly enhances the transfection efficiency of BLOCK-iT™ in the suspension culture of PBMCs as compared to conventional transfection method (99.8% vs 28.3%; P < 0.0001), even at a low concentration of 40 picomoles without affecting the cell viability. Centrifugation enhanced transfection (CET) technique is simple, time-saving and novel application without compromising the cell viability in the context of recently popular RNA interference in suspension cultures of PBMCs. This undemanding modification might be applicable to a wide variety of cell lines and solve crucial problem of researchers working with RNA interference in suspension cultures.

  7. A Retrospective Evaluation of Critical Care Blood Culture Yield - Do Support Services Contribute to the "Weekend Effect"?

    PubMed

    Morton, Ben; Nagaraja, Shankara; Collins, Andrea; Pennington, Shaun H; Blakey, John D

    2015-01-01

    The "weekend effect" describes an increase in adverse outcomes for patients admitted at the weekend. Critical care units have moved to higher intensity working patterns to address this with some improved outcomes. However, support services have persisted with traditional working patterns. Blood cultures are an essential diagnostic tool for patients with sepsis but yield is dependent on sampling technique and processing. We therefore used blood culture yield as a surrogate for the quality of support service provision. We hypothesized that blood culture yields would be lower over the weekend as a consequence of reduced support services. We performed a retrospective observational study examining 1575 blood culture samples in a university hospital critical care unit over a one-year period. Patients with positive cultures had, on average, higher APACHE II scores (p = 0.015), longer durations of stay (p = 0.03), required more renal replacement therapy (p<0.001) and had higher mortality (p = 0.024). Blood culture yield decreased with repeated sampling with an increased proportion of contaminants. Blood cultures were 26.7% less likely to be positive if taken at the weekend (p = 0.0402). This effect size is the equivalent to the impact of sampling before and after antibiotic administration. Our study demonstrates that blood culture yield is lower at the weekend. This is likely caused by delays or errors in incubation and processing, reflecting the reduced provision of support services at the weekend. Reorganization of services to address the "weekend effect" should acknowledge the interdependent nature of healthcare service delivery.

  8. Combined education and skin antisepsis intervention for persistently high blood-culture contamination rates in neonatal intensive care.

    PubMed

    O'Connor, C; Philip, R K; Powell, J; Slevin, B; Quinn, C; Power, L; O'Connell, N H; Dunne, C P

    2016-05-01

    Contaminated blood cultures represent challenges regarding diagnosis, duration of hospitalization, antimicrobial use, pharmacy and laboratory costs. Facing problematic neonatal blood culture contamination (3.8%), we instigated a successful intervention combining skin antisepsis using sterile applicators with 2% chlorhexidine gluconate in 70% isopropanol prior to phlebotomy (replacing 70% isopropanol) and staff education. In the six months prior to intervention, 364 neonatal peripheral blood samples were collected. Fourteen (3.8%) were contaminated. In the post-intervention six months, 314 samples were collected. Three (0.96%) were contaminated, representing significant improvement (Fisher's exact test: P = 0.0259). No dermatological sequelae were observed. The improvement has been sustained.

  9. Case series of Bartonella quintana blood culture-negative endocarditis in Washington, DC.

    PubMed

    Ghidey, Fisseha Y; Igbinosa, Osamuyimen; Mills, Kristin; Lai, Leon; Woods, Christian; Ruiz, Maria E; Fishbein, Dawn; Sampath, Rahul; Lowery, Robert; Wortmann, Glenn

    2016-08-01

    Prior studies (predominantly from Europe) have demonstrated blood culture-negative endocarditis due to Bartonella. Our objective was to describe three cases of Bartonella quintana endocarditis identified within one year at a large hospital in Washington, DC, USA. We constructed a descriptive case series from a retrospective review of medical records from April to December 2013 at an 800-bed urban hospital. All three patients (ages: 52, 55 and 57 years) were undomiciled/homeless men with a history of alcoholism. Although they had negative blood cultures, echocardiography demonstrated aortic/mitral valve perforation and regurgitation in one patient, aortic/mitral valve vegetation with mitral regurgitation in the second patient, and aortic valve vegetation with regurgitation in the third patient. The patients had positive Bartonella quintana serum immunoglobulin G (IgG) with negative immunoglobulin M (IgM). PCR on DNA extracted from cardiac valves was positive for Bartonella, and DNA sequencing of PCR amplicons identified Bartonella quintana. Patients received treatment with doxycycline/rifampin or doxycycline/gentamicin. Clinicians should consider Bartonella endocarditis as a differential diagnosis in patients who fit elements of the Duke Criteria, as well as having a history of homelessness and alcoholism.

  10. Shortened Time to Identify Staphylococcus Species from Blood Cultures and Methicillin Resistance Testing Using CHROMAgar

    PubMed Central

    Chihara, Shingo; Hayden, Mary K.; Minogue-Corbett, Eileen; Singh, Kamaljit

    2009-01-01

    The ability to rapidly differentiate coagulase-negative staphylococcus (CoNS) from Staphylococcus aureus and to determine methicillin resistance is important as it affects the decision to treat empiric antibiotic selection. The objective of this study was to evaluate CHROMagar S. aureus and CHROMagar MRSA (Becton Dickinson) for rapid identification of Staphylococcus spp. directly from blood cultures. Consecutive blood culture bottles (BacT Alert 3D SA and SN, bioMérieux) growing gram-positive cocci in clusters were evaluated. An aliquot was plated onto CHROMagar MRSA (C-MRSA) and CHROMagar S. aureus (C-SA) plates, which were read at 12 to 16 hours. C-SA correctly identified 147/147 S. aureus (100% sensitivity); 2 CoNS were misidentified as S. aureus (98% specificity). C-MRSA correctly identified 74/77 MRSA (96% sensitivity). None of the MSSA isolates grew on C-MRSA (100% specificity). In conclusion, CHROMagar is a rapid and sensitive method to distinguish MRSA, MSSA, and coagulase-negative Staphylococcus and may decrease time of reporting positive results. PMID:20016679

  11. Prevalence of extended-spectrum beta-lactamase-producing Enterobacteriaceae isolated from blood cultures in Africa.

    PubMed

    Sangare, S A; Maiga, A I; Guindo, I; Maiga, A; Camara, N; Savadogo, S; Diallo, S; Bougoudogo, F; Armand-Lefevre, L; Andremont, A; Maiga, I I

    2015-09-01

    Extended-spectrum beta-lactamase (ESBL)-producing Enterobacteriaceae have been isolated from many regions of the world. Epidemiological studies are being conducted in Europe, North America, and Asia. No study has however been conducted in Africa to determine the prevalence and distribution of ESBLs on the continent. This literature review aimed at describing the prevalence of ESBL-producing Enterobacteriaceae isolated from blood cultures, as well as the ESBL genes involved at the international level. Our focus was mainly on Africa. We conducted a literature review on PubMed. Articles related to our study field and published between 1996 and 2014 were reviewed and entirely read for most of them, while we only focused on the abstracts of some other articles. Relevant articles to our study were then carefully reviewed and included in the review. The prevalence of ESBL-producing Enterobacteriaceae differs from one country to another. The results of our literature review however indicate that class A ESBLs prevail over the other types. We took into consideration articles focusing on various types of samples to assess the prevalence of ESBL-producing Enterobacteriaceae, but information on isolates from blood cultures is limited. The worldwide prevalence of ESBL-producing Enterobacteriaceae has increased over time. Evidence of ESBL-producing Enterobacteriaceae can be found in all regions of the world. Studies conducted in Africa mainly focused on the Northern and Eastern parts of the continent, while only rare studies were carried out in the rest of the continent.

  12. A Clostridium hathewayi isolate in blood culture of a patient with an acute appendicitis.

    PubMed

    Randazzo, Adrien; Kornreich, Anne; Lissoir, Bénédicte

    2015-10-01

    Clostridium species is a group of anaerobic bacteria constituting the colonic microflora of the intestinal tract. Since molecular methodologies based on 16 rRNA have been established for the classification and the recognition of bacterial species, more than 150 species of Clostridium have been described. Most are considered harmless saprophytes; however, these bacteria may be involved in a wide variety of infections and may be a common cause of enteritis and enterotoxemias in humans. We present the case of a 60-year-old Asian patient admitted in the emergency room with an acute appendicitis where a blood culture showed the presence of a Clostridium hathewayi. This microorganism is an anaerobic bacteria described in 2001 as a Gram negative end-pointed bacillus, usually endospore-forming. It was reclassified in 2014 as Hungatella hathewayi. A literature review has been performed to find articles relating to this bacteria in a clinical case. C. hathewayi is microorganism recently reclassified as Hungatella hathewayi. Its growth in blood cultures has been reported in a few cases in the literature. Although only a few articles have reported its involvement in clinical infections, we assess that its part in the cause of the illness should be evaluated. Copyright © 2015 Elsevier Ltd. All rights reserved.

  13. Vancomycin MIC creep in MRSA blood culture isolates from Germany: a regional problem?

    PubMed

    Kehrmann, J; Kaase, M; Szabados, F; Gatermann, S G; Buer, J; Rath, P-M; Steinmann, J

    2011-05-01

    The aim of this study was to assess the vancomycin MIC distribution for MRSA blood culture isolates over a period of six years in Germany. The study examined 287 MRSA isolates from blood cultures collected at several hospitals in two German cities between 2004 and 2009. The vancomycin MIC was determined by Etest. Genotypic features of the MRSA strains with vancomycin MIC ≥ 1 mg/L were determined by semiautomated repetitive-sequence-based polymerase chain reaction. The range of vancomycin MIC as determined by Etest was 0.25 to 2.0 mg/L. The geometric mean MIC increased by 1.34-fold in city A over the study period (p < 0.05), but there was no meaningful change in city B (a 1.09-fold increase, p > 0.05). Furthermore, in city A a shift in vancomycin MICs occurred as an increase in the percentage of isolates with MIC ≥ 1 mg/L from period one (2004-2006) to period two (2007-2009) (p < 0.0001). Typing results showed that in city A a single clone was predominant (55% of the creep isolates). In this study, the creep phenomenon seems to be a regional problem. We suggest that all hospitals should monitor their local status of elevated vancomycin MICs in invasive MRSA isolates.

  14. Antifungal Activity of Propolis Against Yeasts Isolated From Blood Culture: In Vitro Evaluation.

    PubMed

    Mutlu Sariguzel, Fatma; Berk, Elife; Koc, Ayes Nedret; Sav, Hafize; Demir, Gonca

    2016-09-01

    Due to the failure of available antifungal agents in the treatment of candidemia and the toxic activities of these drugs, a lot of researches are being conducted to develop new nontoxic and effective antifungal agents for optimal control of fungal pathogens. The aim of this study is to evaluate the in vitro antifungal activity of propolis against yeasts isolated from the blood cultures of intensive care unit patients. Seventy-six strains were included in this study. The in vitro antifungal activity of propolis, fluconazole (FLU), and itraconazole (ITR) was investigated by the microdilution broth methods (CLSI guidelines M27-A3 for yeast). The propolis sample was collected from Kayseri, Turkey. Of the 76 isolates, 33 were identified as Candida albicans while 37 were C. parapsilosis, three were C. tropicalis, and three were identified as C. glabrata. The geometric mean range for MIC (μg/ml) with regard to all isolates was 0.077 to 3 μg/ml for FLU and ITR, and 0.375 to 0.70 μg/ml for propolis. It was shown that propolis had significant antifungal activity against all Candida strains and the MIC range of propolis was determined as 0185 to 3 μg/ml. This study demonstrated that propolis had significant antifungal activity against yeasts isolated from blood culture compared with FLU and ITR. The propolis MIC in azole-resistant strains such as C. glabrata was found lower than the FLU MIC. © 2015 Wiley Periodicals, Inc.

  15. Case series of Bartonella quintana blood culture-negative endocarditis in Washington, DC

    PubMed Central

    Ghidey, Fisseha Y.; Mills, Kristin; Lai, Leon; Woods, Christian; Ruiz, Maria E.; Fishbein, Dawn; Sampath, Rahul; Lowery, Robert; Wortmann, Glenn

    2016-01-01

    Introduction: Prior studies (predominantly from Europe) have demonstrated blood culture-negative endocarditis due to Bartonella. Our objective was to describe three cases of Bartonella quintana endocarditis identified within one year at a large hospital in Washington, DC, USA. Case presentation: We constructed a descriptive case series from a retrospective review of medical records from April to December 2013 at an 800-bed urban hospital. All three patients (ages: 52, 55 and 57 years) were undomiciled/homeless men with a history of alcoholism. Although they had negative blood cultures, echocardiography demonstrated aortic/mitral valve perforation and regurgitation in one patient, aortic/mitral valve vegetation with mitral regurgitation in the second patient, and aortic valve vegetation with regurgitation in the third patient. The patients had positive Bartonella quintana serum immunoglobulin G (IgG) with negative immunoglobulin M (IgM). PCR on DNA extracted from cardiac valves was positive for Bartonella, and DNA sequencing of PCR amplicons identified Bartonella quintana. Patients received treatment with doxycycline/rifampin or doxycycline/gentamicin. Conclusion: Clinicians should consider Bartonella endocarditis as a differential diagnosis in patients who fit elements of the Duke Criteria, as well as having a history of homelessness and alcoholism. PMID:28348772

  16. Rapid Identification of Microorganisms by FilmArray Blood Culture Identification Panel Improves Clinical Management in Children.

    PubMed

    Ray, Stephen T J; Drew, Richard J; Hardiman, Fiona; Pizer, Barry; Riordan, Andrew

    2016-05-01

    Blood cultures are a common investigation for children admitted to hospital. In routine practice, it takes at least 24 hours to identify an organism as a contaminant or clinically significant. FilmArray Blood Culture Identification Panel (FA-BCIP) is a multiplex polymerase chain reaction that can detect 24 pathogens within 1 hour. We assessed whether results from FA-BCIP lead to changes in clinical management in a tertiary referral paediatric hospital. We prospectively studied children having blood cultures taken at our tertiary children's hospital. Blood cultures were monitored and organisms identified using standard methods. FA-BCIP was performed when growth was initially detected in first positive blood cultures per episode, between January 1 and June 30, 2014. Assessment of whether the FA-BCIP result altered clinical management was made, specifically focused on antimicrobial stewardship and length of stay. FA-BCIP was done on 117 positive blood cultures; 74 (63%) grew clinically significant organisms, 43 (37%) grew contaminants. FA-BCIP results were judged to alter clinical management in 63 of the 117 episodes (54%). Antimicrobials were started/altered in 23 (19%) episodes and de-escalated/withheld/stopped in 29 (25%) episodes. Ten children were discharged from hospital earlier, which saved a cumulative total of 14 bed days. Rapid identification of microorganisms in pediatric blood cultures by FA-BCIP, led to changes in clinical management for half of the episodes. This improved antimicrobial stewardship and allowed early discharge from hospital for 10% of children. Future studies should focus on how best to use this technology in a cost-effective manner.

  17. NanoFlares for the detection, isolation, and culture of live tumor cells from human blood

    PubMed Central

    Halo, Tiffany L.; McMahon, Kaylin M.; Angeloni, Nicholas L.; Xu, Yilin; Wang, Wei; Chinen, Alyssa B.; Malin, Dmitry; Strekalova, Elena; Cryns, Vincent L.; Cheng, Chonghui; Mirkin, Chad A.; Thaxton, C. Shad

    2014-01-01

    Metastasis portends a poor prognosis for cancer patients. Primary tumor cells disseminate through the bloodstream before the appearance of detectable metastatic lesions. The analysis o