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Sample records for donor immunoassay cedia

  1. Cross-reactivity of the CEDIA buprenorphine assay in drugs-of-abuse screening: influence of dose and metabolites of opioids

    PubMed Central

    Berg, Jon Andsnes; Schj鴗t, Jan; Fossan, Kjell O; Riedel, Bettina

    2015-01-01

    Purpose The cloned enzyme donor immunoassay (CEDIA) for buprenorphine is applied for both urine drugs-of-abuse screening and compliance monitoring. Sensitivity, specificity, and optimal cutoff of this assay have differed between studies. This may indicate that cross-reactivity has to be taken into account during assay evaluation. We therefore investigated the performance of the CEDIA buprenorphine assay for use in our patient population and explored the impact of cross-reactivity on assay accuracy. Methods The CEDIA buprenorphine assay and high-performance liquid chromatography杢andem mass spectrometry were employed to analyze drugs-of-abuse in urine samples from a healthy drug-na飗e male volunteer after intake of two tablets of a prescription drug containing 400 mg paracetamol +30 mg codeine phosphate, and in urine samples (n=2,272) from drug-addicted patients. Receiver operating characteristic analyses were performed to express the diagnostic accuracy of the CEDIA buprenorphine assay. Results CEDIA buprenorphine was positive in one urine sample from the drug-na飗e person after intake of the prescription drug. Twenty-five (1.1%) of the patient urine samples were positive for buprenorphine by CEDIA, but negative by high-performance liquid chromatography杢andem mass spectrometry. Codeine, morphine, and their respective metabolites were prevalent in samples that were false positive for buprenorphine. The specificity of the CEDIA buprenorphine assay increased to 99.7% when the cutoff was increased from 5 ng/mL to 10 ng/mL. Conclusion Intake of a therapeutic dose of codeine can yield a false-positive CEDIA buprenorphine result. Additive effects from metabolites of codeine contribute to cross-reactivity in concentrations much lower than listed in the manufacturer抯 cross-reactivity guide. Raising the cutoff from 5 ng/mL to 10 ng/mL increased the diagnostic accuracy. Clinicians should be informed about the risk of false-positive results with the CEDIA buprenorphine assay. PMID:26604854

  2. An ImmunoChip prototype for simultaneous detection of antiepileptic drugs using an enhanced one-step homogeneous immunoassay1񁺑4

    PubMed Central

    Yang, Xiaoyun; Janatova, Jarmila; Juenke, JoEtta M.; McMillin, Gwendolyn A.; Andrade, Joseph D.

    2007-01-01

    The development and characterization of a one-step homogeneous immunoassay-based multi-well ImmunoChip is reported for the simultaneous detection and quantitation of antiepileptic drugs (AEDs). The assay platform utilizes a Cloned Enzyme Donor Immunoassay (CEDIA), and a Beta-Glo assay system for generation of bioluminescent signal. Results of the one-step CEDIA for three AEDs (CBZ, carbamazepine; PHT, phenytoin; VPA, valproic acid), in the presence of serum, correlate well with the values determined by Fluorescence Polarization Immunoassay. CEDIA intra-assay and inter-assay coefficients of variation are lower than 10%. A microfabrication process, xurography, was utilized to produce the multi-well ImmunoChip. Assay reagents were dispensed, and lyophilized, in a three-layer pattern. The multi-well ImmunoChip prototype was used to detect and quantify AEDs in serum samples containing all three drugs. Luminescent signals generated from each well were recorded with a Charged Coupled Device (CCD) camera. The assays performed on an ImmunoChip were fast (5 min), requiring only small volumes of both the reagents (< 1 ?l per well) and the serum sample. The ImmunoChip assay platform described herein may be well suited for therapeutic monitoring of drugs and metabolites at the point-of-care. PMID:17448436

  3. Immunoassay

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Immunoassays are analytical methods that employ antibodies or molecules derived from antibodies for the essential binding reactions. The choice of immunoassay system for food safety analysis depends on the analyte, the matrix, and the requirements of the analysis (speed, throughput, sensitivity, spe...

  4. Immunoassays

    NASA Astrophysics Data System (ADS)

    Hsieh, Y.-H. Peggy

    Immunochemistry is a relatively new science that has developed rapidly in the last few decades. One of the most useful analytical developments associated with this new science is immunoassay. Originally immunoassays were developed in medical settings to facilitate the study of immunology, particularly the antibody-antigen interaction. Immunoassays now are finding widespread applications outside the clinical field because they are appropriate for a wide range of analytes ranging from proteins to small organic molecules. In the food analysis area, immunoassays are widely used for chemical residue analysis, identification of bacteria and viruses, and detection of proteins in food and agricultural products. Protein detection is important for determination of allergens and meat species content, seafood species identification, and detection of genetically modified plant tissues. While immunoassays of all formats are too numerous to cover completely in this chapter, there are several procedures that have become standard for food analysis because of their specificity, sensitivity, and simplicity.

  5. Time-Resolved Analysis of a Highly Sensitive F鰎ster Resonance Energy Transfer Immunoassay Using Terbium Complexes as Donors and Quantum Dots as Acceptors

    PubMed Central

    Hildebrandt, Niko; Charbonni鑢e, Lo颿 J.; L鰄mannsr鯾en, Hans-Gerd

    2007-01-01

    CdSe/ZnS core/shell quantum dots (QDs) are used as efficient F鰎ster Resonance Energy Transfer (FRET) acceptors in a time-resolved immunoassays with Tb complexes as donors providing a long-lived luminescence decay. A detailed decay time analysis of the FRET process is presented. QD FRET sensitization is evidenced by a more than 1000-fold increase of the QD luminescence decay time reaching ca. 0.5 milliseconds, the same value to which the Tb donor decay time is quenched due to FRET to the QD acceptors. The FRET system has an extremely large F鰎ster radius of approx. 100 and more than 70% FRET efficiency with a mean donor-acceptor distance of ca. 84 , confirming the applied biotin-streptavidin binding system. Time-resolved measurement allows for suppression of short-lived emission due to background fluorescence and directly excited QDs. By this means a detection limit of 18 attomol QDs within the immunoassay is accomplished, an improvement of more than two orders of magnitude compared to commercial systems. PMID:18273412

  6. In Situ Generation of Electron Donor to Assist Signal Amplification on Porphyrin-Sensitized Titanium Dioxide Nanostructures for Ultrasensitive Photoelectrochemical Immunoassay.

    PubMed

    Shu, Jian; Qiu, Zhenli; Zhuang, Junyang; Xu, Mingdi; Tang, Dianping

    2015-10-28

    An ultrasensitive photoelectrochemical (PEC) immunoassay protocol for quantitative detection of low-abundant proteins at a low potential was designed by utilizing porphyrin-sensitized titanium dioxide (TiO2) nanostructures. Experimental results demonstrated that the water-soluble 5,10,15,20-tetra(4-sulfophenyl)-21H,23H-porphyrin (TSPP) could be bound onto titanium dioxide via the sulfonic group. TSPP-sensitized TiO2 nanostructures exhibited better photoelectrochemical responses and stability in comparison with TiO2 nanoparticles alone under continuous illumination. Using carcinoembryonic antigen (CEA) as a model analyte, a typical PEC immunosensor by using TSPP-TiO2 as the affinity support of anti-CEA capture antibody (Ab1) to facilitate the improvement of photocurrent response was developed. Bioconjugates of secondary antibody and glucose oxidase with gold nanoparticles (Ab2/GOx-AuNPs) was introduced by an antigen-antibody immunoreaction. AuNP acted as a powerful scaffold to bind with bioactive molecules, while GOx catalyzed glucose to in situ generate hydrogen peroxide (H2O2). The generated H2O2 as a sacrificial electron donor could be oxidized by the photogenerated holes to assist the signal amplification at a low potential under light excitation, thus eliminating interference from other species coexisting in the samples. Under optimal conditions, the PEC immunosensor showed a good linear relationship ranging from 0.02 to 40 ng mL(-1) with a low detection limit of 6 pg mL(-1) CEA. The precision, reproducibility, and specificity were acceptable. In addition, the method accuracy was also evaluated for quantitatively monitoring human serum samples, giving results matching with the referenced CEA ELISA kit. PMID:26451956

  7. Immunoassay techniques.

    PubMed

    Wheeler, Michael J

    2013-01-01

    No other development has had such a major impact on the measurement of hormones as immunoassay. Reagents and assay kits can now be bought commercially but not for the more esoteric or new hormones. This chapter explains the basics of the immunoassay reaction and gives simple methods for immunoassays and immunometric assays and for the production of reagents for both antigenic and hapten hormones. Alternative methods are given for the preparation of labeled hormones as well as several possible separation procedures. The methods described here have been previously used in a wide range of assays and have stood the test of time. They will allow the production of usable immunoassays in a relatively short period of time. PMID:23996355

  8. Colloidal nanomaterial-based immunoassay.

    PubMed

    Teste, Bruno; Descroix, Stephanie

    2012-06-01

    Nanomaterials have been widely developed for their use in nanomedicine, especially for immunoassay-based diagnosis. In this review we focus on the use of nanomaterials as a nanoplatform for colloidal immunoassays. While conventional heterogeneous immunoassays suffer from mass transfer limitations and consequently long assay time, colloidal immunosupports allow target capture in the entire volume, thus speeding up reaction kinetics and shortening assay time. Owing to their wide range of chemical and physical properties, nanomaterials are an interesting candidate for immunoassay development. The most popular colloidal nanomaterials for colloidal immunoassays will be discussed, as well as their influence on immune reactions. Recent advances in nanomaterial applications for different formats of immunoassays will be reported, such as nanomaterial-based indirect immunoassays, optical-based agglutination immunoassays, resonance energy transfer-based immunoassays and magnetic relaxation-based immunoassays. Finally, the future of using nanomaterials for homogeneous immunoassays dedicated to clinical diagnosis will be discussed. PMID:22734642

  9. Immunoassays in Biotechnology

    EPA Science Inventory

    Immunoassays have broad applications for a wide variety of important biological compounds and environmental contaminants. Immunoassays can detect the presence of an antigen in the human body, a pollutant in the environment, or a critical antibody in a patient抯 serum to develop a...

  10. PCB immunoassay performance evaluation

    SciTech Connect

    Schreiber, J.A.; Pedersen, T.A.

    1995-12-31

    Field analytical technologies are becoming increasing relied upon in the performance of site investigations and remediation verification activities. Immunoassay screening methods provide a means to perform rapid analyses of soil, groundwater and waste samples in the field. When used in conjunction with a dynamic adaptive sampling/analyses approach immunoassay field screening data provides valuable information for decision making. Soil samples that were collected as part of site characterization and cleanup verification studies at an industrial facility in the northeast were analyzed using immunoassay kits to allow for expedited decisions on investigation and remediation approaches. The 161 samples collected were analyzed in the field using the Millipore EnviroGard{trademark} PCB and EnSys PCB RISc{trademark} immunoassay test methods. Confirmation analyses on all the samples were conducted in a laboratory using EPA Method 8080. Correlation of the field and laboratory results indicate that false negative and false positive rates vary depending on contaminant concentration and sample the matrix properties. Non-parametric statistical methods used to compare the quantified laboratory data to the immunoassay test range results show no significant difference at the 95 percent confidence intervals for some of the data sets although false positive and negative error in excess of 10 percent were observed in some cases. The results of the evaluation suggest that control of error rates can best be achieved by ensuring rigorous control of the field methodologies.

  11. Hydrogel nanoparticle based immunoassay

    DOEpatents

    Liotta, Lance A; Luchini, Alessandra; Petricoin, Emanuel F; Espina, Virginia

    2015-04-21

    An immunoassay device incorporating porous polymeric capture nanoparticles within either the sample collection vessel or pre-impregnated into a porous substratum within fluid flow path of the analytical device is presented. This incorporation of capture particles within the immunoassay device improves sensitivity while removing the requirement for pre-processing of samples prior to loading the immunoassay device. A preferred embodiment is coreshell bait containing capture nanoparticles which perform three functions in one step, in solution: a) molecular size sieving, b) target analyte sequestration and concentration, and c) protection from degradation. The polymeric matrix of the capture particles may be made of co-polymeric materials having a structural monomer and an affinity monomer, the affinity monomer having properties that attract the analyte to the capture particle. This device is useful for point of care diagnostic assays for biomedical applications and as field deployable assays for environmental, pathogen and chemical or biological threat identification.

  12. Updates in immunoassays: bacteriology.

    PubMed

    Josko, Deborah

    2012-01-01

    There are many immunoassays available that provide rapid, accurate and sensitive results. The intent of this article was to provide a brief overview of some of the products and methodologies available for clinical use and to discuss some of the principles behind the methodology and instrumentation. In the area of infectious disease, the use of immunoassays ensures rapid turnaround times that will result in the administration of prompt, accurate treatment for the patient. Ultimately, this will improve overall patient outcomes while possibly decreasing the costs associated with increased hospital stay. In conclusion, immunoassays are essentially easy to perform, cost-effective, produce highly sensitive and specific results, and allow the medical laboratory professional the ability to report accurate results in a timely manner. PMID:22953518

  13. [Electro chemiluminescence immunoassay].

    PubMed

    Sano, M; Tatsumi, N

    1996-11-01

    Boehringer Mannheim is starting into a new age of immunoassay systems. A new immunoassay technology should allow the development of random access immunoassay analyzers with short incubation times. To satisfy the such matter, we have found a new technology ECL (Electro ChemiLuminescence). This technology has been licensed to Boehringer Mannheim by IGEN inc. (Maryland, USA). It's based on a microparticle solid phase coated streptavidin and a very promising complete new electro chemiluminescence technology. The ruthenium complex label used in the system will enables extremely stable reagents compared to enzyme conjugates. The test principle is the following : 1) pipette a sample, biotinylated antibodies, antibodies labeled ruthenium complex, and streptavidin coated microparticles into a reaction cup, 2) after incubation, the reaction mixture is aspirated into an electrochemical measuring cell, and unbound conjugate are washed away by tripropylamine (TPA) and a magnet, 3) by applying a potential to electrode, the ruthenium complex become to exciting stage, and the signal generation initiates. The analyzers using this technology have developed 2 type of fully automated instrument. The use of this new technology and these analyzers will make immunoassay analyses as convenient as clinical chemistry. PMID:8953939

  14. IMMUNOASSAY HUMAN EXPOSURE STUDIES

    EPA Science Inventory

    The Human Exposure Research Branch has developed several enzyme-linked immunosorbent assay (ELISA) methods to support human exposure assessment studies. Immunoassays to detect low levels (<10 ng/mL) of chlorpyrifos in food, track-in dirt and house dust have been applied to sam...

  15. Mass spectrometric immunoassay

    DOEpatents

    Nelson, Randall W (Phoenix, AZ); Williams, Peter (Phoenix, AZ); Krone, Jennifer Reeve (Granbury, TX)

    2007-12-04

    Rapid mass spectrometric immunoassay methods for detecting and/or quantifying antibody and antigen analytes utilizing affinity capture to isolate the analytes and internal reference species (for quantification) followed by mass spectrometric analysis of the isolated analyte/internal reference species. Quantification is obtained by normalizing and calibrating obtained mass spectrum against the mass spectrum obtained for an antibody/antigen of known concentration.

  16. Mass spectrometric immunoassay

    DOEpatents

    Nelson, Randall W.; Williams, Peter; Krone, Jennifer Reeve

    2005-12-13

    Rapid mass spectrometric immunoassay methods for detecting and/or quantifying antibody and antigen analytes utilizing affinity capture to isolate the analytes and internal reference species (for quantification) followed by mass spectrometric analysis of the isolated analyte/internal reference species. Quantification is obtained by normalizing and calibrating obtained mass spectrum against the mass spectrum obtained for an antibody/antigen of known concentration.

  17. Mass spectrometric immunoassay

    DOEpatents

    Nelson, Randall W; Williams, Peter; Krone, Jennifer Reeve

    2013-07-16

    Rapid mass spectrometric immunoassay methods for detecting and/or quantifying antibody and antigen analytes utilizing affinity capture to isolate the analytes and internal reference species (for quantification) followed by mass spectrometric analysis of the isolated analyte/internal reference species. Quantification is obtained by normalizing and calibrating obtained mass spectrum against the mass spectrum obtained for an antibody/antigen of known concentration.

  18. Universal phosphorescence immunoassay.

    PubMed

    Mantrova EYu; Demcheva, M V; Savitsky, A P

    1994-05-15

    The aim of this study is to develop a universal phosphorescence immunoassay method using monoclonal antibodies to Pd-coproporphyrin (Pd-CP) and conjugates of various proteins with Pd-CP. Pd-CP and monoclonal antibodies obtained allow a convenient method for the determination of various antigens to be developed. The conditions for immunological reactions with Pd-CP were optimized with respect to the components affecting the nonspecific binding Pd-CP and Pd-CP conjugates. PMID:8059936

  19. Mass spectrometric immunoassay

    SciTech Connect

    Nelson, R.W.; Krone, J.R.; Bieber, A.L.; Williams, P.

    1995-04-01

    A new, general method of immunoassay is demonstrated. The approach is based on the microscale immunoaffinity capture of target antigens followed by mass-specific identification and quantitation using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry. Immunoaffinity capture of antigens effectively overcomes signal suppression effects typically encountered during traditional matrix-assisted laser desorption/ionization analysis of complex biological mixtures while simultaneously concentrating the analyte into a small volume. Sample incubation and processing methods were such that a typical analysis could be performed in less than 1 h while subnanomolar sensitivities were maintained. The technique has been used for the rapid, selective, and quantitative screening of human blood for the presence of myotoxin a, and Mojave toxin from the venoms of the prairie rattlesnake, Crotalus virdis virdis, and the Mojave rattlesnake, Crotalus scutulatus scutulatus. 18 refs., 8 figs.

  20. Morphological resonances for multicomponent immunoassays

    NASA Astrophysics Data System (ADS)

    Whitten, W. B.; Shapiro, M. J.; Ramsey, J. M.; Bronk, B. V.

    1995-06-01

    An immunoassay technique capable of detecting and identifying a number of species of microorganisms in a single analysis is described. The method uses optical-resonance size discrimination of microspheres to identify antibodies to which stained microorganisms are bound.

  1. Protein Adsorption in Microengraving Immunoassays

    PubMed Central

    Song, Qing

    2015-01-01

    Microengraving is a novel immunoassay forcharacterizing multiple protein secretions from single cells. During the immunoassay, characteristic diffusion and kinetic time scales ?D and ?K determine the time for molecular diffusion of proteins secreted from the activated single lymphocytes and subsequent binding onto the glass slide surface respectively. Our results demonstrate that molecular diffusion plays important roles in the early stage of protein adsorption dynamics which shifts to a kinetic controlled mechanism in the later stage. Similar dynamic pathways are observed for protein adsorption with significantly fast rates and rapid shifts in transport mechanisms when C0* is increased a hundred times from 0.313 to 31.3. Theoretical adsorption isotherms follow the trend of experimentally obtained data. Adsorption isotherms indicate that amount of proteins secreted from individual cells and subsequently captured on a clean glass slide surface increases monotonically with time. Our study directly validates that protein secretion rates can be quantified by the microengraving immunoassay. This will enable us to apply microengraving immunoassays to quantify secretion rates from 104105 single cells in parallel, screen antigen-specific cells with the highest secretion rate for clonal expansion and quantitatively reveal cellular heterogeneity within a small cell sample. PMID:26501282

  2. Bioelectrochemical Immunoassay of Polychlorinated Biphenyl

    SciTech Connect

    Lin, Ying-Ying; Liu, Guodong; Wai, Chien M.; Lin, Yuehe

    2008-04-01

    A simple, rapid, and highly sensitive bioelectrochemical immunoassay method based on magnetic beads (MBs) and disposable screen-printed electrodes (SPE) has been developed to detect polychlorinated biphenyls (PCBs). The principle of this bioassay is based on a direct competitive enzyme-linked immunosorbent assay using PCB-antibody-coated MBs and horseradish peroxidase (HRP)-labeled PCB (HRP-PCB). A magnetic process platform was used to mix and shake the samples during the immunoreactions and to separate free and unbound reagents after the liquid-phase competitive immunoreactions among PCB-antibody-coated MBs, PCB analyte, and HRP-PCB. After a complete immunoassay, the HRP tracers attached to MBs were transferred to a substrate solution containing o-aminophenol and hydrogen peroxide for electrochemical detection. The different parameters, including the amount of HRP-PCB conjugates, immunoreaction time, and the concentration of substrate that governs the analytical performance of the immunoassay have been studied in detail and optimized. The detection limit of 5 pg mL-1 was obtained under optimum experimental conditions. The performance of this bioelectrochemical immunoassay was successfully evaluated with untreated river water spiked with PCBs, and the results were validated by commercial PCB enzyme-linked immunosorbent assay kit, indicating that this convenient and sensitive technique offers great promise for decentralized environmental application and trace PCBs monitoring.

  3. Competitive Homogeneous Immunoassay for Rapid Serodiagnosis of Hantavirus Disease.

    PubMed

    Hepojoki, Satu; Rusanen, Juuso; Hepojoki, Jussi; Nurmi, Visa; Vaheri, Antti; Lundkvist, 舓e; Hedman, Klaus; Vapalahti, Olli

    2015-07-01

    In this study, we describe a competitive homogeneous immunoassay that makes use of F鰎ster resonance energy transfer (FRET) in rapid detection of pathogen-specific antibodies. The assay principle is based on competition between a monoclonal antibody (MAb) and serum antibodies to a given antigen. In the assay, named competitive FRET immunoassay (CFRET-IA), the FRET signal is induced if MAb carrying a donor label binds to an acceptor-labeled antigen. Specific antibodies in serum compete for antigen binding, resulting in reduced FRET signal. The proof-of-principle for the assay was obtained using donor-labeled Puumala virus nucleocapsid protein (PUUV-N) and acceptor-labeled anti-PUUV-N MAb. The assay was evaluated by analyzing 329 clinical samples comprising 101 from individuals with acute PUUV infection, 42 from individuals with past infection, and 186 from individuals with PUUV-seronegative sera, and the results were compared to those of reference tests. The rapid serodiagnostic test we introduced herein performed with 100% sensitivity and 99% specificity for diagnosing acute hantavirus disease. PMID:25972427

  4. Competitive Homogeneous Immunoassay for Rapid Serodiagnosis of Hantavirus Disease

    PubMed Central

    Rusanen, Juuso; Hepojoki, Jussi; Nurmi, Visa; Vaheri, Antti; Lundkvist, 舓e; Hedman, Klaus; Vapalahti, Olli

    2015-01-01

    In this study, we describe a competitive homogeneous immunoassay that makes use of F鰎ster resonance energy transfer (FRET) in rapid detection of pathogen-specific antibodies. The assay principle is based on competition between a monoclonal antibody (MAb) and serum antibodies to a given antigen. In the assay, named competitive FRET immunoassay (CFRET-IA), the FRET signal is induced if MAb carrying a donor label binds to an acceptor-labeled antigen. Specific antibodies in serum compete for antigen binding, resulting in reduced FRET signal. The proof-of-principle for the assay was obtained using donor-labeled Puumala virus nucleocapsid protein (PUUV-N) and acceptor-labeled anti-PUUV-N MAb. The assay was evaluated by analyzing 329 clinical samples comprising 101 from individuals with acute PUUV infection, 42 from individuals with past infection, and 186 from individuals with PUUV-seronegative sera, and the results were compared to those of reference tests. The rapid serodiagnostic test we introduced herein performed with 100% sensitivity and 99% specificity for diagnosing acute hantavirus disease. PMID:25972427

  5. Scheme for the selection of measurement uncertainty models in blood establishments' screening immunoassays.

    PubMed

    Pereira, Paulo; Westgard, James O; Encarna玢o, Pedro; Seghatchian, Jerard; de Sousa, Gracinda

    2015-02-01

    Blood establishments routinely perform screening immunoassays to assess safety of the blood components. As with any other screening test, results have an inherent uncertainty. In blood establishments the major concern is the chance of false negatives, due to its possible impact on patients' health. This article briefly reviews GUM and diagnostic accuracy models for screening immunoassays, recommending a scheme to support the screening laboratories' staffs on the selection of a model considering the intended use of the screening results (i.e., post-transfusion safety). The discussion is grounded on a "risk-based thinking", risk being considered from the blood donor selection to the screening immunoassays. A combination of GUM and diagnostic accuracy models to evaluate measurement uncertainty in blood establishments is recommended. PMID:25620757

  6. Isotope-labeled immunoassays without radiation waste

    E-print Network

    Hammock, Bruce D.

    Isotope-labeled immunoassays without radiation waste Guomin Shan*, Wei Huang*, Shirley J. Gee with radioactive materials, and (iii) short shelf-life of the labeled re- agents. The advantage of isotopic with ELISA or fluorescent detection systems. We developed a format for isotope label immunoassay

  7. Survey of immunoassay techniques for biological analysis

    SciTech Connect

    Burtis, C.A.

    1986-10-01

    Immunoassay is a very specific, sensitive, and widely applicable analytical technique. Recent advances in genetic engineering have led to the development of monoclonal antibodies which further improves the specificity of immunoassays. Originally, radioisotopes were used to label the antigens and antibodies used in immunoassays. However, in the last decade, numerous types of immunoassays have been developed which utilize enzymes and fluorescent dyes as labels. Given the technical, safety, health, and disposal problems associated with using radioisotopes, immunoassays that utilize the enzyme and fluorescent labels are rapidly replacing those using radioisotope labels. These newer techniques are as sensitive, are easily automated, have stable reagents, and do not have a disposal problem. 6 refs., 1 fig., 2 tabs.

  8. Competitive Immunoassays Using Antigen Microarrays.

    PubMed

    Zhang, Zhaowei; Hu, Weihua; Zhang, Qi; Li, Peiwu; Li, Changming

    2016-01-01

    In this work, a non-fouling antigen competitive immunoassay microarray based on the polymer brush is reported to detect multiple mycotoxins. The detection is achieved by utilizing highly specific monoclonal antibodies produced in our laboratory. The polymer brush, poly[oligo(ethylene glycol) methacrylate-co-glycidyl methacrylate] (POEGMA-co-GMA), is synthesized via surface-initiated atom transfer radical polymerization (SI-ATRP) on standard glass slides. In the polymer brush, the epoxy groups of glycidyl methacrylate (GMA) residues provide covalent binding sites for spotted antigens. Moreover, the abundant poly(ethylene glycol) (PEG) side chains in the brush are able to ultimately suppress the nonspecific protein adsorption in solution (non-fouling). The polymer brush shows a high and uniform protein loading, along with a high resistance to nonspecific protein absorption that are both important to achieve a highly sensitive immunoassay. As a demonstration of a multiplex assay, aflatoxin B1 (AFB1), ochratoxin A (OTA), and zearalenone (ZEN) are selected as antigen targets for simultaneous detections using the microarray. PMID:26614080

  9. Automated immunoassays for 25-hydroxyvitamin D (25-OHD): do plasticisers interfere?

    PubMed

    Carter, G D; Jones, J; Ketheeswaran, M; Shannon, J; Singh, B; Kearney, E; Berry, J L

    2015-04-01

    The international quality assessment scheme for vitamin D metabolites (DEQAS) was established in 1989. The scheme involves the quarterly distribution of 5 serum samples prepared from blood collected in plain plastic bags. Following transfer of the donors to a clinic using different bags, sera were found to contain a contaminant that interfered in both the local LC-MS/MS assay and the NIST reference measurement procedure for 25-OHD. It seemed likely that the contaminant was a substance, possibly a plasticiser, leached from the plastic bag. It was subsequently suggested that the unidentified contaminant might also cause interference in certain automated non-extraction assays for 25-OHD. This was investigated in 3 automated immunoassays by comparing serum 25-OHD results from blood collected simultaneously into plain glass tubes and plastic bags. There was no significant difference in results, indicating that the leached substance had no effect on any of the 3 immunoassays examined. PMID:25448742

  10. Finding a Donor

    MedlinePLUS

    ... hope for somebody else A mother's plea - a child's fight Your self-sacrifice saved our daughters life A single mother's transplant story Knowledge is power Donor stories Paul, marrow donor, explains donation process ...

  11. GIFT AGREEMENT BETWEEN NAME OF DONOR(S)

    E-print Network

    Thompson, Michael

    1 GIFT AGREEMENT BETWEEN NAME OF DONOR(S) ("the Donor(s)") AND McMASTER UNIVERSITY ("the University") I. THE GIFT (NAME OF DONOR(S)) has generously made a Gift of (GIFT/PLEDGE $) to McMaster University (hereinafter the "Gift"). This Gift/Pledge will be provided in the form of (METHOD OF PAYMENT) according

  12. GIFT AGREEMENT BETWEEN (NAME OF DONOR(S))

    E-print Network

    Thompson, Michael

    1 GIFT AGREEMENT BETWEEN (NAME OF DONOR(S)) ("the Donor(s)") AND MCMASTER UNIVERSITY ("the University") I. THE GIFT (NAME OF DONOR(S)) have generously made a Gift of (X) to McMaster University. This generous Gift will be provided in the form (METHOD OF PAYMENT) according to the following schedule: II

  13. Monitoring human exposure to pesticides using immunoassay

    E-print Network

    Hammock, Bruce D.

    at the estimation of internal dose based on the fate of the compound in human body. Biological monitoring approachesMonitoring human exposure to pesticides using immunoassay Marja E. Koivunen1,2 , Shirley J. Gee1 of automated immunoanalyzers, immunosensors or microchips with flow-through systems. Biological monitoring

  14. Immunoassay on Free-standing Electrospun Membranes

    NASA Astrophysics Data System (ADS)

    Steckl, Andrew; Wu, Dapeng; Han, Daewoo

    2010-03-01

    For the purpose of immunoassay, electrospun membranes can be thought as the thread-like self-assembling of nano/microbeads. Non-woven membranes of electrospun poly(caprolactone) (PCL) fibers display excellent tenacity, flexibility and suitable surface energy. These PCL membranes exhibit easy handling in air, fast spreading and wetting in aqueous solution, and rapid adsorption of protein molecules by hydrophobic interaction. After a fold-and-press process, the membrane porosity was reduced from 75% to less than 10%, while the thickness increased from 5 to 300 ?m. The resulting fluorescence signal from adsorbed protein increased more than 120 times. With anti-HSA and HSA-FITC as an immunoassay model, a linear detection range from 500 ng/mL down to 1 ng/mL is obtained, with a detection of limit (LOD) of 0.08 ng/mL. By comparison, conventional nitrocellulose and thicker PCL fiber electrospun membrane displayed a much higher LOD of 100 ng/mL. Immunoassay on free-standing electrospun membrane successfully combines the low-cost and simplicity of conventional membrane immunoassay, with the fast reaction speed and high sensitivity characteristic of magnetic nano/microbeads bioassays.

  15. IMMUNOASSAY FOR P-NITROPHENOL IN URINE

    EPA Science Inventory

    Urinary excretion of nitrophenol metabolites is an important index of human exposure to organophosphate pesticides. In particular, p-nitrophenol, a major urinary metabolite of parathion, can be used as a biomarker of human exposure. Immunoassay methods have been recently describe...

  16. SITE EVALUATION OF FIELD PORTABLE PENTACHLOROPHENOL IMMUNOASSAYS

    EPA Science Inventory

    Four pentachlorophenol (PCP) enzyme immunoassays for environmental analysis have been evaluated through the U.S. EPA Superfund Innovative Technology Evaluation (SITE) program. Three assays were formatted for on-site field use and one assay could be used in a field laboratory sett...

  17. Immunoassay for Monitoring Environmental and Human Exposure

    E-print Network

    Hammock, Bruce D.

    Immunoassay for Monitoring Environmental and Human Exposure to the Polybrominated Diphenyl Ether) to monitor environmental and human exposure to polybrominated diphenyl ether BDE-47 that is used as a flame workers in electronic-recycling facilities that experience high PBDE exposure (11), a major route

  18. Homogeneous model immunoassay of thyroxine by phase-modulation fluorescense spectroscopy

    NASA Astrophysics Data System (ADS)

    Ozinskas, Alvydas J.; Malak, Henryk M.; Britz, Judy; Thompson, Richard B.; Koen, Peter A.; Lakowicz, Joseph R.

    1993-05-01

    We describe a homogeneous competitive model immunoassay for determination of thyroxine by multi-frequency phase-modulation fluorescence. Using a non-radiative energy transfer transduction mechanism, B-phycoerythrin conjugated to thyroxine is the energy donor and a carboxymethylindocyanine dye conjugated to anti-thyroxine antibody is the energy acceptor. Energy transfer from B-phycoerythrin to the acceptor results in a decreased lifetime and/or phase angle. The fluorescence lifetime change reflects the extent of energy transfer. In the competitive immunoassay format, the donor-thyroxine conjugate and an analytical sample of thyroxine compete for acceptor-antibody binding sites, resulting in a phase angle change which is dependent on the amount of thyroxine in the sample. Dose response curves of phase angle versus thyroxine concentration demonstrate a broader dynamic range than comparable steady state intensity curves. Since phase-modulation lifetime measurements are largely independent of total signal intensity, sources of optical interference are minimized. The potential for whole blood measurements exists since the energy transfer lifetime method can be extended to longer wavelengths.

  19. O-Glycosyl Donors

    NASA Astrophysics Data System (ADS)

    L髉ez, J. Crist骲al

    O-Glycosyl donors, despite being one of the last successful donors to appear, have developed themselves into a burgeoning class of glycosyl donors. They can be classified in two main types: O-alkyl and O-aryl (or hetaryl) glycosyl donors. They share, however, many characteristics, they can be (1) synthesized from aldoses, either by modified Fisher glycosidation (O-alkyl) or by nucleophilic aromatic substitution (O-aryl or O-hetaryl), (2) stable to diverse chemical manipulations, (3) directly used for saccharide coupling, and (4) chemoselectively activated. Among these, n-pentenyl glycosides stand apart. They were the first O-alkyl glycosyl donors to be described and have paved the way to many conceptual developments in oligosaccharide synthesis. The development of the chemoselectivity-based "armed-disarmed" approach for saccharide coupling, including its stereoelectronic or torsional variants, now extended to other kinds of glycosyl donors, was first recognized in n-pentenyl glycosides. The chemical manipulation of the anomeric substituent in the glycosyl donor to induce reactivity differences between related species (sidetracking) was also introduced in n-pentenyl glycosides. An evolution of this concept, the "latent-active" strategy for glycosyl couplings, first described in thioglycosyl donors (vide infra), has been elegantly applied to O-glycosyl donors. Thus, allyl and vinyl glycosides, 2-(benzyloxycarbonyl)benzyl (BCB) glycosides and 2'-carboxybenzyl (CB) glycosides are useful "latent-active" glycosyl pairs. Finally, unprotected 3-methoxy-2-pyridyl (MOP) glycosides have been used in glycosylation processes with moderate success.

  20. GIFT AGREEMENT BETWEEN (NAME OF DONOR(S))

    E-print Network

    Thompson, Michael

    1 GIFT AGREEMENT BETWEEN (NAME OF DONOR(S)) ("the Donor(s)") AND MCMASTER UNIVERSITY ("the University") I. THE GIFT (DONORS NAME(S) have generously made a gift of (GIFT/PLEDGE AMOUNT) to McMaster University (hereinafter the "Gift"). This Gift/Pledge will be provided in the form of (METHOD OF PAYMENT

  1. Theoretical limitations of quantification for noncompetitive sandwich immunoassays.

    PubMed

    Woolley, Christine F; Hayes, Mark A; Mahanti, Prasun; Douglass Gilman, S; Taylor, Tom

    2015-11-01

    Immunoassays exploit the highly selective interaction between antibodies and antigens to provide a vital method for biomolecule detection at low concentrations. Developers and practitioners of immunoassays have long known that non-specific binding often restricts immunoassay limits of quantification (LOQs). Aside from non-specific binding, most efforts by analytical chemists to reduce the LOQ for these techniques have focused on improving the signal amplification methods and minimizing the limitations of the detection system. However, with detection technology now capable of sensing single-fluorescence molecules, this approach is unlikely to lead to dramatic improvements in the future. Here, fundamental interactions based on the law of mass action are analytically connected to signal generation, replacing the four- and five-parameter fittings commercially used to approximate sigmoidal immunoassay curves and allowing quantitative consideration of non-specific binding and statistical limitations in order to understand the ultimate detection capabilities of immunoassays. The restrictions imposed on limits of quantification by instrumental noise, non-specific binding, and counting statistics are discussed based on equilibrium relations for a sandwich immunoassay. Understanding the maximal capabilities of immunoassays for each of these regimes can greatly assist in the development and evaluation of immunoassay platforms. While many studies suggest that single molecule detection is possible through immunoassay techniques, here, it is demonstrated that the fundamental limit of quantification (precision of 10% or better) for an immunoassay is approximately 131 molecules and this limit is based on fundamental and unavoidable statistical limitations. PMID:26342315

  2. Rich Donors, Poor Countries

    ERIC Educational Resources Information Center

    Thomas, M. A.

    2012-01-01

    The shifting ideological winds of foreign aid donors have driven their policy towards governments in poor countries. Donors supported state-led development policies in poor countries from the 1940s to the 1970s; market and private-sector driven reforms during the 1980s and 1990s; and returned their attention to the state with an emphasis on

  3. Donor Telomere Length SAA

    Cancer.gov

    A new NCI study has found that, among patients with severe aplastic anemia who received a hematopoietic cell transplant from an unrelated donor, those whose donor white blood cells had longer telomeres had higher survival rates five-years after transplant

  4. Enzyme immunoassay system for panel testing.

    PubMed

    Donohue, J; Bailey, M; Gray, R; Holen, J; Huang, T M; Keevan, J; Mattimiro, C; Putterman, C; Stalder, A; Defreese, J

    1989-09-01

    An immunoassay system based on enzyme immunoassay technology has been developed for quantitative panel testing. The system includes test card disposables, reagents, and an instrument. Patients' samples are processed semiautomatically in the instrument with minimum user intervention. The test card has multiple test areas at individual locations on a membrane solid phase so that simultaneous determinations from a single specimen are possible. Each panel also includes positive and negative reagent procedural controls. Factory-determined calibration curves for each analyte are provided in barcode form with each test kit. The reagents include a specimen dilution buffer, enzyme conjugate, and precipitogenic substrate. Up to 10 test cards at a time can be processed in random-access and continuous-access modes, with automated agitation of sample and reagents over the solid phase, temperature-controlled incubation, and membrane washing and reading, data reduction, and printout of results. The optical reader measures diffuse reflectance and features source intensity and wavelength compensation. PMID:2673584

  5. Laboratory diagnosis of syphilis with automated immunoassays.

    PubMed

    Marangoni, Antonella; Moroni, Alessandra; Accardo, Silvia; Cevenini, Roberto

    2009-01-01

    The serological detection of specific antibodies to Treponema pallidum is of particular importance in the diagnosis of syphilis. The purpose of this study was to evaluate diagnostic performances of automated immunoassays in comparison with T. pallidum hemagglutination test (TPHA) and Western Blot (WB). The retrospective study was performed with different panels of sera: 244 clinical and serological characterized syphilitic sera and 203 potentially interfering samples. All the sera were tested by Enzygnost Syphilis, ARCHITECT Syphilis TP, TPHA, and homemade WB. The diagnostic performances of the two assays were very similar: both Enzygnost Syphilis and ARCHITECT Syphilis TP performed with a sensitivity of 99.2%, whereas the specificity was 98.5 and 98.4%, respectively. Considering the suitability for automation, both immunoassays may represent a good choice as a screening test. However, the use of a confirmatory test, such as TPHA or WB, remains a must in order to avoid false-positive results. PMID:19140205

  6. Antinuclear antibody detection by automated multiplex immunoassay in untreated patients at the time of diagnosis.

    PubMed

    Op De Be閏k, Katrijn; Vermeersch, Pieter; Verschueren, Patrick; Westhovens, Ren; Mari雗, Godelieve; Blockmans, Daniel; Bossuyt, Xavier

    2012-12-01

    Fully automated multiplex immunoassays are increasingly used as first line screening for antinuclear antibodies. The diagnostic performance of such multiplex assays in untreated patients at the time of diagnosis has not been reported. Antinuclear antibodies were measured by indirect immunofluorescence (IIF) (dilution 1:160) and by BioPlex 2200 ANA screen (antibodies to dsDNA, chromatin, ribosomal protein, SSA-52, SSA-60, SSB, Sm, SmRNP, RNP-A, RNP-68, Scl-70, Jo-1, and centromere B) in 236 patients with a systemic rheumatic disease at the time of diagnosis, 149 blood donors, 139 patients with chronic fatigue syndrome (CFS), and 134 diseased controls. BioPlex ANA screen and IIF were positive in, respectively, 79% and 90% of patients with systemic lupus erythematosus (SLE), 60% and 60% with cutaneous lupus, 72% and 93% with systemic sclerosis (SSc), 100% and 100% with mixed connective tissue disease (MCTD), 89% and 56% with primary Sj鰃ren's (SS) syndrome, 36% and 36% with polymyositis/dermatomyositis, 5.4% and 6% of blood donors, 7.2% and 3.6% of patients with CFS, and 11% and 18% of diseased controls. BioPlex test result interval specific likelihood ratios increased with increasing antibody concentration. The simultaneous presence of at least three antibodies by BioPlex was found in 35% of patients with SLE, 4% with SSc, 100% with MCTD, 64% with SS, 7% with inflammatory myopathy, 0.7% of CFS and diseased controls, and none of the blood donors. In conclusion, test result specific likelihood ratios and the presence of multiple autoantibodies help with the interpretation of data generated by multiplex immunoassays. PMID:22387973

  7. Homogeneous Immunoassays: Historical Perspective and Future Promise

    NASA Astrophysics Data System (ADS)

    Ullman, Edwin F.

    1999-06-01

    The founding and growth of Syva Company is examined in the context of its leadership role in the development of homogeneous immunoassays. The simple mix and read protocols of these methods offer advantages in routine analytical and clinical applications. Early homogeneous methods were based on insensitive detection of immunoprecipitation during antigen/antibody binding. The advent of reporter groups in biology provided a means of quantitating immunochemical binding by labeling antibody or antigen and physically separating label incorporated into immune complexes from free label. Although high sensitivity was achieved, quantitative separations were experimentally demanding. Only when it became apparent that reporter groups could provide information, not only about the location of a molecule but also about its microscopic environment, was it possible to design practical non-separation methods. The evolution of early homogenous immunoassays was driven largely by the development of improved detection strategies. The first commercial spin immunoassays, developed by Syva for drug abuse testing during the Vietnam war, were followed by increasingly powerful methods such as immunochemical modulation of enzyme activity, fluorescence, and photo-induced chemiluminescence. Homogeneous methods that quantify analytes at femtomolar concentrations within a few minutes now offer important new opportunities in clinical diagnostics, nucleic acid detection and drug discovery.

  8. Sequential injection immunoassay for environmental measurements.

    PubMed

    Soh, Nobuaki; Tanaka, Mayumi; Hirakawa, Koji; Zhang, RuiQi; Nakajima, Hizuru; Nakano, Koji; Imato, Toshihiko

    2011-01-01

    Sequential injection immunoassay systems for environmental measurements based on the selective immunoreaction between antigen and antibody were described. A sequential injection analysis (SIA) technique is suitable to be applied for the procedure of enzyme-linked immunosorbent assay (ELISA), because the washing and the addition of reagent solutions can be automated by using a computer-controlled syringe pump and switching valve. We selected vitellogenin (Vg), which is a biomarker for evaluating environmental risk caused by endocrine-disrupting chemicals in the hydrosphere, and linear alkylbenzene sulfonates (LAS) and alkylphenol polyethoxylates (APEO), which are versatile surfactants, as target analytes in the flow immunoassay systems. For Vg monitoring, SIA systems based on spectrophotometric, chemiluminescence, and electrochemical determinations were constructed. On the other hand, chemiluminescence determination was applied to the detection of LAS and APEO. For APEO, an SIA system combined with surface plasmon resonance (SPR) sensor was also developed. These new sequential injection immunoassay systems are expected to be useful systems for environmental analysis. PMID:22076332

  9. Immunoassay as a screening tool for industrial toxicants

    SciTech Connect

    Pierce, T.

    1986-08-01

    Immunoassay techniques may represent useful screening tools to assist analysts interested in the presence and amounts of organic toxicants in biological fluids. The widespread application of immunoassay methods in medicinal and forensic (drugs of abuse) chemistry has resulted in such screening methodologies. Four methodologies of potential benefit are considered: the free radical assay technique, the enzyme-mediated immunoassay technique, radioimmunoassay, and hemagglutination. Each of these immunoassays is based on the competitive displacement of the labeled drug (or toxicant) from the antibody complex by the unlabeled drug-toxicant in the sample.

  10. Living Donor Liver Transplantation

    MedlinePLUS

    ... around the scar. The bulges can usually be fixed with surgery. During your medical exam, ask the ... to find out if the donor's blood type matches the recipient抯 blood type. Next, the transplant team ...

  11. Nanomaterial Labels in Electrochemical Immunosensors and Immunoassays

    SciTech Connect

    Liu, Guodong; Lin, Yuehe

    2007-12-15

    This article reviews recent advances in nanomaterial labels in electrochemical immunosensors and immunoassays. Various nanomaterial labels are discussed, including colloidal gold/silver, semiconductor nanoparticles, and markers loaded nanocarriers (carbon nanotubes, apoferritin, silica nanoparticles, and liposome beads). The enormous signal enhancement associated with the use of nanomaterial labels and with the formation of nanomaterial朼ntibody-antigen assemblies provides the basis for ultrasensitive electrochemical detection of disease-related protein biomarkers, biothreat agents, or infectious agents. In general, all endeavors cited here are geared to achieve one or more of the following goals: signal amplification by several orders of magnitude, lower detection limits, and detecting multiple targets.

  12. Gliadin Detection in Food by Immunoassay

    NASA Astrophysics Data System (ADS)

    Grant, Gordon; Sporns, Peter; Hsieh, Y.-H. Peggy

    Immunoassays are very sensitive and efficient tests that are commonly used to identify a specific protein. Examples of applications in the food industry include identification of proteins expressed in genetically modified foods, allergens, or proteins associated with a disease, including celiac disease. This genetic disease is associated with Europeans and affects about one in every 200 people in North America. These individuals react immunologically to wheat proteins, and consequently their own immune systems attack and damage their intestines. This disease can be managed if wheat proteins, specifically "gliadins," are avoided in foods.

  13. Fluorimetric Immunoassay for Multianalysis of Aflatoxins

    PubMed Central

    2013-01-01

    A sensitive fluorimetric ELISA was developed for the analysis of aflatoxins. The assay was performed in a 384 microwell plate, wherein high specificity monoclonal antibody against AFM1 (mAb-AFM1) was used as capture antibody and FITC conjugated secondary antibody was used for detection and quantification of the analyte. The linear range of the immunoassay was found to be 6.2550?pg/mL. AFM1 as low as 1?pg/mL was detected by this method with assay volume 40??L. The multi-analysis of different aflatoxins was also investigated in the microwell plate, based on the cross-reactivity (CR) approach. Real milk samples were tested along with certified reference material by standard addition method and recovery analysis was done. The mAb-AFM1 showed 23.2% CR with AFB1, 50% CR with respect to AFM2, and least CR towards AFG1 (<1%). Furthermore, mixture analysis of AFM2 and AFB1 was carried out at specific concentrations of AFM1. The advantages of this developed immunoassay are high sensitivity, high throughput, multianalyte detection, versatility, and ease of handling. PMID:24000318

  14. Dialing for Donors

    ERIC Educational Resources Information Center

    Schaffhauser, Dian

    2012-01-01

    When times get tough, grown children often turn to their parents for help--for some extra cash, even somewhere to stay. For colleges and universities, that role is filled by alumni donors. In 2011, with education budgets slashed across the country, giving accounted for 6.5 percent of college expenditures, according to the Council for Aid to

  15. Donation Form DONOR INFORMATION

    E-print Network

    Montr閍l, Universit de

    Donation Form DONOR INFORMATION First and Last Names: Organization Name: Address: Contact Name: Email: ONE DONATION, 5 WAYS TO MAKE A DIFFERENCE 1. Support students in their educational needs for student scholarships. This will be ensured through the placement of 80% of all donations

  16. Health risk of leukapheresis donors.

    PubMed

    Sandler, S G; Nusbacher, J

    1982-01-01

    The increasing use of normal donors for leukapheresis to provide granulocyte concentrates requires careful monitoring of recruitment and operational procedures to minimize donor health risks. Of 18,288 leuka- and leukaplateletphereses performed in American Red Cross regional blood services during a 12-month period, only 0.27% were discontinued because of acute donor reactions. None of these reactions had serious or long-term consequences for the donors. The potential toxicities of corticosteroids and hydroxyethyl starch, and the theoretical hazards of lymphocyte depletion, raise unanswerable questions concerning long-term safety, although no serious adverse reactions related to these factors have been observed. Critical factors for leukapheresis donor safety are: selective donor recruitment, adherence to operational protocols, and continuous and informed interest of physicians responsible for pheresis programs. Additional data are needed to assess the effects of repeated leukapheresis on donors and to establish rational guidelines for donor recruitment. PMID:6180961

  17. Immunoassay procedures for fiber optic sensors

    NASA Astrophysics Data System (ADS)

    Ligler, Frances S.

    1988-04-01

    There is an increasing need for the development of an ultrasensitive immunoassay for use with fiber optic sensors. These detection systems can be used for such applications as disease diagnosis, detection of chemical and biological warfare agents or drugs of abuse, pollution control, therapeutic monitoring, and explosive detection. This specific program is designed to produce generic chemistries for use with existing fiber optic-based sensors to detect pathogens of particular threat to Army personnel as determined by USAMRIID. The detection system under development involves the attachment of antibodies to an optical fiber at high density. In addition, the immobilization must be achieved in a way which retains the antibody's ability to bind antigen. The functionality of the antibody will be tested through the binding of a labelled antigen. In the future, this assay could incorporate the antibodies developed by the Army for pathogens of particularly military concern.

  18. Nanoparticles for Enhanced Sensitivity in Electrochemical Immunoassays

    SciTech Connect

    Lin, Yuehe; Wang, Jun; Wang, Hua; Wu, Hong; Tang, Zhiwen

    2008-10-12

    In this manuscript, we report on electrochemical biosensors based on various nanoparticles (NPs) as labels for sensitive detection of protein biomarkers. We used silica nanoparticle as a carrier to loading a large amount of electroactive species such as poly(guanine) for sensitive immunoassay of tumor necrosis factor-alpha (TNF-a). We took the advantages of the unique hollow structure and reconstruction properties of apoferritin to prepare Cd3(PO4)2 nanoparticles as labels for sensitive assay of TNF-a. A novel immunochromatographic/electro-chemical biosensor based on quantum dots as labels has also been developed for rapid and sensitive detection of prostate-specific antigen (PSA) in human serum. These biosensors are quite sensitive with the detection limit at pM level and these approaches based on nanoparticle labels offer a new avenue for sensitive detection of protein biomarkers.

  19. DETECTION OF ROTAVIRUS WITH A NEW POLYCLONAL ANTIBODY ENZYME IMMUNOASSAY (ROTAZYME 2) AND A COMMERCIAL LATEX AGGLUTINATION TEXT (ROTALEX): COMPARISON WITH A MONOCLONAL ANTIBODY ENZYME IMMUNOASSAY

    EPA Science Inventory

    A total of 176 human fecal specimens were examined for the presence of rotavirus using four different assays: a monoclonal antibody enzyme immunoassay; the original polyclonal antibody enzyme immunoassay marketed by Abbott Laboratories, Chicago, IL (Rotazyme I); a modification of...

  20. Detection of narcotics with an immunoassay film badge

    SciTech Connect

    Lukens, H.R.

    1993-12-31

    Efficient personnel performance, a major requirement for a safe nuclear industry, is jeopardized where personnel use narcotics. However, detection of narcotics at nuclear plants is a challenge. The unique specificity and sensitivity of an immunoassay has been implemented in the form of a small, dry immunoassay film badge (IFB) for the detection of vapors emitted by narcotics. The device is suitable as an area monitor, and its characteristics are suitable for use as a breath monitor for the detection of drug use.

  1. Capillary Electrophoresis-Based Immunoassays: Principles & Quantitative Applications

    PubMed Central

    Moser, Annette C.; Hage, David S.

    2013-01-01

    The use of capillary electrophoresis as a tool to conduct immunoassays has been an area of increasing interest over the last decade. This approach combines the efficiency, small sample requirements, and relatively high speed of CE with the selectivity of antibodies as binding agents. This review examines the various assay formats and detection modes that have been reported for these assays, along with some representative applications. Most CE immunoassays in the past have employed homogeneous methods in which the sample and reagents are allowed to react in solution. These homogeneous methods have been conducted as both competitive binding immunoassays and as non-competitive binding immunoassays. Fluorescent labels are most commonly used for detection in these assays, but enzyme labels have also been utilized for such work. Some additional work has been performed in CE immunoassays with heterogeneous methods in which either antibodies or an analog of the analyte is immobilized to a solid support. These heterogeneous methods can be used for the selective isolation of analytes prior to their separation by CE or to remove a given species from a sample/reagent mixture prior to analysis by CE. These CE immunoassays can be used with a variety of detection modes, such as fluorescence, UV/visible absorbance, chemiluminescence, electrochemical measurements, mass spectrometry, and surface plasmon resonance. PMID:18646279

  2. Clinical Applications of Capillary Electrophoresis-Based Immunoassays

    PubMed Central

    Moser, Annette C.; Willicott, Corey W.; Hage, David S.

    2014-01-01

    Immunoassays have long been an important set of tools in clinical laboratories for the detection, diagnosis and treatment of disease. Over the last two decades there has been growing interest in utilizing capillary electrophoresis (CE) as a means for conducting immunoassays with clinical samples. The resulting method is known as a CE immunoassay. This approach makes use of the selective and strong binding of antibodies for their targets, as is employed in a traditional immunoassay, and combines this with the speed, efficiency, and small sample requirements of CE. This review discusses the variety of ways in which CE immunoassays have been employed with clinical samples. An overview of the formats and detection modes that have been employed in these applications is first presented. A more detailed discussion is then given on the type of clinical targets and samples that have been measured or studied by using CE immunoassays. Particular attention is given to the use of this method in the fields of endocrinology, pharmaceutical measurements, protein and peptide analysis, immunology, infectious disease detection, and oncology. Representative applications in each of these areas are described, with these examples involving work with both traditional and microanalytical CE systems. PMID:24132682

  3. Detection of poliovirus antigen by enzyme immunoassay.

    PubMed Central

    Ukkonen, P; Huovilainen, A; Hovi, T

    1986-01-01

    A solid-phase enzyme immunoassay (EIA) was developed for the detection of poliovirus antigen. Rabbit and guinea pig antisera for the assay were raised against purified poliovirus type 3/Fin (strain 3/Fin/K) isolated from a fecal specimen from a meningitis patient during an outbreak of poliomyelitis in Finland in 1984. The EIA was highly specific for poliovirus type 3, and it was about 30 times more sensitive for strain 3/Fin/K than for strain 3/Saukett used in the inactivated poliovirus vaccine. The sensitivity of the EIA was 2 to 5 ng of purified strain 3/Fin/K per ml, whereas disrupted viruses and soluble viral proteins were almost undetectable by the assay. Only 5 of 51 (10%) stool specimens containing poliovirus type 3/Fin detected by virus isolation were positive by the EIA. Quantitation by the EIA, using purified poliovirus 3/Fin/K as a standard, revealed that concentrations of poliovirus type 3 in undiluted fecal specimens of patients with natural poliovirus infection were only 50 ng/ml or less. In conclusion, owing to the small amount of poliovirus in feces, the EIA is not suitable for the diagnosis of poliovirus infections directly from clinical specimens, but it can be used to detect, type, and quantitate poliovirus antigen in infected cells. PMID:3023440

  4. Performance characteristics of eight estradiol immunoassays.

    PubMed

    Yang, David T; Owen, William E; Ramsay, Carol S; Xie, Hui; Roberts, William L

    2004-09-01

    Measurement of estradiol is useful in assisted reproduction, evaluation of infertility, menopause, and male feminization. The analytic performance of 8 estradiol immunoassays was evaluated. The imprecision and accuracy of the Access, ADVIA Centaur, ARCHITECT i2000, AutoDELFIA, Elecsys 2010, IMMULITE 2000, and Vitros ECi estradiol assays (see text for proprietary information) were evaluated by using an isotope dilution-gas chromatography-mass spectrometry (ID-GC-MS) reference method. The coefficient of variation (CV) ranged from 6.9% on the Elecsys 2010 to 42.6% on the ADVIA Centaur at an estradiol concentration of 18 pg/mL (66 pmol/L), with the ARCHITECT i2000 assay in development and the Vitros ECi having a CV below 10% at this estradiol concentration. Agreement between the automated assays and ID-GC-MS was variable, with slopes ranging from 0.87 to 1.20. The Access, ARCHITECT i2000 in development, and the IMMULITE 2000 were the most accurate, with slopes of 0.99, 0.98, and 1.03, respectively. These findings indicate that the ARCHITECT i2000 estradiol assay in development had the best precision and accuracy of the assays evaluated for measurement of serum estradiol concentrations. PMID:15362362

  5. Seroprevalence of human T lymphotropic virus antibodies among healthy blood donors at a tertiary centre in Lagos, Nigeria

    PubMed Central

    Durojaiye, Idris; Akinbami, Akinsegun; Dosunmu, Adedoyin; Ajibola, Sarah; Adediran, Adewumi; Uche, Ebele; Oshinaike, Olajumoke; Odesanya, Majeed; Dada, Akinola; Okunoye, Olaitan

    2014-01-01

    Introduction Transmission of human T-lymphotropic viruses (HTLV) occurs from mother to child, by sexual contact and blood transfusion. Presently, in most centres in Nigeria, there is no routine pre-transfusion screening for HTLV. The study aims to determine the prevalence of HTLV-1 and HTLV-2 among healthy blood donors at a tertiary centre in Lagos. Methods A cross-sectional study was carried out at the blood donor clinic of the Lagos State University Teaching Hospital (LASUTH), Ikeja. About 5mls of venous blood was collected from each subject into a sterile plain bottle after obtaining subject's consent. The serum separated and stored at -200C. Sera were assayed for HTLV by an enzyme-linked immunoassay (ELISA) for the determination of antibodies to HTLV 1 and HTLV -2. Western blot confirmatory testing was done on reactive samples. All donors were also screened for HIV, HBsAg and HCV by rapid kits. Results The seroprevalence of HTLV -1 by ELISA was 1.0% and 0.5% by Western Blot among blood donors. A total of 210 healthy blood donors were enrolled. Only 2 (1.0%) blood donors were repeatedly reactive with ELISA test. On confirmatory testing with Western Blot, 1 (0.5%) blood donor was positive for HTLV. All the healthy blood donors were negative for HIV, HbsAg and HCV. None of the 210 blood donors had been previously transfused; as such no association could be established between transfusion history and HTLV positivity among the blood donors. Conclusion The seroprevalence of HTLV in this environment is low among healthy blood donors. PMID:25328597

  6. Capillary Isoelectric Focusing Immunoassay for Fat Cell Differentiation Proteomics

    PubMed Central

    Johlfs, Mary G.; Gorjala, Priyatham; Urasaki, Yasuyo; Le, Thuc T.; Fiscus, Ronald R.

    2015-01-01

    Profiling cellular proteome is critical to understanding signal integration during cell fate determination. In this study, the capability of capillary isoelectric focusing (cIEF) immunoassays to detect post-translational modifications (PTM) of protein isoforms is demonstrated. cIEF immunoassays exhibit protein detection sensitivity at up to 5 orders of magnitude higher than traditional methods. This detection ultra-sensitivity permits proteomic profiling of several nanograms of tissue samples. cIEF immunoassays are employed to simultaneously profile three protein kinases during fat cell differentiation: cGMP-dependent protein kinase type I (PKG-I) of the nitric oxide (NO) signaling pathway, protein kinase B (Akt) of the insulin signaling pathway, and extracellular signal-regulated kinase (ERK) of the mitogen-activated protein kinase (MAPK) signaling pathway. Interestingly, a switch in the expression level of PKG- isoforms is observed during fat cell differentiation. While both PKG-I? and PKG-I? isoforms are present in preadipocytes, only PKG-I? isoform is expressed in adipocytes. On the other hand, the phosphorylation level increases for Akt while decreases for ERK1 and ERK2 following the maturation of preadipocytes into adipocytes. Taken together, cIEF immunoassay provides a highly sensitive means to study fat cell differentiation proteomics. cIEF immunoassay should be a powerful proteomics tool to study complex protein signal integration in biological systems. PMID:26132171

  7. Trends in Transfusion Transmitted Infections Among Replacement Blood Donors in Karachi, Pakistan

    PubMed Central

    Irfan, Syed Mohammad; Uddin, Jamal; Zaheer, Hasan Abbas; Sultan, Sadia; Baig, Amjad

    2013-01-01

    Objective: To determine the prevalence of Hepatitis-B, Hepatitis-C and Human Immunodeficiency infections in replacement blood donors. Materials and Methods: From January 2004 to December 2011, 108,598 apparently healthy donors donated blood at our Blood Bank. Screening was done by Microparticle Enzyme Immuno Assay (MEIA) method on Axsym System (Abbott Diagnostic, USA) and in year 2011 by Chemiluminescent Immunoassay (CIA) method on Architect i2000 (Abbott Diagnostic, USA). From 2010 onward, HIV reactive donors were advised for confirmatory tests and reported back with the results. Results: Of the 108,598 total donors, 108,393 (99.8%) were replacement donors with a mean age of 28.92 (17-55) years. Of this, only 164 (0.15%) were females. Among the replacement donors, 4,906 (4.5%) were found to be reactive for Hepatitis-B, C and Human Immunodeficiency Virus. All the reactive patients, except one, were males. HbsAg was positive in 2,068 (1.90%) and anti-HCV in 2832 (2.61%) donors, while 111 (0.10%) were positive for Human Immunodeficiency Virus. Co-infectivity was observed in 103 (0.09%) cases. The prevalence appeared to be higher in younger age group (17-30 yrs). Only 16.6% cases should be patients returned with results of the confirmatory tests for HIV and were found positive. Conclusion: Hepatitis-B and C sero-prevalence in our series of replacement donors appears high compared to most studies from neighboring countries and relatively low in comparison to earlier studies from Pakistan. Prevalence of HIV, however, appears low and turn out of HIV positive cases for confirmatory tests is low. Conflict of interest:None declared. PMID:24385780

  8. SUPERFUND INNOVATIVE TECHNOLOGY EVALUATION (SITE) REPORT FOR THE WESTINGHOUSE BIOANALYTICAL SYSTEMS PENTACHLOROPHENOL (PCP) IMMUNOASSAYS

    EPA Science Inventory

    The results of the demonstration of two Westinghouse Bio-Analytic Systems (WBAS) immunoassay technologies are described in this report. The immunoassays measure parts per billion concentrations of pentachlorophenol in environmental water samples. The study was conducted under the...

  9. SUPERFUND INNOVATIVE TECHNOLOGY EVALUATION PROGRAM DEMONSTRATION PLAN FOR WESTINGHOUSE BIO-ANALYTIC SYSTEMS PENTACHLOROPHENOL IMMUNOASSAYS

    EPA Science Inventory

    This plan provides a detailed design and description of the demonstration and evaluation program for the Westinghouse Bio-Analytic Systems immunoassay technologies specific for the analysis of pentachlorophenol. he immunoassays measure parts per billion concentrations of pentachl...

  10. The Effects of Sample Matrices on Immunoassays to Detect Microcystin-LR in Water

    EPA Science Inventory

    Abstract: Immunoassays are widely used biochemical techniques to detect microcystins in environmental samples. The use of immunoassays for the detection of microcystins is vulnerable to matrix components and other interferents. This study is an evaluation of the effects of interf...

  11. PRNP variants in goats reduce sensitivity of detection of PrPSc by immunoassay

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Immunoassays are extensively utilized in disease diagnostics with monoclonal antibodies serving as critical tools within the assay. Detection of scrapie in sheep and goats relies heavily on immunoassays including immunohistochemistry, western blotting, and ELISA. In the United States, regulatory tes...

  12. Organ Donor FAQ's: Who Can Be a Donor

    MedlinePLUS

    ... citizens have been organ donors. Can non-resident aliens donate and receive organs? Non-resident aliens can both donate and receive organs in the ... the 12,375 organ donors were non-resident aliens. In this same year, 259 (1%) of the ...

  13. A Wash-Free Homogeneous Colorimetric Immunoassay Method

    PubMed Central

    Liu, Huiqiao; Rong, Pengfei; Jia, Hongwei; Yang, Jie; Dong, Bo; Dong, Qiong; Yang, Cejun; Hu, Pengzhi; Wang, Wei; Liu, Haitao; Liu, Dingbin

    2016-01-01

    Rapid and convenient biosensing platforms could be beneficial to timely diagnosis and treatment of diseases in virtually any care settings. Sandwich immunoassays, the most commonly used methods for protein detection, often rely on expensive tags such as enzyme and tedious wash and incubation procedures operated by skilled labor. In this report, we revolutionized traditional sandwich immunoassays by providing a wash-free homogeneous colorimetric immunoassay method without requirement of any separation steps. The proposed strategy was realized by controlling the growth of gold nanoparticles (AuNPs) to mediate the interparticle spacing in the protein-AuNP oligomers. We have demonstrated the successful in vitro detection of cancer biomarker in serum samples from patients with high clinical sensitivity and specificity. PMID:26722373

  14. A Wash-Free Homogeneous Colorimetric Immunoassay Method.

    PubMed

    Liu, Huiqiao; Rong, Pengfei; Jia, Hongwei; Yang, Jie; Dong, Bo; Dong, Qiong; Yang, Cejun; Hu, Pengzhi; Wang, Wei; Liu, Haitao; Liu, Dingbin

    2016-01-01

    Rapid and convenient biosensing platforms could be beneficial to timely diagnosis and treatment of diseases in virtually any care settings. Sandwich immunoassays, the most commonly used methods for protein detection, often rely on expensive tags such as enzyme and tedious wash and incubation procedures operated by skilled labor. In this report, we revolutionized traditional sandwich immunoassays by providing a wash-free homogeneous colorimetric immunoassay method without requirement of any separation steps. The proposed strategy was realized by controlling the growth of gold nanoparticles (AuNPs) to mediate the interparticle spacing in the protein-AuNP oligomers. We have demonstrated the successful in vitro detection of cancer biomarker in serum samples from patients with high clinical sensitivity and specificity. PMID:26722373

  15. Donor Preferences and Charitable Giving

    ERIC Educational Resources Information Center

    Williams, Stephanie Roderick

    2007-01-01

    This study aimed to learn more of the differences that may exist between the two most powerful groups of donors today, baby boomers (40-58 years old) and mature donors (59 and older), in an effort to help organizations improve fundraising efforts. Questions about the importance of organizational efficiency, program outcomes, and the desire for

  16. Management of Young Blood Donors

    PubMed Central

    Newman, Bruce H.

    2014-01-01

    Summary The emphasis on high-school blood drives and acceptance of 16-year-old blood donors led to more research on physiologic and psychological ways to decrease vasovagal reaction rates in young blood donors and to increase donor retention. Research on how to accomplish this has been advantageous for the blood collection industry and blood donors. This review discussed the current situation and what can be done psychologically, physiologically, and via process improvements to decrease vasovagal reaction rates and increase donor retention. The donation process can be significantly improved. Future interventions may include more dietary salt, a shorter muscle tension program to make it more feasible, recommendations for post-donation muscle tension / squatting / laying down for lightheadedness, more donor education by the staff at the collection site, more staff attention to donors with fear or higher risk for a vasovagal reaction (e.g. estimated blood volume near 3.5 l, first-time donor), and a more focused donation process to ensure a pleasant and safer procedure. PMID:25254024

  17. Marrow transplantation from unrelated donors.

    PubMed

    Sierra, J; Anasetti, C

    1995-11-01

    The use of an HLA-compatible unrelated donor is an option for patients who require an allogeneic transplant but lack a family member match. Grafts from unrelated volunteer donors have provided long-term disease-free survival for a variable proportion of patients, depending on degree of HLA matching with the donor, patient's disease, disease stage, and age. The number of volunteers in marrow donor registries worldwide has increased to more than 2.5 million. The number of unrelated donor transplants facilitated by the US National Marrow Donor Program alone will exceed 900 this year. Progress in HLA-typing technology results in a more precise definition of donor and recipient matching and new assays have been developed with initial success to measure alloreactive T-cell precursors for selection of donors with less antihost reactivity. Prevention and treatment of graft failure, graft-versus-host disease, opportunistic infections, and Epstein-Barr virus-associated lymphoproliferative disease remain a challenge. PMID:9372034

  18. Protein microchips : use for immunoassay and enzymatic reactions.

    SciTech Connect

    Arenkov, P.; Kukhtin, A.; Gemmell, A.; Voloschuk, S.; Chupeeva, V.; Mirzabekov, A.; Biochip Technology Center; Russian Academy of Sciences

    2000-02-15

    Different proteins such as antibodies, antigens, and enzymes were immobilized within the 100 x 100 x 20-{mu}m gel pads of protein microchips. A modified polyacrylamide gel has been developed to accommodate proteins of a size up to 400,000 daltons. Electrophoresis in the microchip reaction chamber speeded up antigen-antibody interactions within the gel. Protein microchips were used in immunoassays for detection of antigens or antibodies, as well as to carry out enzymatic reactions and to measure their kinetics in the absence or presence of an inhibitor. A protein microchip can be used several times in different immunoassays and enzymatic kinetic measurements.

  19. Variability in Telavancin Cross-Reactivity among Vancomycin Immunoassays

    PubMed Central

    McConeghy, Kevin W.; Liao, Siyun; Clark, Douglas; Worboys, Philip; Barriere, Steven L.

    2014-01-01

    Telavancin is a semisynthetic lipoglycopeptide with a dual mechanism of action against Gram-positive pathogens. Two brief reports have suggested potential cross-reactivity of telavancin with the vancomycin particle-enhanced turbidometric immunoassay (PETIA). The purpose of this study was to evaluate several commercially available vancomycin immunoassays (fluorescence polarization [FPIA], enzyme-multiplied immunoassays [EMIT], PETIA, and chemiluminescent immunoassay [CMIA]) for cross-reactivity with telavancin. Seven sites were selected to analyze serum samples for vancomycin. Each site received a set of samples (n = 18) which combined drug-free serum with telavancin, 7-OH telavancin metabolite, or vancomycin. Immunoassays demonstrating potential cross-reactivity were further evaluated by sending a duplicate sample set to multiple laboratories. Cross-reactivity was defined as the percent theoretical concentration (reported concentration/theoretical concentration 100). No cross-reactivity was seen with FPIA or EMIT. Within the theoretical concentration range of 5 to 120 ?g/ml of telavancin, the Synchron PETIA system reported vancomycin concentrations ranging from 4.7 to 54.2 ?g/ml compared to vancomycin concentrations from 1.1 to 5.6 ?g/ml for the Vista PETIA system. The Architect CMIA system reported vancomycin concentrations in the range of 0.27 to 0.97 ?g/ml, whereas Advia Centaur XP CMIA reported vancomycin concentrations between 1.6 and 31.6 ?g/ml. The Architect CMIA immunoassay had the lowest percent cross-reactivity (0.8 to 5.4%), while the Synchron PETIA immunoassay demonstrated the highest percent cross-reactivity (45.2 to 53.8%). Telavancin samples measured by liquid chromatography-mass spectroscopy were within 93.9 to 122% of theoretical concentrations. Vancomycin concentrations were not measured in any 7-OH telavancin-spiked sample. Vancomycin concentrations measured by liquid chromatography-mass spectroscopy were within 57.2 to 113% of theoretical concentrations. PETIA and CMIA measured vancomycin concentrations in telavancin-spiked samples. Significant variability in percent cross-reactivity was observed for each platform regardless of immunoassay method. PMID:25223996

  20. Robust detection of peak signals for lateral flow immunoassays

    NASA Astrophysics Data System (ADS)

    Kim, Jongwon; Kim, Jong Dae; Nahm, Kie Bong; Choi, Eui Yul; Lee, Geumyoung

    2011-02-01

    Template matching method is presented to identify the peaks from the scanned signals of lateral flow immunoassay strips. The template is composed of two pulses separated by the distance of the control and the target ligand line in the assay, and is convolved with the scanned signal to deliver the maximum at the center of the two peaks. The peak regions were identified with the predefined distances from the center. Glycosylated haemoglobin immunoassay strips and fluorescent strip readers from Boditechmed Inc. were tested to estimate the lot and reader variations of the concentration measurands. The results showed the robustness of the propose method.

  1. Monitoring Pesticides and Personal Care Chemicals in Water by Immunoassay

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Due to the increasing number and quantity of organic pollutants, regulatory authorities require implementation of rapid, reliable, and cost-effective technologies for monitoring of water quality. Immunoassays provide a simple, powerful and inexpensive method for monitoring organic contaminants in bo...

  2. Kinase Activity Studied in Living Cells Using an Immunoassay

    ERIC Educational Resources Information Center

    Bavec, Aljos?a

    2014-01-01

    This laboratory exercise demonstrates the use of an immunoassay for studying kinase enzyme activity in living cells. The advantage over the classical method, in which students have to isolate the enzyme from cell material and measure its activity in vitro, is that enzyme activity is modulated and measured in living cells, providing a more

  3. Enzyme linked immunoassay with stabilized polymer saccharide enzyme conjugates

    DOEpatents

    Callstrom, M.R.; Bednarski, M.D.; Gruber, P.R.

    1997-11-25

    An improvement in enzyme linked immunoassays is disclosed wherein the enzyme is in the form of a water soluble polymer saccharide conjugate which is stable in hostile environments. The conjugate comprises the enzyme which is linked to the polymer at multiple points through saccharide linker groups. 19 figs.

  4. A Compact Immunoassay Platform Based on a Multicapillary Glass Plate

    PubMed Central

    Xue, Shuhua; Zeng, Hulie; Yang, Jianmin; Nakajima, Hizuru; Uchiyama, Katsumi

    2014-01-01

    A highly sensitive, rapid immunoassay performed in the multi-channels of a micro-well array consisting of a multicapillary glass plate (MCP) and a polydimethylsiloxane (PDMS) slide is described. The micro-dimensions and large surface area of the MCP permitted the diffusion distance to be decreased and the reaction efficiency to be increased. To confirm the concept of the method, human immunoglobulin A (h-IgA) was measured using both the proposed immunoassay system and the traditional 96-well plate method. The proposed method resulted in a 1/5-fold decrease of immunoassay time, and a 1/56-fold cut in reagent consumption with a 0.05 ng/mL of limit of detection (LOD) for IgA. The method was also applied to saliva samples obtained from healthy volunteers. The results correlated well to those obtained by the 96-well plate method. The method has the potential for use in disease diagnostic or on-site immunoassays. PMID:24859022

  5. Enzyme linked immunoassay with stabilized polymer saccharide enzyme conjugates

    DOEpatents

    Callstrom, Matthew R. (Columbus, OH); Bednarski, Mark D. (Berkeley, CA); Gruber, Patrick R. (St. Paul, MN)

    1997-01-01

    An improvement in enzyme linked immunoassays is disclosed wherein the enzyme is in the form of a water soluble polymer saccharide conjugate which is stable in hostile environments. The conjugate comprises the enzyme which is linked to the polymer at multiple points through saccharide linker groups.

  6. A fluorescence immunoassay for soluble antigens employing flow cytometric detection.

    PubMed

    Lisi, P J; Huang, C W; Hoffman, R A; Teipel, J W

    1982-04-01

    A "sandwich" fluorescence immunoassay is described which does not require the physical separation of bound from free label. Antibody coated microspheres, sample and fluorescent antibody are reacted together as in a conventional 'sandwich' immunoassay except that separation and washing steps are omitted. After the reaction is completed, the suspension is introduced directly into a flow cytometer equipped with a laser light source and both fluorescent and scattered light detection capabilities. By gating fluorescence light accumulation on scattered light pulses, particles associated fluorescence may be selectively measured. The system was evaluated in a model immunoassay for human immunoglobulin (hIgG), employing anti-hIgG coated microspheres (1--5 micrometer and 40--50 micrometer polyacrylamide beads and 30--40 micrometer dextran beads), fluorescein-labeled rabbit anti-hIgG and a Spectrum III flow cytometer. Sensitivities of 10 ng/ml and intra-assay precisions of 2--10% were achieved in a serum matrix. The approach potentially provides a general nonseparation immunoassay format for quantitatively measuring both small and large molecular weight soluble antigens, as well as cell surface antigens. PMID:7039872

  7. Microneutralization test for rabies virus based on an enzyme immunoassay.

    PubMed

    Mannen, K; Mifune, K; Reid-Sanden, F L; Smith, J S; Yager, P A; Sumner, J W; Fishbein, D B; Tong, T C; Baer, G M

    1987-12-01

    We have developed an enzyme immunoassay for rabies virus by using acetone-fixed infected cell cultures as the antigen. This test was used to demonstrate virus-neutralizing antibodies in human and animal sera and was as sensitive as and easier to perform than the rapid fluorescent-focus inhibition technique. PMID:3323234

  8. DETECTION OF NORWALK VIRUS IN STOOLS BY ENZYME IMMUNOASSAY

    EPA Science Inventory

    The development of a solid-phase microtiter enzyme immunoassay (EIA) for detection of Norwalk virus antigen in stool samples is described. The EIA was compared with a previously developed radioimmunoassay (RIA) for detection of Norwalk virus antigen in stools obtained from 30 vol...

  9. Bone density in apheresis donors and whole blood donors.

    PubMed

    Boot, C L; Luken, J S; van den Burg, P J M; de Kort, W L A M; Koopman, M M W; Vrielink, H; van Schoor, N M; den Heijer, M; Lips, P

    2015-11-01

    Apheresis donation using citrate causes acute decrease in serum calcium and increase in serum parathyroid hormone. Long-term consequences, such as decrease in bone mineral density (BMD), are not known. In this study, we compared the BMD of 20 postmenopausal apheresis donors (mean donation number 115 times in up to 15爕ears) with that of 20 whole blood donors (for 15爕ears or more) aged 55-70. BMD in the lumbar spine was not lower in apheresis donors than in blood donors (mean牨燬D 100牨񁖝8 vs. 092牨񁖝2, P=񁖜9). In the hip, BMD was not different between the groups. PMID:26031345

  10. [Altruism and the donor].

    PubMed

    Langlois, A

    1991-08-01

    On December 20, 1988, the government of France passed a law to protect people who voluntarily participate in biomedical research. This article makes extensive reference to a major study, titled From Biology to Ethics, by Jean Bernard, a well-respected authority in the field of bioethics. The author looks at models proposed by Bernard, as examples for health volunteers, in particular, the blood donor and the self-experimenter. To set the tone of the article, she recalls the concept of altruism, as first proposed by Auguste Comte, then makes a linkage between his philosophy and Bernard's point of view. By trial and error, in their discussions, various ethics committees and the French State Council have agreed upon what constitutes fair compensation under the law. Unlike their Canadian counterparts, medical researchers in France have free access to volunteers who are not in perfect health--e.g., the elderly, people suffering from kidney deficiency, cirrhosis of the liver, etc.--but these "experimental subjects" receive no monetary compensation. Thus, healthy and less-than-healthy volunteers do not receive equal treatment under the law. This inequity, added to the fear of what amounts to a tax on the human body and the difficulty of ensuring just compensation, is giving rise to a great deal of uncertainty. PMID:1878857

  11. Comparison of Common Immunoassay Kits for Effective Application in Workplace Drug Urinalysis.

    PubMed

    Liu, R H

    1994-06-01

    Workplace drug urinalysis protocols include an initial immunoassay followed by a confirmation gas chromatography/mass spectrometry (GC/MS) test of immunoassay-positive samples. (Drug categories that are commonly tested include: amphetamines, barbiturates, benzodiazepines, cannabis, cocaine, lysergic acid diethyl amide, methadone, methaqualone, opiates, phencyclidine, and propoxyphene. Not all drug categories are tested by all workplace drug urinalysis programs.) Only those samples that are tested positive by both the initial and the confirmatory procedures can be reported as positive. Thus, when adopting an immunoassay, one must have knowledge of the assay's cross-reacting characteristics and the assay's apparent analyze concentration that corresponds to a specific analyze concentration determined by the GC/MS procedure. The underlying principles of the commonly used radioimmunoassay, enzyme immunoassay, fluorescence polarization immunoassay, and particle immunoassay are outlined. Cross-reacting characteristics of these immunoassays as reported by the manufacturers and independent laboratories are tabulated. This information shown that commercial immunoassay kits for drug categories that are included in workplace drug urinalysis programs are generally more specific than those kits that are for clinical use only. Furthermore, recently manufactured immunoassay kits targeted for use in workplace drug urinalysis programs are more specific than those manufactured earlier. Reported effects of adulterants, such as salt, cleaning agents, etc., on commonly used immunoassays are summarized. Without more comprehensive and systematic studies, it is difficult to make general statements concerning the superiority of one methodology over the others. It is clear, however, that cannabinoid assesses are most susceptible to the influence of adulterants. Reported immunoassay-GC/MS correlation data are reviewed. Significant correlations exist in all cases. The immunoassay apparent analyze concentration corresponding to a specific GC/MS analyze concentration may be approximately based on the resulting regression equations. Since the corresponding immunoassay apparent analyze concentrations vary with the specificities of the reagent used, the immunoassay reagent manufacturers should carefully study specificity characteristics of each manufacturing batch and provide these correlation data for users' evaluation and adaptation. PMID:26270150

  12. How to Motivate Whole Blood Donors to Become Plasma Donors

    PubMed Central

    2014-01-01

    This study tested the efficacy of interventions to recruit new plasma donors among whole blood donors. A sample of 924 donors was randomized to one of three conditions: control; information only by nurse; and information plus self-positive image message by nurse (SPI). Participants in the control condition only received a leaflet describing the plasma donation procedure. In the two experimental conditions the leaflet was explained face-to-face by a nurse. The dependent variables were the proportion of new plasma donors and the number of donations at six months. Overall, 141 (15.3%) new plasma donors were recruited at six months. There were higher proportions of new plasma donors in the two experimental conditions compared to the control condition (P < .001); the two experimental conditions did not differ. Also, compared to the control condition, those in the experimental conditions (all Ps < .001) gave plasma more often (information only by nurse:??d = .26; SPI: d = .32); the SPI intervention significantly outperformed (P < .05) the information only by nurse condition. The results suggest that references to feelings of SPI such as feeling good and being proud and that giving plasma is a rewarding personal experience favor a higher frequency of plasma donation. PMID:25530909

  13. Development of an immunoassay to detect benzene adducts in hemoglobin

    SciTech Connect

    Grassman, J.A.

    1993-01-01

    The purpose of this project was to develop an immunoassay to detect the adducts formed in hemoglobin after exposure to benzene, which is known to cause bone marrow degeneration and acute myelogenous leukemia. The use of benzene-adduct detection as a biological monitoring method would permit measurement of low exposures and exposures sustained weeks earlier. The reactivity of hydroquinone, an important benzene metabolite, with blood proteins and amino acids was investigated in order to decide which antigens and analytes were likely to be suitable for immunoassay development. The second section determined the combination of benzene-metabolite and antigen need to produce an immunoassay with the requisite low detection limit and specificity. The immunoassays with the best performance were tested on hemoglobin from benzene-exposed mice. In vitro studies showed that hydroquinone efficiently formed adducts with erythrocyte membranes and hemoglobin but not with albumin. Adduction efficiency was greater in incubations using purified hemoglobin than whole blood. Cysteine accounted for 15 to 27% of the adducts formed by hydroquinone. The site of the other adducts were not identified although there was evidence that the hemoglobin heme was adducted. Adducts were found on only 1 of the 2 globin chains. Tryptic digestion of the globin failed to associate the adducts with a specific peptide. Antigens made from hydroquinone-adducted hemoglobin but not hydroquinone-adducted cysteines coupled to carrier proteins effectively elicited adduct-specific antibodies. Interference due to reactivity to hemoglobin was controlled by using uniform quantities of hemoglobin in all wells. The mid-range of the best assays were approximately 12 pmoles HQ per well. Antibodies directed toward hemoglobin adducted with the benzene metabolites phenol, catechol and 1,2,4-trihydroxybenzene were also made. The performance of the anti-1,2,4-trihydroxybenzene were suitable for quantitative immunoassays.

  14. Motivations for Giving of Alumni Donors, Lapsed Donors and Non-Donors: Implications for Christian Higher Education

    ERIC Educational Resources Information Center

    Rugano, Emilio Kariuki

    2011-01-01

    This descriptive and causal comparative study sought to identify motivations for alumni donor acquisition and retention in Christian institutions of higher learning. To meet this objective, motivations for alumni donors, lapsed donors, and non-donors were analyzed and compared. Data was collected through an electronic survey of a stratified sample

  15. Being a Living Donor: Risks

    MedlinePLUS

    ... for blood transfusions side effects associated with allergic reactions to the anesthesia death The best source of information about risks and expected donor outcomes is your transplant team. In addition, it抯 important to take an active role in ...

  16. Why Minority Donors Are Needed

    MedlinePLUS

    ... Why Donate RELATED INFORMATION Minority Focused Grantee Publications Organ Donation Process Enrolling as a Donor Trying to Save ... transplantation for everyone. > More about African Americans and organ donation > More about Asians, and Native Hawaiians and other ...

  17. Donor states in inverse opals

    SciTech Connect

    Mahan, G. D.

    2014-09-21

    We calculate the binding energy of an electron bound to a donor in a semiconductor inverse opal. Inverse opals have two kinds of cavities, which we call octahedral and tetrahedral, according to their group symmetry. We put the donor in the center of each of these two cavities and obtain the binding energy. The binding energies become very large when the inverse opal is made from templates with small spheres. For spheres less than 50 nm in diameter, the donor binding can increase to several times its unconfined value. Then electrons become tightly bound to the donor and are unlikely to be thermally activated to the semiconductor conduction band. This conclusion suggests that inverse opals will be poor conductors.

  18. Donor states in inverse opals

    NASA Astrophysics Data System (ADS)

    Mahan, G. D.

    2014-09-01

    We calculate the binding energy of an electron bound to a donor in a semiconductor inverse opal. Inverse opals have two kinds of cavities, which we call octahedral and tetrahedral, according to their group symmetry. We put the donor in the center of each of these two cavities and obtain the binding energy. The binding energies become very large when the inverse opal is made from templates with small spheres. For spheres less than 50 nm in diameter, the donor binding can increase to several times its unconfined value. Then electrons become tightly bound to the donor and are unlikely to be thermally activated to the semiconductor conduction band. This conclusion suggests that inverse opals will be poor conductors.

  19. Donor selection in heart transplantation

    PubMed Central

    Emani, Sitaramesh; Sai-Sudhakar, Chittoor B.; Higgins, Robert S. D.; Whitson, Bryan A.

    2014-01-01

    There is increased scrutiny on the quality in health care with particular emphasis on institutional heart transplant survival outcomes. An important aspect of successful transplantation is appropriate donor selection. We review the current guidelines as well as areas of controversy in the selection of appropriate hearts as donor organs to ensure optimal outcomes. This decision is paramount to the success of a transplant program as well as recipient survival and graft function post-transplant. PMID:25132976

  20. Mass transfer from giant donors

    NASA Astrophysics Data System (ADS)

    Pavlovskii, K.; Ivanova, N.

    2015-06-01

    The stability of mass transfer in binaries with convective giant donors remains an open question in modern astrophysics. There is a significant discrepancy between what the existing methods predict for a response to mass-loss of the giant itself, as well as for the mass-transfer rate during the Roche lobe overflow. Here we show that the recombination energy in the superadiabatic layer plays an important and hitherto unaccounted-for role in the donor's response to mass-loss, in particular on its luminosity and effective temperature. Our improved optically thick nozzle method to calculate the mass-transfer rate via L1 allows us to evolve binary systems for a substantial Roche lobe overflow. We propose a new, strengthened criterion for the mass-transfer instability, basing it on whether the donor experiences overflow through its outer Lagrangian point. We find that with the new criterion, if the donor has a well-developed outer convective envelope, the critical initial mass ratio for which a binary would evolve stably through the conservative mass transfer varies from 1.5 to 2.2, which is about twice as large as previously believed. In underdeveloped giants with shallow convective envelopes, this critical ratio may be even larger. When the convective envelope is still growing, and in particular for most cases of massive donors, the critical mass ratio gradually decreases to this value, from that of radiative donors.

  1. Philanthropic Motivations of Community College Donors

    ERIC Educational Resources Information Center

    Carter, Linnie S.; Duggan, Molly H.

    2011-01-01

    This descriptive study surveyed current, lapsed, and major gift donors to explore the impact of college communications on donors' decisions to contribute to the college, the likelihood of donor financial support for various college projects, and the philanthropic motivation profiles of the donors of a midsized, multicampus community college in

  2. Rapid, automated, parallel quantitative immunoassays using highly integrated microfluidics and AlphaLISA

    PubMed Central

    Tak For Yu, Zeta; Guan, Huijiao; Ki Cheung, Mei; McHugh, Walker M.; Cornell, Timothy T.; Shanley, Thomas P.; Kurabayashi, Katsuo; Fu, Jianping

    2015-01-01

    Immunoassays represent one of the most popular analytical methods for detection and quantification of biomolecules. However, conventional immunoassays such as ELISA and flow cytometry, even though providing high sensitivity and specificity and multiplexing capability, can be labor-intensive and prone to human error, making them unsuitable for standardized clinical diagnoses. Using a commercialized no-wash, homogeneous immunoassay technology (慉lphaLISA) in conjunction with integrated microfluidics, herein we developed a microfluidic immunoassay chip capable of rapid, automated, parallel immunoassays of microliter quantities of samples. Operation of the microfluidic immunoassay chip entailed rapid mixing and conjugation of AlphaLISA components with target analytes before quantitative imaging for analyte detections in up to eight samples simultaneously. Aspects such as fluid handling and operation, surface passivation, imaging uniformity, and detection sensitivity of the microfluidic immunoassay chip using AlphaLISA were investigated. The microfluidic immunoassay chip could detect one target analyte simultaneously for up to eight samples in 45?min with a limit of detection down to 10?pg mL?1. The microfluidic immunoassay chip was further utilized for functional immunophenotyping to examine cytokine secretion from human immune cells stimulated ex vivo. Together, the microfluidic immunoassay chip provides a promising high-throughput, high-content platform for rapid, automated, parallel quantitative immunosensing applications. PMID:26074253

  3. Rapid, automated, parallel quantitative immunoassays using highly integrated microfluidics and AlphaLISA

    NASA Astrophysics Data System (ADS)

    TakYu, Zeta; Guan, Huijiao; Ki Cheung, Mei; McHugh, Walker M.; Cornell, Timothy T.; Shanley, Thomas P.; Kurabayashi, Katsuo; Fu, Jianping

    2015-06-01

    Immunoassays represent one of the most popular analytical methods for detection and quantification of biomolecules. However, conventional immunoassays such as ELISA and flow cytometry, even though providing high sensitivity and specificity and multiplexing capability, can be labor-intensive and prone to human error, making them unsuitable for standardized clinical diagnoses. Using a commercialized no-wash, homogeneous immunoassay technology (慉lphaLISA) in conjunction with integrated microfluidics, herein we developed a microfluidic immunoassay chip capable of rapid, automated, parallel immunoassays of microliter quantities of samples. Operation of the microfluidic immunoassay chip entailed rapid mixing and conjugation of AlphaLISA components with target analytes before quantitative imaging for analyte detections in up to eight samples simultaneously. Aspects such as fluid handling and operation, surface passivation, imaging uniformity, and detection sensitivity of the microfluidic immunoassay chip using AlphaLISA were investigated. The microfluidic immunoassay chip could detect one target analyte simultaneously for up to eight samples in 45?min with a limit of detection down to 10?pg mL-1. The microfluidic immunoassay chip was further utilized for functional immunophenotyping to examine cytokine secretion from human immune cells stimulated ex vivo. Together, the microfluidic immunoassay chip provides a promising high-throughput, high-content platform for rapid, automated, parallel quantitative immunosensing applications.

  4. Who is the best alternative allotransplant donor?

    PubMed Central

    Gale, RP; Eapen, M

    2015-01-01

    Assuming that most physicians will chose an HLA-identical sibling as the best allotransplant donor, the question arises who is the best alternative donor when an HLA-identical sibling is unavailable? The most commonly used alternative donors are HLA-identical or -mismatched unrelated donors, HLA-matched or -mismatched umbilical cord blood donor or a related, HLA-haplotype-matched related donors. Each alternative donor option has advantages and disadvantages. We discuss selected aspects of these issues based on data from randomized clinical trials and observational databases. However, because there are limited data to address specific clinical settings, quantification of expert opinion is sometimes needed. PMID:26039206

  5. Synthesis of optically pure chrysobactin and immunoassay development.

    PubMed

    Lu, C; Buyer, J S; Okonya, J F; Miller, M J

    1996-10-01

    Chrysobactin (alpha-N-(2,3-dihydroxybenzoyl)-D-lysyl-L-serine), a siderophore that is essential for systemic virulence by plant pathogenic Erwinia chrysanthemi, was synthesized with high diastereomeric purity. Chrysobactin was prepared by coupling the N-hydroxysuccinimide ester of alpha-N-(2,3-dibenzyloxybenzoyl)-epsilon-N-Cbz-D-lysine with L-serine benzyl ester followed by deprotection via hydrogenolysis. Optically pure chrysobactin was obtained with 98% overall yield. A monoclonal antibody to ferric chrysobactin was developed and characterized as IgM. The antibody reacts with chrysobactin, ferric chrysobactin and less strongly with ferric dihydroxybenzoic acid. The antibody reacts weakly with the siderophores ferrichrome, A, ferric pseudobactin and ferric rhodotorulic acid. This antibody was used in a competitive immunoassay to detect ferric chrysobactin at 10(-8) to 10(-10) mol. This immunoassay may provide a useful method for the detection of chrysobactin in plant samples. PMID:8837459

  6. Silver and gold enhancement methods for lateral flow immunoassays.

    PubMed

    Rodr韌uez, Myriam Oliveira; Covi醤, Luc韆 Blanco; Garc韆, Agust韓 Costa; Blanco-L髉ez, Maria Carmen

    2016-02-01

    Sensitivity is the main concern at the development of rapid test by lateral flow immunoassays. On the other hand, low limits of detection are often required at medical diagnostics and other field of analysis. To overcome this drawback, several enhancement protocols have been described. In this paper, we have selected different silver enhancement methods and one dual gold conjugation, and we critically compared the amplification produced when applied to a gold-nanoparticle based lateral flow immunoassay for the detection of prostate specific antigen (PSA). The highest amplification was obtained by using an immersion method based on a solution of silver nitrate and hydroquinone/citrate buffer in proportion 1:1. Under these conditions, the system is capable of detecting PSA within 20min at levels as low as 0.1ng/mL, with a 3-fold sensitivity improvement. PMID:26653449

  7. Benzene-RISc: The development and performance of an immunoassay to detect benzene in water

    SciTech Connect

    Friedman, S.B.; Withers, T.; Almond, R.; Stewart, T.; Allen, R.L.

    1994-12-31

    Immunoassay methods have become available for environmental applications. Their simplicity, reliability, and ability to provide information rapidly and on-site is enhancing the efficiency of many field and laboratory programs. Immunoassay methods rely upon antibody molecules to provide the sensitivity and specificity characteristics they exhibit, but many molecules are either insufficiently immunogenic or structurally unremarkable to induce an appropriate antibody response. Such compounds are usually considered to be incompatible with the development of an immunoassay method. An immunoassay method for the detection of benzene in water would have utility in detecting contamination from spills and leaking underground storage tanks. Benzene, however, is frequently considered to be in the class of compounds considered to be incompatible with antibody, and therefore immunoassay, development. The authors have developed an immunoassay for the detection of benzene in water by developing both sample processing and immunochemical procedures and reagents that overcome the technical limitations frequently encountered.

  8. Cooperative Immunoassays: Ultrasensitive Assays with Mixed Monoclonal Antibodies

    NASA Astrophysics Data System (ADS)

    Ehrlich, Paul H.; Moyle, William R.

    1983-07-01

    Mixtures of certain monoclonal antibodies appear to bind human chorionic gonadotropin in a ``cooperative'' fashion because they form circular complexes with the hormone. Experiments illustrate how this property might be exploited to develop very sensitive immunoassays for human chorionic gonadotropin or any other antigen. Since the assays are not based on competitive inhibition between radiolabeled and unlabeled antigen, they are much more sensitive than a traditional radioimmunoassay in which either one of the same antibodies is used alone.

  9. PCB detection by immunoassay -- A wipe test for surface contamination

    SciTech Connect

    Dautlick, J.X.; Teaney, G.B.; Hudak, R.T.; Melby, J.M.

    1995-12-31

    Immunoassay based field screening methods are gaining acceptance by the environmental diagnostics industry for on-site characterization and remediation monitoring. Polychlorinated biphenyls (PCBs), a family of molecules classified as potential carcinogens, can be easily detected on-site by immunoassay screening methods. This results in reduced project cost and improved onsite efficiency, since field screening immunoassays short cut the long turn around time of laboratory analysis while providing reliable results. On site wipe test technology for assessing PCB contamination on surfaces such as walls and floors of PCB storage facilities has been developed to supplement the D TECH{trademark} PCB soil assay. This sampling technique can also be used to monitor for transformer leaks, spills and to evaluate equipment decontamination processes. The D TECH PCB wipe test is quick, cost effective, highly specific and user friendly. The surface is sampled by wiping a 100 cm{sup 2} area with a 1 cm{sup 2} pad saturated with an extractant. The PCB is extracted from the sampling pad during a short extraction step. The sample is filtered, diluted, and run in the D TECH PCB field screening system. The components of the immunoassay include PCB specific antibodies (Ab) covalently linked to small latex particles, a PCB analog which is covalently linked to alkaline phosphatase, and the free PCB from the sample. The free PCB competes with the enzyme linked analog for the Ab binding sites. The latex particles are then collected on a filter device, washed, and an enzyme substrate is added. The amount of color produced is inversely proportional to the concentration of free PCB on the sample, and can be determined using a hand held reflectometer, or a color card.

  10. Rapid determination of dioxin in water by enzyme immunoassay

    SciTech Connect

    Wang, H.; Wang, L.; George J.E. III; Ward, G.K.

    1996-10-01

    Dioxin in water, soil, sediments and other sample matrices is usually determined by the EPA method 1613 which was developed by the EPA Office of Science and Technology. This method however requires expensive instruments (high resolution gas chromatography/high resolution mass spectrometry) and a highly trained analyst. In order to reduce the cost and turn around time, a dioxin enzyme immunoassay (EIA) was developed to rapidly analyze trace levels of 2,3,7,8-tetra chlorinated dibenzodioxin (TCDD) in water samples. Water samples were extracted using a 47 mm, C18 Empore extraction disk (3M). Dioxin was eluted with dichloromethame. EnvironGard reagents and microwell strip reader (Millipore Corporation) were used to perform the dioxin enzyme immunoassay. The extraction efficiency was also tested by GC/MS with Varian`s large volume injector and Selected Ion Storage technique. The working range of the dioxin enzyme immunoassay was from 15 pg/L to 100 pg/L. The precision and accuracy of EIA was determined by performing five replicates of reagent water spiked at a concentration of 25 pg/L. The recovery of the dioxin assay ranged from 74% to 122%, and % CV for five replicates was less than 15%. In general, EIA provides a relatively easy and cost effective means for measuring trace levels of dioxin in water samples.

  11. Multiplex Electrochemical Immunoassay Using Gold Nanoparticle Probes and Immunochromatographic Strips

    SciTech Connect

    Mao, Xun; Baloda, Meenu; Gurung, Anant; Lin, Yuehe; Liu, Guodong

    2008-10-20

    We describe a multiplex electrochemical immunoassay based on the use of gold nanoparticle (Au-NP) probes and immunochromatographic strips (ISs). The approach takes advantage of the speed and low cost of the conventional IS tests and the high sensitivities of the nanoparticle-based electrochemical immunoassays. Rabbit IgG(R-IgG) and human IgM (H-IgM) were used as model targets for the demonstration of the proof of concept. The Au-NPs based sandwich immunoreactions were performed on the IS, and the captured gold nanoparticle labels on the test zones were determined by highly-sensitive stripping voltammetric measurement of the dissolved gold ions (III) with a carbon paste electrode. The detection limits are 1.0 and 1.5 ng/mL with the linear ranges of 2.5-250 ng/mL for quantitative detection of R-IgG and H-IgM, respectively. The total assay time is around 25 minutes. Such multiplex electrochemical immunoassay could be readily highly multiplexed to allow simultaneous parallel detection of numerous proteins and is expected to open new opportunities for protein diagnostics and biosecurity.

  12. Simple Patterned Nanofiber Scaffolds and Its Enhanced Performance in Immunoassay

    PubMed Central

    Lv, Xiao-guang; Song, Jia; Zhan, Na; Dong, Wei-guo; Huang, Wei-hua

    2013-01-01

    Cancer has become the leading cause of death worldwide; early diagnosis and treatment of cancers is critical for the survival of the patients. The concentration of cancer markers in easy-to-access biological fluids can provide great assistance in screening for occult primary cancers, distinguishing malignant from benign findings, determining prognosis and prediction for cancer patients. The multiplex detection technology of a panel of cancer markers can greatly increase the accuracy of disease diagnosis. Herein, we briefly fabricate a high-throughput micro-immunoassay based on the electrospun polystyrene (PS) substrates to improve detection sensitivity. The immunoassay was evaluated by analyzing three different cancer biomarkers (AFP, CEA, VEGF). For AFP, CEA, VEGF immunofluorescence assay, the LOD of assay conducted on electrospun PS substrates before or after plasma and the conventional PS substrates were 0.42, 0.10, 1.12 ng/mL, 0.57, 0.09, 1.24 ng/mL, and 159.75, 26.19, 385.59 pg/mL, respectively (P < 0.05). Due to the high porosity and large surface area-to-volume ratio which is the foremost merit of nanostructures, and the plasma treatment which make the hydrophobic PS nanofibers hydropholic, the nanofibers substrates showed sufficient retention of immunoassay functionality and high potential for capture molecules immobilization. Consequently, the immunofluorescence assay conducted on electrospun PS substrates could significantly enhance the sensitivity and limits of detection. PMID:24340065

  13. Enzyme immunoassay for nomegestrol acetate in human plasma.

    PubMed

    Ezan, E; Benech, H; Bucourt, R; Ardouin, T; Tchernatinsky, C; Thomas, J L; Paris, J; Grognet, J M

    1993-10-01

    Currently available chromatographic assays of the progestative drug nomegestrol acetate in human plasma are not suitable for monitoring drug kinetics more than 24 h after clinical dosage. A specific and sensitive enzyme immunoassay was therefore developed. A 3(O-carboxymethyl)oxime derivative of nomegestrol acetate was synthesized and coupled to bovine serum albumin in order to raise polyclonal antibodies in rabbits. The enzymatic tracer was obtained by coupling the 3(O-carboxymethyl)oxime derivative to acetylcholinesterase (E.C.3.1.1.7.). HPLC fractionation of human plasma samples followed by enzyme immunoassay revealed the presence of cross-reacting metabolites. An automated procedure of metabolite separation was developed using silica bonded with diol groups (Diol Bakerbond column). This procedure ensured assay specificity. The quantification limit in human plasma was 0.1 ng/ml. Mean repeatability (intra-assay variation) and reproducibility (inter-assay variation) were 9 and 15%, respectively. The enzyme immunoassay allowed monitoring of the kinetics of nomegestrol acetate 144 h after oral administration of a single 5 mg dose. Values for human samples were in excellent agreement with those assayable by HPLC followed by u.v. detection. PMID:8217881

  14. 75 FR 58400 - Donor Management Research: Improvements in Clinical Management of Deceased Organ Donors

    Federal Register 2010, 2011, 2012, 2013, 2014

    2010-09-24

    ...Management Research: Improvements in Clinical Management of Deceased Organ Donors AGENCY...contract that focuses on improvements in clinical management of deceased organ donors...It is reasonable to expect that better clinical donor management would improve...

  15. Multicenter Analytical Evaluation of the Automated Electrochemiluminescence Immunoassay for Cyclosporine

    PubMed Central

    Vogeser, Michael; Shipkova, Maria; Rigo-Bonnin, Ra黮; Wallemacq, Pierre; Orth, Matthias; Widmann, Monika

    2014-01-01

    Background: Cyclosporine A (CsA) is used as a posttransplantation immunosuppressant drug, and careful monitoring of CsA concentration in whole blood is essential. A new automated electrochemiluminescence immunoassay (ECLIA) for CsA measurement has been assessed in a multicenter evaluation. Methods: Residual EDTA whole blood samples from patients undergoing CsA therapy after organ transplant were used in assay evaluation at 5 clinical laboratories in Europe. Experiments included imprecision according to CLSI EP5-A2 (within-run and intermediate), lower limit of quantification, linearity according to CLSI EP6-A, and recovery of commercial external quality control samples. In addition, comparisons to liquid chromatography-tandem mass spectrometry methods in routine use at each investigational site and to commercial chemiluminescent microparticle immunoassay and antibody-conjugated magnetic immunoassay methods were performed. Results: Imprecision testing gave coefficients of variation of less than 9% in the 302000 mcg/L range for both within-run and intermediate imprecision. Lower limit of quantification of 6.8 mcg/L at one investigational site and 1.8 mcg/L at a second site at 20% coefficient of variation were observed. Linearity was measured over the concentration range 02000 mcg/L, yielding a deviation of less than 12%. External quality control sample recovery by ECLIA was 93%114% of LC-MS/MS sample recovery. Deming regression analysis of ECLIA method comparison to combined LC-MS/MS results yielded a slope of 1.04 [95% confidence interval (CI), 1.031.06] and intercept of 2.8 mcg/L (95% CI, 1.54.1 mcg/L). Comparison to chemiluminescent microparticle immunoassay yielded a slope of 0.87 (95% CI, 0.850.89) and intercept of 1.4 mcg/L (95% CI, ?0.89 to 3.7 mcg/L); comparison to antibody-conjugated magnetic immunoassay yielded a slope of 0.96 (95% CI, 0.930.98) and intercept of ?4.2 mcg/L (95% CI, ?7.1 to ?1.2 mcg/L). Conclusions: The data from this multicenter evaluation indicate that the new ECLIA-based cyclosporine assay is fit for its purpose, the therapeutic monitoring of CsA. PMID:24646730

  16. Development of a sensitive microarray immunoassay and comparison with standard enzyme-linked immunoassay for cytokine analysis.

    PubMed

    Knight, Paul R; Sreekumar, Arun; Siddiqui, Javed; Laxman, Bharathi; Copeland, Shannon; Chinnaiyan, Arul; Remick, Daniel G

    2004-01-01

    Cytokine and cytokine inhibitors represent important components of the inflammatory response in patients with trauma, shock, and sepsis. Many investigators wish to quantify cytokines and it would be advantageous to measure multiple cytokines in a multiplex manner to obtain an inflammatory profile rather than a single value. Using the well-accepted standard enzyme-linked immunoassay (ELISA) as a basis, a microarray immunoassay (MI) was designed to measure 16 different human cytokines simultaneously. The MI was performed by spotting antibodies on nitrocellulose pads affixed to glass slides. Detection of the mediators was performed with biotin-conjugated antibodies followed by fluorescently labeled streptavidin. All antibodies and other reagents were purchased commercially. The MI achieved a lower limit of detection that was generally similar to traditional ELISAs (approximately 4-12 pg/mL) and also had a similar coefficient of variation. In the multiplexed MI, there was no cross reactivity between mediators. To verify the utility of the MI, cytokines and cytokine inhibitors were measured in endotoxin stimulated human blood by both ELISA and MI. Virtually identical cytokine concentrations were measured by both methods. These results describe the development of a sensitive, specific and cost-effective multiplexed microarray immunoassay that produces values similar to traditional ELISAs. PMID:14676680

  17. Single-Donor Leukophoretic Technique

    NASA Technical Reports Server (NTRS)

    Eberhardt, R. N.

    1977-01-01

    Leukocyte separation-and-retrieval device utilizes granulocyte and monocyte property of leukoadhesion to glass surfaces as basis of their separation from whole blood. Device is used with single donor technique and has application in biological and chemical processing, veterinary research and clinical care.

  18. Microchip device with 64-site electrode array for multiplexed immunoassay of cell surface antigens based on electrochemiluminescence resonance energy transfer.

    PubMed

    Wu, Mei-Sheng; Shi, Hai-Wei; He, Li-Jing; Xu, Jing-Juan; Chen, Hong-Yuan

    2012-05-01

    This paper describes a novel on-chip microarray platform based on an electrochemiluminescence resonance energy transfer (ECL-RET) strategy for rapid assay of cancer cell surface biomarkers. This platform consists of 64 antigen-decorated CdS nanorod spots with the diameter of 1.0 cm uniformly distributed on 16 indium tin oxide (ITO) strips, which is coated with a multichannel decorated polydimethylsiloxane (PDMS) slice to realize multiplexed determination of antigens. To shorten the immune reaction time in the microchannels and simplify the device, magnetic stirring and four-channel universal serial bus (USB) ports for plug-and-play were used. When Ru(bpy)(3)(2+) labeled antibodies were selectively captured by the corresponding antigens on the CdS nanorod spot array, ECL-RET from the CdS nanorod (donor) by cathodic emission in the presence of K(2)S(2)O(8) to Ru(bpy)(3)(2+) (acceptor) occurred. With signal amplification of Ru(bpy)(3)(2+) and competitive immunoassay, carcinoembryonic antigen (CEA), ?-fetoprotein (AFP), and prostate specific antigen (PSA) as models were detected on this microfluidic device via recording the increased ECL-RET signals on electrode surfaces. Furthermore, this multiplexed competitive immunoassay was successfully used for detecting cancer cell surface antigens via the specific antibody-cell interactions and cell counting via cell surface receptors and antigens on the CdS nanorod surface. This platform provides a rapid and simple but sensitive approach with microliter-level sample volume and holds great promise for multiplexed detection of antigens and antigen-specific cells. PMID:22494075

  19. Micromotor-based lab-on-chip immunoassays

    NASA Astrophysics Data System (ADS)

    Garc韆, Miguel; Orozco, Jahir; Guix, Maria; Gao, Wei; Sattayasamitsathit, Sirilak; Escarpa, Alberto; Merko鏸, Arben; Wang, Joseph

    2013-01-01

    Here we describe the first example of using self-propelled antibody-functionalized synthetic catalytic microengines for capturing and transporting target proteins between the different reservoirs of a lab-on-a-chip (LOC) device. A new catalytic polymer/Ni/Pt microtube engine, containing carboxy moieties on its mixed poly(3,4-ethylenedioxythiophene) (PEDOT)/COOH-PEDOT polymeric outermost layer, is further functionalized with the antibody receptor to selectively recognize and capture the target protein. The new motor-based microchip immunoassay operations are carried out without any bulk fluid flow, replacing the common washing steps in antibody-based protein bioassays with the active transport of the captured protein throughout the different reservoirs, where each step of the immunoassay takes place. A first microchip format involving an `on-the-fly' double-antibody sandwich assay (DASA) is used for demonstrating the selective capture of the target protein, in the presence of excess of non-target proteins. A secondary antibody tagged with a polymeric-sphere tracer allows the direct visualization of the binding events. In a second approach the immuno-nanomotor captures and transports the microsphere-tagged antigen through a microchannel network. An anti-protein-A modified microengine is finally used to demonstrate the selective capture, transport and convenient label-free optical detection of a Staphylococcus aureus target bacteria (containing proteinA in its cell wall) in the presence of a large excess of non-target (Saccharomyces cerevisiae) cells. The resulting nanomotor-based microchip immunoassay offers considerable potential for diverse applications in clinical diagnostics, environmental and security monitoring fields.Here we describe the first example of using self-propelled antibody-functionalized synthetic catalytic microengines for capturing and transporting target proteins between the different reservoirs of a lab-on-a-chip (LOC) device. A new catalytic polymer/Ni/Pt microtube engine, containing carboxy moieties on its mixed poly(3,4-ethylenedioxythiophene) (PEDOT)/COOH-PEDOT polymeric outermost layer, is further functionalized with the antibody receptor to selectively recognize and capture the target protein. The new motor-based microchip immunoassay operations are carried out without any bulk fluid flow, replacing the common washing steps in antibody-based protein bioassays with the active transport of the captured protein throughout the different reservoirs, where each step of the immunoassay takes place. A first microchip format involving an `on-the-fly' double-antibody sandwich assay (DASA) is used for demonstrating the selective capture of the target protein, in the presence of excess of non-target proteins. A secondary antibody tagged with a polymeric-sphere tracer allows the direct visualization of the binding events. In a second approach the immuno-nanomotor captures and transports the microsphere-tagged antigen through a microchannel network. An anti-protein-A modified microengine is finally used to demonstrate the selective capture, transport and convenient label-free optical detection of a Staphylococcus aureus target bacteria (containing proteinA in its cell wall) in the presence of a large excess of non-target (Saccharomyces cerevisiae) cells. The resulting nanomotor-based microchip immunoassay offers considerable potential for diverse applications in clinical diagnostics, environmental and security monitoring fields. Electronic supplementary information (ESI) available. See DOI: 10.1039/c2nr32400h

  20. Evaluation of enzyme immunoassay (Chlamydiazyme) for detecting Chlamydia trachomatis in genital tract specimens.

    PubMed Central

    Taylor-Robinson, D; Thomas, B J; Osborn, M F

    1987-01-01

    An enzyme immunoassay (Chlamydiazyme) for detecting Chlamydia trachomatis was evaluated on genital specimens from 96 men and 272 women attending a clinic for sexually transmitted diseases (STD clinic). Compared with a direct immunofluorescence test for chlamydial elementary bodies, the enzyme immunoassay had a sensitivity of 58% on specimens from men, a specificity of 90%, a positive predictive value of 93%, and a negative predictive value of 88%; the assay had a sensitivity of 67% on specimens from women, a specificity of 89%, a positive predictive value of 63% and a negative predictive value of 90%. Immunofluorescence provided the most stringent test for the performance of the enzyme immunoassay as values were improved a little when a cell culture procedure was used for comparison. Further evidence for the lack of sensitivity was the detection of elementary bodies, sometimes in large numbers, in the enzyme immunoassay buffer of 13 of 19 specimens that had given a negative enzyme immunoassay result and the finding in comparative titrations of four laboratory strains that the enzyme immunoassay was at least 100-fold less able to detect chlamydiae than either immunofluorescence or the cell culture procedure. Lack of specificity may be associated with the finding that the enzyme immunoassay antibody reacted with strains of Acinetobacter calcoaceticus, Escherichia coli, Gardnerella vaginalis, Neisseria gonorrhoeae and group B streptococci. The enzyme immunoassay was not considered to be sufficiently sensitive, specific, or reproducible for routine use. PMID:3546397

  1. ULTRASENSITIVE IMMUNOASSAYS BASED ON SURFACE-ENHANCED RAMAN SCATTERING BY IMMUNOGOLD LABELS

    Technology Transfer Automated Retrieval System (TEKTRAN)

    This chapter describes recent advances in the use of surface-enhanced Raman Scattering (SERS) as a readout tool for chip-scale, sandwich-based immunoassays. It reviews progress made in developing SERS-based immunoassays for a wide range of biolytes, including proteins, viruses and bacteria. The st...

  2. Synthesis of Protein Conjugates and Development of Immunoassays for AAL Toxins

    E-print Network

    Hammock, Bruce D.

    Synthesis of Protein Conjugates and Development of Immunoassays for AAL Toxins Ferenc Szurdoki AAL toxins and fumonisins, produced by Alternaria alternata f. sp. lycopersici and Fusarium sensitive, inexpensive, and rapid immunoassays are needed to quantify these natural toxins in food products

  3. DEVELOPMENT OF A CLASS-SELECTIVE ENZYME IMMUNOASSAY FOR URINARY PHENOLIC GLUCURONIDES. (R825433)

    EPA Science Inventory

    Class-selective immunoassays for the measurement of glucuronides in human urine can aid evaluation of human exposure to complex mixtures of xenobiotics. Therefore, an enzyme immunoassay (EIA) for the group-selective detection of phenolic On-Chip Native Gel Electrophoresis-Based Immunoassays for Tetanus Antibody and Toxin

    E-print Network

    Herr, Amy E.

    On-Chip Native Gel Electrophoresis-Based Immunoassays for Tetanus Antibody and Toxin Amy E. Herr was developed for detection of tetanus antibodies in buffer as well as diluted serum samples. After an off was 0.68 nM. A competitive immunoassay was also developed for tetanus toxin C-fragment by allowing un

  4. Europium chelate labels in time-resolved fluorescence immunoassays and DNA hybridization assays

    SciTech Connect

    Diamandis, E.P.; Christopoulos, T.K. Univ. of Toronto, Ontario )

    1990-11-15

    Like many analytical methodologies, immunoassays and nucleic acid hybridization assays rely on the reaction between an analyte of interest and a specific reagent. The analyte concentration is then deduced by measuring either the amount of analyte-reagent complex formed (product) or the amount of residual reagent. The authors describe the application of fluorescent rare-earth chelates to immunoassay and DNA probing.

  5. Kinetics of protein binding in solid-phase immunoassays: Theory Konstantin V. Klenin

    E-print Network

    Langowski, J鰎g

    Kinetics of protein binding in solid-phase immunoassays: Theory Konstantin V. Klenin Division 2005; accepted 13 April 2005; published online 7 June 2005 In a solid-phase immunoassay, binding in solution. We present a theory describing the kinetics of immunochemical reaction in such a system. A single

  6. Live donor transplantation--the incompetent donor: comparative law.

    PubMed

    Wolfman, Samuel; Shaked, Tali

    2008-12-01

    Informed consent of the patient to medical treatment is an essential prerequisite for any invasive medical procedure. However in emergency cases, when the patient is unable to sign a consent form due to unconsciousness or to psychotic state, than the primary medical consideration shall take place. In such a case, in order to save life or even prevent a major medical hazard to the patient, doctors are allowed, in certain cases and in accordance with well accepted medical practice, to perform invasive procedures, major surgery or risky pharmacological treatment, without the explicit consent of the patient. All the above refers to the cases when avoidance of such non-consented treatment may harm severely the health and wellbeing of the patient and there is no doubt that such treatment is for the ultimate benefit of the patient. The question, however, shall arise when such a medical procedure is not necessarily for the benefit of the patient, but rather for the benefit of somebody else. Such is the case in the transplantation area and the question of living donor-donee relationship. This paper shall analyze the legal situation in cases of non competent donors whose consent cannot be considered legal consent given in full understanding and out of free will. It will also compare three legal systems, the Israeli, the American and the traditional Jewish law, with regard to the different approaches to this human problem, where the autonomy of the donor may be sacrificed for the purpose of saving life of another person. PMID:19202861

  7. Fast and sensitive detection of enteropathogenic Yersinia by immunoassays.

    PubMed

    Laporte, J閞鬽e; Savin, Cyril; Lamourette, Patricia; Devilliers, Karine; Volland, Herv; Carniel, Elisabeth; Cr閙inon, Christophe; Simon, St閜hanie

    2015-01-01

    Yersinia enterocolitica and Yersinia pseudotuberculosis, the two Yersinia species that are enteropathogenic for humans, are distributed worldwide and frequently cause diarrhea in inhabitants of temperate and cold countries. Y. enterocolitica is a major cause of foodborne disease resulting from consumption of contaminated pork meat and is further associated with substantial economic cost. However, investigation of enteropathogenic Yersinia species is infrequently performed routinely in clinical laboratories because of their specific growth characteristics, which make difficult their isolation from stool samples. Moreover, current isolation procedures are time-consuming and expensive, thus leading to underestimates of the incidence of enteric yersiniosis, inappropriate prescriptions of antibiotic treatments, and unnecessary appendectomies. The main objective of the study was to develop fast, sensitive, specific, and easy-to-use immunoassays, useful for both human and veterinary diagnosis. Monoclonal antibodies (MAbs) directed against Y. enterocolitica bioserotypes 2/O:9 and 4/O:3 and Y. pseudotuberculosis serotypes I and III were produced. Pairs of MAbs were selected by testing their specificity and affinity for enteropathogenic Yersinia and other commonly found enterobacteria. Pairs of MAbs were selected to develop highly sensitive enzyme immunoassays (EIAs) and lateral flow immunoassays (LFIs or dipsticks) convenient for the purpose of rapid diagnosis. The limit of detection of the EIAs ranged from 3.2 10(3) CFU/ml to 8.8 10(4) CFU/ml for pathogenic serotypes I and III of Y. pseudotuberculosis and pathogenic bioserotypes 2/O:9 and 4/O:3 of Y. enterocolitica and for the LFIs ranged from 10(5) CFU/ml to 10(6) CFU/ml. A similar limit of detection was observed for artificially contaminated human feces. PMID:25355759

  8. Fast and Sensitive Detection of Enteropathogenic Yersinia by Immunoassays

    PubMed Central

    Laporte, J閞鬽e; Savin, Cyril; Lamourette, Patricia; Devilliers, Karine; Volland, Herv; Carniel, Elisabeth; Cr閙inon, Christophe

    2014-01-01

    Yersinia enterocolitica and Yersinia pseudotuberculosis, the two Yersinia species that are enteropathogenic for humans, are distributed worldwide and frequently cause diarrhea in inhabitants of temperate and cold countries. Y. enterocolitica is a major cause of foodborne disease resulting from consumption of contaminated pork meat and is further associated with substantial economic cost. However, investigation of enteropathogenic Yersinia species is infrequently performed routinely in clinical laboratories because of their specific growth characteristics, which make difficult their isolation from stool samples. Moreover, current isolation procedures are time-consuming and expensive, thus leading to underestimates of the incidence of enteric yersiniosis, inappropriate prescriptions of antibiotic treatments, and unnecessary appendectomies. The main objective of the study was to develop fast, sensitive, specific, and easy-to-use immunoassays, useful for both human and veterinary diagnosis. Monoclonal antibodies (MAbs) directed against Y. enterocolitica bioserotypes 2/O:9 and 4/O:3 and Y. pseudotuberculosis serotypes I and III were produced. Pairs of MAbs were selected by testing their specificity and affinity for enteropathogenic Yersinia and other commonly found enterobacteria. Pairs of MAbs were selected to develop highly sensitive enzyme immunoassays (EIAs) and lateral flow immunoassays (LFIs or dipsticks) convenient for the purpose of rapid diagnosis. The limit of detection of the EIAs ranged from 3.2 103 CFU/ml to 8.8 104 CFU/ml for pathogenic serotypes I and III of Y. pseudotuberculosis and pathogenic bioserotypes 2/O:9 and 4/O:3 of Y. enterocolitica and for the LFIs ranged from 105 CFU/ml to 106 CFU/ml. A similar limit of detection was observed for artificially contaminated human feces. PMID:25355759

  9. Enzyme immunoassay validation for the detection of buprenorphine in urine.

    PubMed

    Cirimele, V; Kintz, P; Lohner, S; Ludes, B

    2003-03-01

    A solid-phase enzyme immunoassay involving microtiter plates was proposed by Microgenics to screen buprenorphine in urine. The intra-assay precision at 10 ng/mL was 7.7% (coefficient of variation). The immunoassay was determined to have no cross-reactivity with codeine, dihydrocodeine, morphine, ethylmorphine, 6-monoacetylmorphine, methadone, pholcodine, propoxyphene, dextromoramide, and dextromethorphan at 1 and 10 mg/L. A low cross-reactivity (3% at 1 ng/mL) was observed at low concentrations of norbuprenorphine. After comparing this new immunological test (Singlestep ELISA) for 76 urine specimens with our validated high-performance liquid chromatography-electrospray mass spectrometry (HPLC-ES-MS) procedure, an optimum cutoff concentration of 2 ng/mL was determined for the kit. At this cutoff, the screening assay was able to determine more than 90% of true results with 43.4% true positives and 48.7% true negatives. Four positive urines (5.3%) were not confirmed by HPLC-ES-MS. In only one case, the negative urine test was confirmed as positive by HPLC-ES-MS (buprenorphine: 62.5 ng/mL). Buprenorphine concentrations determined by HPLC-ES-MS ranged from 1.2 to 1052 ng/mL. Of the four potential adulterants (hypochloride 50 mL/L, sodium nitrite 50 g/L, liquid soap 50 mL/L, and sodium chloride 50 g/L) that might be added to a positive urine specimen, none were able to cause a false-negative response by the immunoassay. The results of this study support the concept that the Singlestep ELISA for buprenorphine determination in urine should be considered as a new, valided screening procedure. PMID:12670004

  10. AN ENVIRONMENTAL TECHNOLOGY VERIFICATION (ETV) TESTING OF THREE IMMUNOASSAY TEST KITS FOR ANTHRAX, BOTULINUM TOXIN AND RICIN

    EPA Science Inventory

    Immunoassay test kits are based on immunoassay methods, where specific antibodies are used to detect and measure the contaminants of interest. Immunoassay test kits rely on the reaction of a contaminant or antigen with a selective antibody to give a product that can be measures....

  11. Pholcodine interference in the immunoassay for opiates in urine.

    PubMed

    Svenneby, G; Wedege, E; Karlsen, R L

    1983-01-01

    The excretion in urine after single oral therapeutic doses of morphine derivatives was analysed with radioimmunoassay (RIA) and homogeneous enzyme immunoassay (EMIT) for opiates. In contrast to the rapid excretion of ethylmorphine and codeine, pholcodine showed positive results for opiates 2-6 weeks after intake when the urines were analysed with the RIA-method. When analysed with the EMIT-method, positive results were obtained for pholcodine for approximately 10 days. As pholcodine is a common component in cough mixtures, its prolonged excretion could represent a hazard in interpreting the results from drug analyses of urines. PMID:6347841

  12. Demonstration of four immunoassay formats using the array biosensor

    NASA Technical Reports Server (NTRS)

    Sapsford, Kim E.; Charles, Paul T.; Patterson, Charles H Jr; Ligler, Frances S.

    2002-01-01

    The ability of a fluorescence-based array biosensor to measure and quantify the binding of an antigen to an immobilized antibody has been demonstrated using the four different immunoassay formats: direct, competitive, displacement, and sandwich. A patterned array of antibodies specific for 2,4,6-trinitrotoluene (TNT) immobilized onto the surface of a planar waveguide and used to measure signals from different antigen concentrations simultaneously. For direct, competitive, and displacement assays, which are one-step assays, measurements were obtained in real time. Dose-response curves were calculated for all four assay formats, demonstrating the array biosensor's ability to quantify the amount of antigen present in solution.

  13. Changing pattern of donor selection criteria in deceased donor liver transplant: a review of literature.

    PubMed

    Routh, Dronacharya; Naidu, Sudeep; Sharma, Sanjay; Ranjan, Priya; Godara, Rajesh

    2013-12-01

    During the last couple of decades, with standardization and progress in surgical techniques, immunosuppression and post liver transplantation patient care, the outcome of liver transplantation has been optimized. However, the principal limitation of transplantation remains access to an allograft. The number of patients who could derive benefit from liver transplantation markedly exceeds the number of available deceased donors. The large gap between the growing list of patients waiting for liver transplantation and the scarcity of donor organs has fueled efforts to maximize existing donor pool and identify new avenues. This article reviews the changing pattern of donor for liver transplantation using grafts from extended criteria donors (elderly donors, steatotic donors, donors with malignancies, donors with viral hepatitis), donation after cardiac death, use of partial grafts (split liver grafts) and other suboptimal donors (hypernatremia, infections, hypotension and inotropic support). PMID:25755521

  14. Species Specific Bacterial Spore Detection Using Lateral-Flow Immunoassay with DPA-Triggered Tb Luminescence

    NASA Technical Reports Server (NTRS)

    Ponce, Adrian

    2003-01-01

    A method of detecting bacterial spores incorporates (1) A method of lateral-flow immunoassay in combination with (2) A method based on the luminescence of Tb3+ ions to which molecules of dipicolinic acid (DPA) released from the spores have become bound. The present combination of lateral-flow immunoassay and DPA-triggered Tb luminescence was developed as a superior alternative to a prior lateral-flow immunoassay method in which detection involves the visual observation and/or measurement of red light scattered from colloidal gold nanoparticles. The advantage of the present combination method is that it affords both (1) High selectivity for spores of the species of bacteria that one seeks to detect (a characteristic of lateral-flow immunoassay in general) and (2) Detection sensitivity much greater (by virtue of the use of DPA-triggered Tb luminescence instead of gold nanoparticles) than that of the prior lateral-flow immunoassay method

  15. Controversies related to living kidney donors

    PubMed Central

    Kamal, Ahmed I.; Harraz, Ahmed M.; Shokeir, Ahmed A.

    2011-01-01

    Background Increasing the living-donor pool by accepting donors with an isolated medical abnormality (IMA) can significantly decrease the huge gap between limited supply and rising demand for organs. There is a wide range of variation among different centres in dealing with these categories of donors. We reviewed studies discussing living kidney donors with IMA, including greater age, obesity, hypertension, microscopic haematuria and nephrolithiasis, to highlight the effect of these abnormalities on both donor and recipient sides from medical and surgical perspectives. Methods We systematically searched MEDLINE, ISI Science Citation Index expanded, and Google scholar, from the inception of each source to January 2011, using the terms 慿idney transplant, 憆enal, 慻raft, 憀iving donor, 憃ld, 憃besity, 憂ephrolithiasis, 慼aematuria and 慼ypertension. In all, 58 studies were found to be relevant and were reviewed comprehensively. Results Most of the reviewed studies confirmed the safety of using elderly, moderately obese and well-controlled hypertensive donors. Also, under specific circumstances, donors with nephrolithiasis can be accepted. However, persistent microscopic haematuria should be considered seriously and renal biopsy is indicated to exclude underlying renal disease. Conclusion Extensive examination and cautious selection with tailored immunosuppressive protocols for these groups can provide a satisfactory short- and medium-term outcome. Highly motivated elderly, obese, controlled hypertensive and the donor with a unilateral small stone (<1.5燾m, with normal metabolic evaluation) could be accepted. Donors with dysmorphic and persistent haematuria should not be accepted. A close follow-up after donation is crucial, especially for obese donors who developed microalbuminuria.

  16. Microsphere-Based Immunoassay for the Detection of Azaspiracids

    PubMed Central

    Rodr韌uez, Laura P.; Vilari駉, Natalia; Louzao, M. Carmen; Dickerson, Tobin J.; Nicolaou, K. C.; Frederick, Michael O.; Botana, Luis M.

    2014-01-01

    Azaspiracids (AZAs) are a group of lipophilic toxins discovered in mussels from Ireland in 1995 following a human poisoning incident. Nowadays the regulatory limit for AZAs in many countries is set at 160 Fg of azaspiracid equivalents per kg of shellfish meat. In this work a microsphere-based immunoassay has been developed for the detection of AZAs using a Luminex system. This method is based on the competition between AZA-2 immobilized onto the surface of microspheres and free AZAs for the interaction with a monoclonal anti-azaspiracid antibody (mAb 8F4). In this inhibition immunoassay the amount of mAb 8F4 bound to AZA-2-microspheres was quantified using a phycoerythrin-labeled anti-mouse antibody, and the fluorescence was measured with a Luminex analyzer. Simple acetate/methanol or methanol extractions yielded final extracts with no matrix interferences and adequate recovery rates of 86.5% and 75.8%, respectively. In summary, this work presents, a sensitive and easily performed screening method capable of detecting AZAs at concentrations below the range of the European regulatory limit using a microsphere/flow cytometry system. PMID:24215909

  17. Finger-Actuated, Self-Contained Immunoassay Cassettes

    PubMed Central

    Qiu, Xianbo; Thompson, Jason A.; Chen, Zongyuan; Liu, Changchun; Chen, Dafeng; Ramprasad, Sudhir; Mauk, Michael G.; Ongagna, Serge; Barber, Cheryl; Abrams, William R.; Malamud, Daniel; Corstjens, Paul L.A.M.; Bau, Haim H.

    2010-01-01

    The building blocks for an inexpensive, disposable, luminescence-based microfluidic immunoassay cassette are described, and their integration in a point-of-care diagnostic system is demonstrated. Fluid motion in the cassette is driven by depressing finger-actuated pouches. All reagents needed for the immunoassay can be stored in the cassette in liquid form. Prior to use, the cassette consists of two separate parts. A top storage component contains pouches, sealed storage chambers, a metering chamber, and needle seats. The bottom processing component contains connection needles, a mixing chamber, and a detection chamber with immobilized proteins. Subsequent to sample introduction, the storage and processing components are mated. The needles form hydraulic connections between the two parts and, in some cases, close valves. The pouches are then actuated sequentially to induce flow of various reagents and facilitate process operations. The cassette is compatible with different detection modalities. Both a cassette with immunochromatographic-based detection and a cassette with microbead-based detection were constructed and evaluated. The immunochromatographic cassette was used to detect antibodies to HIV in saliva samples. The bead-based cassette was used to detect the proinflammatory chemokine IL-8. The experimental data demonstrates good repeatability and reasonable sensitivity. PMID:19597994

  18. Enzyme immunoassay validation for qualitative detection of cocaine in sweat.

    PubMed

    Spiehler, V; Fay, J; Fogerson, R; Schoendorfer, D; Niedbala, R S

    1996-01-01

    A solid-phase enzyme immunoassay (EIA) involving microtiter plates was modified for analysis of cocaine in sweat. Sweat was collected with the PharmChek sweat patch and drugs were eluted from the collection pad of the patch. The sweat contained primarily parent cocaine. The assay was determined to have cross-reactivity for cocaine of 102% relative to 100% for the benzoylecgonine (BE) calibrators and for cocaethylene of 148%. The optimum cutoff concentration for this modified assay, determined by receiver-operating characteristic curve analysis, was 10 micrograms/L cocaine or BE equivalents. At this concentration the assay had 94.5% sensitivity and 99.1% specificity vs gas chromatography-mass spectrometry (GC-MS) as an acceptable indicator of the true clinical state. The positive predictive value at a prevalence of 50% was 99%. Threshold analysis for positives suggested that the 95% confidence interval for a positive result by the EIA was between 12.5 and 15 micrograms/L and that quality-control samples at 5 and 15 micrograms/L could be run with each batch to certify the precision around the cutoff. All positive samples must be confirmed by GC-MS. The sensitivity and specificity of the overall analysis system (immunoassay screen and GC-MS confirmation) was 86% and 97%, with known cocaine dosing of volunteers as the acceptable indicator of the true clinical state. PMID:8565229

  19. Multiplex detection of plant pathogens using a microsphere immunoassay technology.

    PubMed

    Charlermroj, Ratthaphol; Himananto, Orawan; Seepiban, Channarong; Kumpoosiri, Mallika; Warin, Nuchnard; Oplatowska, Michalina; Gajanandana, Oraprapai; Grant, Irene R; Karoonuthaisiri, Nitsara; Elliott, Christopher T

    2013-01-01

    Plant pathogens are a serious problem for seed export, plant disease control and plant quarantine. Rapid and accurate screening tests are urgently required to protect and prevent plant diseases spreading worldwide. A novel multiplex detection method was developed based on microsphere immunoassays to simultaneously detect four important plant pathogens: a fruit blotch bacterium Acidovorax avenae subsp. citrulli (Aac), chilli vein-banding mottle virus (CVbMV, potyvirus), watermelon silver mottle virus (WSMoV, tospovirus serogroup IV) and melon yellow spot virus (MYSV, tospovirus). An antibody for each plant pathogen was linked on a fluorescence-coded magnetic microsphere set which was used to capture corresponding pathogen. The presence of pathogens was detected by R-phycoerythrin (RPE)-labeled antibodies specific to the pathogens. The assay conditions were optimized by identifying appropriate antibody pairs, blocking buffer, concentration of RPE-labeled antibodies and assay time. Once conditions were optimized, the assay was able to detect all four plant pathogens precisely and accurately with substantially higher sensitivity than enzyme-linked immunosorbent assay (ELISA) when spiked in buffer and in healthy watermelon leaf extract. The assay time of the microsphere immunoassay (1 hour) was much shorter than that of ELISA (4 hours). This system was also shown to be capable of detecting the pathogens in naturally infected plant samples and is a major advancement in plant pathogen detection. PMID:23638044

  1. Sensitive Immunoassays of Nitrated Fibrinogen in Human Biofluids

    SciTech Connect

    Tang, Zhiwen; Wu, Hong; Du, Dan; Wang, Jun; Wang, Hua; Qian, Weijun; Bigelow, Diana J.; Pounds, Joel G.; Smith, Richard D.; Lin, Yuehe

    2010-05-05

    Three new sandwich immunoassays for detection of nitrated biomarker have been established with potential applications in biomedical studies and clinical practice. In this study, nitrated human fibrinogen, a potential oxidative stress biomarker for several pathologies, was chosen as the target. To improve the sensitivity and overcome the interference caused by the complexity of human biofluids, we developed three sandwich strategies using various combinations of primary antibody and secondary antibody. All three strategies demonstrated high sensitivity and selectivity towards nitrated forms of fibrinogen in buffer, but their performances were dramatically reduced when tested with human plasma and serum samples. Systematically optimizations were carried out to investigate the effects of numerous factors, including sampling, coating, blocking, and immunoreactions. Our final optimization results indicate that two of these strategies retain sufficient sensitivity and selectivity for use as assays in human physiological samples. Specifically, detection limits reached the pM level and the linear response ranges were up to nM level with a correlation coefficient > 0.99. To our best knowledge, this is the first example of using an electrochemical immunoassay for a nitrated biomarker in a physiological fluid. This novel approach provides a rapid, sensitive, selective, cost efficient and robust bioassay for detection of oxidative stress in pathology and for clinical applications. Moreover, the sandwich strategies developed in this paper can be readily used to establish effective methods targeting other nitration biomarkers.

  2. Aequorin fusion proteins as bioluminescent tracers for competitive immunoassays

    NASA Astrophysics Data System (ADS)

    Mirasoli, Mara; Michelini, Elisa; Deo, Sapna K.; Dikici, Emre; Roda, Aldo; Daunert, Sylvia

    2004-06-01

    The use of bio- and chemiluminescence for the development of quantitative binding assays offers undoubted advantages over other detection systems, such as spectrophotometry, fluorescence, or radioactivity. Indeed, bio- and chemiluminescence detection provides similar, or even better, sensitivity and detectability than radioisotopes, while avoiding the problems of health hazards, waste disposal, and instability associated with the use of radioisotopes. Among bioluminescent labels, the calcium-activated photoprotein aequorin, originally isolated from Aequorea victoria and today available as a recombinant product, is characterized by very high detectability, down to attomole levels. It has been used as a bioluminescent label for developing a variety of highly sensitive immunoassays, using various analyte-aequorin conjugation strategies. When the analyte is a protein or a peptide, genetic engineering techniques can be used to produce protein fusions where the analyte is in-frame fused with aequorin, thus producing homogeneous one-to-one conjugation products, available in virtually unlimited amount. Various assays were developed using this strategy: a short review of the most interesting applications is presented, as well as the cloning, purification and initial characterization of an endothelin-1-aequorin conjugate suitable for developing a competitive immunoassay for endothelin-1, a potent vasoconstrictor peptide, involved in hypertension.

  3. Quantum-Dots Based Electrochemical Immunoassay of Interleukin-1?

    SciTech Connect

    Wu, Hong; Liu, Guodong; Wang, Jun; Lin, Yuehe

    2007-07-01

    We describe a quantum-dot (QD, CdSe@ZnS)-based electrochemical immunoassay to detect a protein biomarker, interleukin-1? (IL-1?). QD conjugated with anti-IL-1? antibody was used as a label in an immunorecognition event. After a complete sandwich immunoreaction among the primary IL-1? antibody (immobilized on the avidin-modified magnetic beads), IL-1?, and the QD-labeled secondary antibody, QD labels were attached to the magnetic-bead surface through the antibody-antigen immunocomplex. Electrochemical stripping analysis of the captured QDs was used to quantify the concentration of IL-1? after an acid-dissolution step. The streptavidin-modified magnetic beads and the magnetic separation platform were used to integrate a facile antibody immobilization (through a biotin/streptavidin interaction) with immunoreactions and the isolation of immunocomplexes from reaction solutions in the assay. The voltammetric response is highly linear over the range of 0.5 to 50 ng mL-1 IL 1?, and the limit of detection is estimated to be 0.3 ng mL-1 (18 pM). This QD-based electrochemical immunoassay shows great promise for rapid, simple, and cost-effective analysis of protein biomarkers.

  4. Nanobody medicated immunoassay for ultrasensitive detection of cancer biomarker alpha-fetoprotein.

    PubMed

    Chen, Jing; He, Qing-Hua; Xu, Yang; Fu, Jin-Heng; Li, Yan-Ping; Tu, Zhui; Wang, Dan; Shu, Mei; Qiu, Yu-Lou; Yang, Hong-Wei; Liu, Yuan-Yuan

    2016-01-15

    Immunoassay for cancer biomarkers plays an important role in cancer prevention and early diagnosis. To the development of immunoassay, the quality and stability of applied antibody is one of the key points to obtain reliability and high sensitivity for immunoassay. The main purpose of this study was to develop a novel immunoassay for ultrasensitive detection of cancer biomarker alpha-fetoprotein (AFP) based on nanobody against AFP. Two nanobodies which bind to AFP were selected from a phage display nanobody library by biopanning strategy. The prepared nanobodies are clonable, thermally stable and applied in both sandwich enzyme linked immunoassay (ELISA) and immuno-PCR assay for ultrasensitive detection of AFP. The limit detection of sandwich ELISA setup with optimized nanobodies was 0.48ngmL(-1), and the half of saturation concentration (SC50) value was 6.680.56ngmL(-1). These nanobodies were also used to develop an immuno-PCR assay for ultrasensitive detection of AFP, its limit detection values was 0.005ngmL(-1), and the linear range was 0.01-10,000ngmL(-1). These established immunoassays based on nanobodies were highly specific to AFP and with negligible cross reactivity with other tested caner biomarkers. Furthermore, this novel concept of nanobodies mediated immunoassay may provide potential applications in a general method for the ultrasensitive detection of various cancer biomarkers. PMID:26592642

  5. Kinetic analyses and performance of a colloidal magnetic nanoparticle based immunoassay dedicated to allergy diagnosis.

    PubMed

    Teste, Bruno; Kanoufi, Fr閐閞ic; Descroix, St閜hanie; Poncet, Pascal; Georgelin, Thomas; Siaugue, Jean-Michel; Petr, Jan; Varenne, Anne; Hennion, Marie-Claire

    2011-07-01

    In this paper, we demonstrate the possibility to use magnetic nanoparticles as immunosupports for allergy diagnosis. Most immunoassays used for immunosupports and clinical diagnosis are based on a heterogeneous solid-phase system and suffer from mass-transfer limitation. The nanoparticles' colloidal behavior and magnetic properties bring the advantages of homogeneous immunoassay, i.e., species diffusion, and of heterogeneous immunoassay, i.e., easy separation of the immunocomplex and free forms, as well as analyte preconcentration. We thus developed a colloidal, non-competitive, indirect immunoassay using magnetic core-shell nanoparticles (MCSNP) as immunosupports. The feasibility of such an immunoassay was first demonstrated with a model antibody and described by comparing the immunocapture kinetics using macro (standard microtiter plate), micro (microparticles) and nanosupports (MCSNP). The influence of the nanosupport properties (surface chemistry, antigen density) and of the medium (ionic strength, counter ion nature) on the immunocapture efficiency and specificity was then investigated. The performances of this original MCSNP-based immunoassay were compared with a gold standard enzyme-linked immunosorbent assay (ELISA) using a microtiter plate. The capture rate of target IgG was accelerated 200-fold and a tenfold lower limit of detection was achieved. Finally, the MCSNP-based immunoassay was successfully applied to the detection of specific IgE from milk-allergic patient's sera with a lower LOD and a good agreement (CV?

  6. Seroepidemiology of Toxoplasma gondii Infection among Healthy Blood Donors in Taiwan

    PubMed Central

    Chiang, Ting-Yi; Hsieh, Hwei-Ho; Kuo, Ming-Chu; Chiu, Kai-Tse; Lin, Wei-Chen; Fan, Chia-Kwung; Fang, Chi-Tai; Ji, Dar-Der

    2012-01-01

    Toxoplasma gondii is an opportunistic, zoonotic pathogen with a worldwide distribution. There are large variations in the seroprevalence of T. gondii infection in different regions of the world. Although toxoplasmosis became a notifiable communicable disease in Taiwan in 2007, little is known about its epidemiology among the general population. This cross-sectional study aimed to survey the seroprevalence of T. gondii infection and its risk factors among healthy blood donors in Taiwan. Through collaborating with the Taiwan Blood Services Foundation, a total of 1,783 healthy blood donors from all six-branch blood service centers participated in this study. The blood samples were tested for the presence of T. gondii antibodies and DNA using enzyme immunoassays and real-time PCR, respectively. Structured questionnaires were used to gather information on risk factors for T. gondii infection. Of the 1,783 participants, 166 (9.3%) tested positive for anti-Toxoplasma IgG, while 5 (0.28%) tested positive for anti-Toxoplasma IgM. The five IgM positive donors had high avidity antibodies suggestive of past infection. No active parasitemia was detected by real-time PCR assays. Multivariate logistic regression showed that undercooked pork meat consumption (adjusted odds ratio [OR]?=?2.9; 95% confidence interval [CI]: 1.36.5), raw mussels consumption (adjusted OR?=?5.3; 95% CI: 1.519.1), having a cat in the household (adjusted OR?=?2.0; 95% CI: 1.23.2), a lower education level (adjusted OR?=?1.6; 95% CI: 1.12.3), and donation place in eastern Taiwan (adjusted OR?=?2.5; 95% CI: 1.63.9) were independent risk factors for Toxoplasma seropositivity. These findings provide information on the seroprevalence and epidemiology of T. gondii infection among healthy blood donors in Taiwan. PMID:23133557

  7. Gamete donors' expectations and experiences of contact with their donor offspring

    PubMed Central

    Kirkman, Maggie; Bourne, Kate; Fisher, Jane; Johnson, Louise; Hammarberg, Karin

    2014-01-01

    STUDY QUESTION What are the expectations and experiences of anonymous gamete donors about contact with their donor offspring? SUMMARY ANSWER Rather than consistently wanting to remain distant from their donor offspring, donors' expectations and experiences of contact with donor offspring ranged from none to a close personal relationship. WHAT IS KNOWN ALREADY Donor conception is part of assisted reproduction in many countries, but little is known about its continuing influence on gamete donors' lives. STUDY DESIGN, SIZE, DURATION A qualitative research model appropriate for understanding participants' views was employed; semi-structured interviews were conducted during January朚arch 2013. PARTICIPANTS/MATERIALS, SETTING, METHODS Before 1998, gamete donors in Victoria, Australia, were subject to evolving legislation that allowed them to remain anonymous or (from 1988) to consent to the release of identifying information. An opportunity to increase knowledge of donors' expectations and experiences of contact with their donor offspring recently arose in Victoria when a recommendation was made to introduce mandatory identification of donors on request from their donor offspring, with retrospective effect. Pre-1998 donors were invited through an advertising campaign to be interviewed about their views, experiences and expectations; 36 sperm donors and 6 egg donors participated. MAIN RESULTS AND THE ROLE OF CHANCE This research is unusual in achieving participation by donors who would not normally identify themselves to researchers or government inquiries. Qualitative thematic analysis revealed that most donors did not characterize themselves as parents of their donor offspring. Donors' expectations and experiences of contact with donor offspring ranged from none to a close personal relationship. LIMITATIONS, REASONS FOR CAUTION It is not possible to establish whether participants were representative of all pre-1998 donors. WIDER IMPLICATIONS OF THE FINDINGS Anonymous donors' needs and desires are not homogeneous; policy and practice should be sensitive and responsive to a wide range of circumstances and preferences. Decisions made to restrict or facilitate contact or the exchange of information have ramifications for donors as well as for donor-conceived people. STUDY FUNDING/COMPETING INTEREST(S) The study was funded by the Victorian Department of Health. The authors have no conflicts of interest to declare. TRIAL REGISTRATION NUMBER Not applicable. PMID:24549216

  8. Donor free radical explosive composition

    DOEpatents

    Walker, Franklin E. [15 Way Points Rd., Danville, CA 94526; Wasley, Richard J. [4290 Colgate Way, Livermore, CA 94550

    1980-04-01

    An improved explosive composition is disclosed and comprises a major portion of an explosive having a detonation velocity between about 1500 and 10,000 meters per second and a minor amount of a donor additive comprising an organic compound or mixture of organic compounds capable of releasing low molecular weight free radicals or ions under mechanical or electrical shock conditions and which is not an explosive, or an inorganic compound or mixture of inorganic compounds capable of releasing low molecular weight free radicals or ions under mechanical or electrical shock conditions and selected from ammonium or alkali metal persulfates.

  9. Exxon donor solvent liquefaction process

    NASA Astrophysics Data System (ADS)

    Neavel, R. C.

    1981-03-01

    The Exxon donor solvent (EDS) coal liquefaction system is a direct liquefaction procedure. Coal is chemically reacted and dissolved in a recycle solvent that is hydrogenated between passes to the liquefaction reactor. More than 2.6 barrels of a synthetic crude boiling below 1000 F are produced per ton of dry, high volatile coal feed. Other ranks of coal can be effectively liquefied. The process development has proceeded to a 250 ton/day pilot plant stage that went into operation in June 1980. The presentation addresses the chemical reactions and process conditions that result in ease of operability and flexibility of the EDS process.

  10. Development of a Disperse Dye Immunoassay Technique for Detection of Antibodies against Neospora caninum in Cattle

    PubMed Central

    Selahi, Fatemeh; Hosseini, Mohammad Hossein; Mansourian, Maryam; Tahamtan, Yahya

    2013-01-01

    In this study a disperse dye immunoassay method was standardized and evaluated for detection of antibodies against Neospora caninum in cattle. Sera from 150 cattle with a recent history of abortion were collected and tested by commercial ELISA kit and a standardized in-house dye immunoassay system. The positivity rate for the sera used in this study was 34.6% for the disperse dye immunoassay (DDIA) compared to 32% obtained by ELISA kit. This study showed no significant difference between DDIA and ELISA. The results indicated that the DDIA provide an economic, simple, rapid and robust test for detection of N. caninum infection in cattle. PMID:23467930

  11. Multiplexed immunoassays for biomonitoring of exposure to agrochemicals using quantum dots as fluorescent reporters

    NASA Astrophysics Data System (ADS)

    Nichkova, Mikaela; Dosev, Dosi; Davies, Alexander E.; Gee, Shirley J.; Kennedy, Ian M.; Hammock, Bruce D.

    2007-02-01

    The application of quantum dots (QDs) as labels in immunoassay microarrays for the multiplex detection of 3- phenoxybenzoic acid (PBA) and atrazine-mercapturate (AM) has been demonstrated. PBA and AM are biomarkers of exposure to the pyrethroid insecticides and to the herbicide atrazine, respectively. Microarrays were fabricated by microcontact printing of the coating antigens in line patterns onto glass substrates. Competitive immunoassays were successfully performed using quantum dots (QD560 and QD620) as reporters. The multiplexed immunoassays were characterized by fluorescence microscopy and SEM. The application of QD fluorophores facilitates multiplex assays and therefore can contribute to enhanced throughput in biomonitoring.

  12. Long-term outcomes of kidney donors

    PubMed Central

    Morgan, Benjamin R.; Ibrahim, Hassan N.

    2011-01-01

    As the demand for kidney transplantation, particularly from living donors, continues to rise, there is increasing and much needed interest in accurately quantifying the long-term risks of kidney donation. We review the outcomes of kidney donors in the domains of survival, perioperative mortality, risk of end-stage renal disease, quality of life, course of diabetes mellitus in donors, pregnancy after donation, obesity, and prevalence of other health conditions.

  13. [Kinds and characteristics of solid-phase enzyme immunoassay].

    PubMed

    Sugiura, M; Mizuoka, K

    1995-09-01

    Technology with solid-phase enzyme immunoassay was markedly progressed. Kinds of solid-phase are as follows; (1) tube, (2) beads, (3) latex particle, (4) microtiter plate, (5) ferrite particle, (6) membrane. Recently, application of avidin/biotin system to the tube method was accomplished. As one of the B/F separation with beads method, spin washing method was also developed. Automated open system with the beads method was also developed. Microtiter plate using sponges as the solid-phase was routinely used for allergy tests (CAP-RAST). Multi-Access Open System (MAOS), applicable for various kinds of ELISA kit, will appear in Japan near future. Latex particle with ferrite plating and ferrite particle with glass coating were also used as the solid-phase of EIA. HCV-RIBA II and III systems using nitro-cellulose membrane as solid-phase were also described. PMID:7474379

  14. Chemiluminescence enzyme immunoassay for the determination of sulfamethoxydiazine

    NASA Astrophysics Data System (ADS)

    Wu, Yongjun; Yu, Songcheng; Yu, Fei; Yan, Nali; Qu, Lingbo; Zhang, Hongquan

    2011-10-01

    Sulfamethoxydiazine (SMD), which is often used for animal disease treatment, is harmful to human health. No SMD residue should be detected in food in some countries, such as USA and Japan. Therefore, it is significant to develop a high-throughput, high-sensitivity and accurate method for the determination of the content of SMD in food. In this paper, chemiluminescence enzyme immunoassay (CLEIA) was developed for quantification of SMD. For this method, the limit of detection was 3.2 pg/ml, the linear range was from 10 to 2000 pg/ml, the within-day and inter-day precision were below 13% and below 18%, respectively, and the recovery was from 85% to 105%. Milk and egg were selected as samples to be examined with this method, and the result indicated that this CLEIA method was suitable for screening and quality control of food.

  15. Development of an Ultrasensitive Immunoassay for Detecting Tartrazine

    PubMed Central

    Li, Zhuokun; Song, Shanshan; Xu, Liguang; Kuang, Hua; Guo, Shidong; Xu, Chuanlai

    2013-01-01

    We have developed an ultrasensitive indirect competitive enzyme-linked immunosorbent assay for the determination of tartrazine. Two carboxylated analogues of tartrazine with different spacer lengths, and one derivative from commercial tartrazine after a little chemical modification, were synthesized as haptens in order to produce antibodies specific to tartrazine. The effect of sulfonic acid groups on the hapten structure of tartrazine was also studied carefully for the first time. A most specific monoclonal antibody against tartrazine was created and exhibited an IC50 value of 0.105 ng/mL and a limit of detection of 0.014 ng/mL, with no cross-reactivity to other structurally-related pigments. The established immunoassay was applied to the determination of tartrazine in fortified samples of orange juice and in real positive samples of carbonated beverages. PMID:23799494

  16. IgE Epitope Mapping Using Peptide Microarray Immunoassay.

    PubMed

    Gimenez, Gustavo; Bened, Sara; Lin, Jing

    2016-01-01

    IgE epitope mapping of food allergens contributes to a better understanding of the pathogenesis of food allergy and may become an additional tool for food allergy diagnosis/prognosis. Microarray platforms which allow for simultaneous screening of a large number of peptides corresponding to the sequences of food allergens are ideally suited for large-scale IgE epitope mapping. Here we describe the method of performing a reliable and sensitive peptide microarray immunoassay, which was developed in our lab and results in the identification of candidate IgE epitope biomarkers useful in determining allergic disease severity and prognosis, as well as in the prediction of treatment outcomes. A gastric digestion model that can be applied to prescreen peptides and reduce costs in the peptide microarray is also described in this chapter. PMID:26490481

  17. Varying results for immunoassay screening kits for cotinine level.

    PubMed

    Alterman, Arthur I; Gariti, Peter; Niedbala, R Sam

    2002-09-01

    Our earlier study found that although enzyme-linked immunosorbent analysis (ELISA) screening assays for urine cotinine indicated use in former smoking treatment patients who reported abstinence, this finding was sometimes incorrect when validated against gas chromatography/mass spectrometry (GC/ MS; P. Gariti, A. I. Alterman, R. Ehrmann, F. D. Mulvaney, & C. P. O'Brien, 2002). In the current validation study, separate urine samples of 71 of these same patients were reanalyzed by an independent laboratory in blinded fashion using a screening enzyme immunoassay (EIA) analysis and GC/MS confirmation. EIA results showed almost total agreement with confirmatory testing. The findings indicate that use of screening ELISA/EIA for urine cotinine can detect unreported cases of smoking in former patients, but that care is needed in selecting a laboratory for conducting these tests. PMID:12236461

  18. Biosensor immunoassay for traces of hazelnut protein in olive oil

    PubMed Central

    Smits, Nathalie G. E.; Haasnoot, Willem

    2009-01-01

    The fraudulent addition of hazelnut oil to more expensive olive oil not only causes economical loss but may also result in problems for allergic individuals as they may inadvertently be exposed to potentially allergenic hazelnut proteins. To improve consumer safety, a rapid and sensitive direct biosensor immunoassay, based on a highly specific monoclonal antibody, was developed to detect the presence of hazelnut proteins in olive oils. The sample preparation was easy (extraction with buffer); the assay time was fast (4.5爉in only) and the limit of detection was low (0.08?g/g of hazelnut proteins in olive oil). Recoveries obtained with an olive oil mixed with different amounts of a hazelnut protein containing hazelnut oil varied between 93% and 109%. Electronic supplementary material The online version of this article (doi:10.1007/s00216-009-2720-1) contains supplementary material, which is available to authorized users. PMID:19263041

  19. Performance of immunoassay kits for site characterization and remediation

    SciTech Connect

    Waters, L.C.; Palausky, A.; Counts, R.W.; Jenkins, R.A.

    1995-12-31

    The US Department of Energy (DOE) is supporting efforts to identify, validate and implement the use of effective, low-cost alternatives to currently used analytical methods for environmental management. As part of that program, we have evaluated the performances of a number of immunoassay (IA) kits with specificities for environmental contaminants of concern to the DOE. The studies were done in the laboratory using both spiked and field test samples. The analyte specificity and manufacturers of the kits evaluated were the following: mercury, BioNebraska; polychlorinated biphenyls (PCBs), EnSys and Millipore; petroleum fuel hydrocarbons, Millipore and Ohmicron; and polyaromatic hydrocarbons (PAHs), Ohmicron and Millipore. The kits were used in either a semiquantitative or quantitative format according to the preference of the manufacturers.

  20. Alternative Donor Transplantation for Acute Myeloid Leukemia

    PubMed Central

    Bejanyan, Nelli; Haddad, Housam; Brunstein, Claudio

    2015-01-01

    Allogeneic hematopoietic cell transplantation (allo-HCT) is a potentially curative therapy for adult patients with acute myeloid leukemia (AML), but its use for consolidation therapy after first remission with induction chemotherapy used to be limited to younger patients and those with suitable donors. The median age of AML diagnosis is in the late 60s. With the introduction of reduced-intensity conditioning (RIC), many older adults are now eligible to receive allo-HCT, including those who are medically less fit to receive myeloablative conditioning. Furthermore, AML patients commonly have no human leukocyte antigen (HLA)-identical or medically suitable sibling donor available to proceed with allo-HCT. Technical advances in donor matching, suppression of alloreactivity, and supportive care have made it possible to use alternative donors, such as unrelated umbilical cord blood (UCB) and partially HLA-matched related (haploidentical) donors. Outcomes after alternative donor allo-HCT are now approaching the outcomes observed for conventional allo-HCT with matched related and unrelated donors. Thus, with both UCB and haploidentical donors available, lack of donor should rarely be a limiting factor in offering an allo-HCT to adults with AML. PMID:26239557

  1. Hybrid super electron donors preparation and reactivity

    PubMed Central

    Garnier, Jean; Thomson, Douglas W; Zhou, Shengze; Jolly, Phillip I; Berlouis, Leonard E A

    2012-01-01

    Summary Neutral organic electron donors, featuring pyridinylidene杋midazolylidene, pyridinylidene朾enzimidazolylidene and imidazolylidene朾enzimidazolylidene linkages are reported. The pyridinylidene朾enzimidazolylidene and imidazolylidene朾enzimidazolylidene hybrid systems were designed to be the first super electron donors to convert iodoarenes to aryl radicals at room temperature, and indeed both show evidence for significant aryl radical formation at room temperature. The stronger pyridinylidene杋midazolylidene donor converts iodoarenes to aryl anions efficiently under appropriate conditions (3 equiv of donor). The presence of excess sodium hydride base has a very important and selective effect on some of these electron-transfer reactions, and a rationale for this is proposed. PMID:23019427

  2. Capillary electrophoretic enzyme immunoassay for digoxin in human serum.

    PubMed

    Liu, X; Xu, Y; Ip, M P

    1995-09-15

    The combined use of capillary electrophoretic (CE) separation and homogeneous enzyme immunoassay for analyzing drugs in hemolyzed, lipemic, or icteric serum samples was investigated. An FDA-approved EMIT assay kit for digoxin in human serum was used. After the enzyme immunoassay, the enzymatic reaction product (NADH) and remaining substrate (NAD+), together with internal standard (p-nitrophenol, NP), were electrokinetically injected into a polyacrylamide-coated electrophoresis capillary and separated under applied potential. Detection was made by monitoring the UV adsorption at wavelength of 260 nm. The digoxin level in human serum was determined by comparing the peak area ratio of NADH and NP to the ratios established by the known digoxin standards. In this study, the factors that influence the CE separation were also investigated. Under the optimum conditions, NADH, NAD+, and NP were separated at electric field strength of 438 V/cm in the coated capillary (100 microns x 57 cm) with 200 mM Tris-borate buffer (pH 7.9) containing 0.2% hydroxypropyl methylcellulose. CE analyses of serum samples spiked with NADH standards at concentrations of 100 and 400 microM resulted in detection variabilities of less than 2% and analytical recoveries of 98-102%. Both an internal calibration plot for NADH and a dose-response curve for digoxin in serum were constructed. Calibrator serum, patients' sera with hemolyzed, lipemic, and icteric interference factors, and other pigmented blood components (e.g., serum albumin, bilirubin, hemoglobin, uric acid, coproporphyrin, melanin, protoporphyrin IX, and uroprophyrin) demonstrated no interference in this method. The authors believed that this method is useful for analyzing digoxin in hemolyzed, lipemic, and icteric blood samples that are known to create problems in conventional EMIT assays and may be applicable to other EMIT-based assays for monitoring drugs in complex biological matrices. PMID:8686886

  3. Automated homogeneous immunoassay analysis of cotinine in urine.

    PubMed

    Niedbala, R Sam; Haley, Nancy; Kardos, Stephanie; Kardos, Keith

    2002-04-01

    A study was conducted to evaluate the performance comparison of a homogeneous enzyme immunoassay (EIA) designed to detect cotinine in urine and carbon monoxide (CO) breath measurements to determine smoking status. The clinical comparison was done using urine and breath specimens from 218 volunteers. Urine samples were analyzed by immunoassay and confirmed by gas chromatography-mass spectrometry (GC-MS). Breath carbon monoxide was determined by a commercial analyzer. Using cutoffs of 10 ppm for CO and 500 ng/mL for urinary cotinine, the relative sensitivity/specificity was 93.6%/74.0%. The positive predictive value was 86.8%, and the negative predictive value was 86.5%. However, comparison of the EIA to GC-MS showed a sensitivity/specificity of 96.2%/98.4% and a positive predictive value of 99.3%. The EIA was also evaluated non-clinically for precision, stability, recovery, and interferences. In addition, the non-clinical evaluation demonstrated coefficients of variation from 0.37 to 1.09% across cotinine concentrations ranging from 0 to 5000 ng/mL. The assay was found to be highly specific for cotinine and cross-reacted to a limited degree with 3-hydroxycotinine. Finally, multiple freeze-thaw cycles of urines containing cotinine showed no degradation of the drug in the specimen when tested in the EIA. Thus, the EIA tested is a rapid, lab-based test that can reliably determine cotinine levels and their relation to smoking status. PMID:11991533

  4. Validation of an automated enzyme immunoassay for the measurement of serum total thyroxine in cats

    E-print Network

    Williams, Tim L.; Archer, Joy

    2015-05-18

    immunoassay (EIA) for use with feline serum and derivation of a TT4 reference interval (RI) for cats aged 9 years and older). Methods: Assay precision, reproducibility, and linearity were evaluated. Interference by hemolysis was also assessed. Method...

  5. An Inexpensive, Fast and Sensitive Quantitative Lateral Flow Magneto-Immunoassay for Total Prostate Specific Antigen

    PubMed Central

    Barnett, Jacqueline M.; Wraith, Patrick; Kiely, Janice; Persad, Raj; Hurley, Katrina; Hawkins, Peter; Luxton, Richard

    2014-01-01

    We describe the detection characteristics of a device the Resonant Coil Magnetometer (RCM) to quantify paramagnetic particles (PMPs) in immunochromatographic (lateral flow) assays. Lateral flow assays were developed using PMPs for the measurement of total prostate specific antigen (PSA) in serum samples. A detection limit of 0.8 ng/mL was achieved for total PSA using the RCM and is at clinically significant concentrations. Comparison of data obtained in a pilot study from the analysis of serum samples with commercially available immunoassays shows good agreement. The development of a quantitative magneto-immunoassay in lateral flow format for total PSA suggests the potential of the RCM to operate with many immunoassay formats. The RCM has the potential to be modified to quantify multiple analytes in this format. This research shows promise for the development of an inexpensive device capable of quantifying multiple analytes at the point-of-care using a magneto-immunoassay in lateral flow format. PMID:25587419

  6. Multiplexed immunoassays for proteins using magnetic luminescent nanoparticles for internal calibration

    E-print Network

    Hammock, Bruce D.

    Multiplexed immunoassays for proteins using magnetic luminescent nanoparticles for internalO3/luminescent Eu:Gd2O3 core/ shell nanoparticles (MLNPs) as a carrier. The magnetic properties. All rights reserved. Keywords: Luminescence; Nanoparticles; Lanthanide oxide; Magnetic; Multiplex

  7. Correction: Ultrafast immunoassays by coupling dielectrophoretic biomarker enrichment in nanoslit channel with electrochemical detection on graphene.

    PubMed

    Sanghavi, Bankim J; Varhue, Walter; Rohani, Ali; Liao, Kuo-Tang; Bazydlo, Lindsay A L; Chou, Chia-Fu; Swami, Nathan S

    2015-12-21

    Correction for 'Ultrafast immunoassays by coupling dielectrophoretic biomarker enrichment in nanoslit channel with electrochemical detection on graphene' by Bankim J. Sanghavi et al., Lab Chip, 2015, DOI: . PMID:26528632

  8. Development of an integrated capillary valve-based preconcentrator and surface-based immunoassay

    E-print Network

    Liu, Vincent Hok

    2009-01-01

    A new generation of integrated preconcentrator and immunoassay was developed. A novel, self-aligned method for patterning Nafion resin was developed and applied to create a preconcentrator. In a parallel effort, a surface-based ...

  9. Immunoassay Techniques for Detection of the Herbicide Simazine Based on Use of Oppositely

    E-print Network

    Hammock, Bruce D.

    Immunoassay Techniques for Detection of the Herbicide Simazine Based on Use of Oppositely Charged to an assay for the herbicide simazine. Both enzyme-linked immu- nosorbent assay (ELISA) and dot blot formats

  10. Inadequate donor size in cadaver kidney transplantation.

    PubMed

    Kasiske, Bertram L; Snyder, Jon J; Gilbertson, David

    2002-08-01

    There have been conflicting reports that kidneys from small donors may be at increased risk for late graft failure if they are transplanted into large recipients. Data from the United States Renal Data System was used to study all first cadaver kidney transplantations performed during the years 1994 to 1999. Donor and recipient body surface area (BSA) combinations were included along with other patient and transplant characteristics in a Poisson analysis of factors associated with early (in the first 4 mo) and late (> or =4 mo) graft failure. The numbers of large (BSA >2.2 m(2)) and medium size (BSA 1.6 to 2.2 m(2)) recipients that received kidneys from small (BSA <1.6 m(2)) donors are less than expected (chi(2) = 118.09; P < 0.0001), suggesting that transplant centers may be refusing some kidneys on the basis of donor-recipient size differences. Large recipients who received kidneys from small donors made up 1.5% of the population and had a 43% (95% CI, 17 to 75%; P = 0.0004) increased risk of late graft failure compared with medium-size recipients who received kidneys from medium-size donors (53.4% of the population). Medium-size recipients who received kidneys from small donors made up 12.0% of the population and had a 16% (95% CI, 6 to 26%; P = 0.0012) increased risk of late graft failure. Disparities in recipient and donor size had similar adverse affects on mortality. Effects of recipient obesity (body mass index) and donor gender on late graft survival were no longer statistically significant after the effects of donor and recipient body size were taken into account. In conclusion, the relative size of the donor and recipient should possibly be taken into account when choosing kidneys for transplantation. PMID:12138149

  11. Horseradish Peroxidase-Mediated, Iodide-Catalyzed Cascade Reaction for Plasmonic Immunoassays.

    PubMed

    Xianyu, Yunlei; Chen, Yiping; Jiang, Xingyu

    2015-11-01

    This report outlines an enzymatic cascade reaction for signal transduction and amplification for plasmonic immunoassays by using horseradish peroxidase (HRP)-mediated aggregation of gold nanoparticles (AuNPs). HRP-catalyzed oxidation of iodide and iodide-catalyzed oxidation of cysteine is employed to modulate the plasmonic signals of AuNPs. It agrees well with the current immunoassay platforms and allows naked-eye readout with enhanced sensitivity, which holds great promise for applications in resource-constrained settings. PMID:26460152

  12. The Experience of Living Kidney Donors

    ERIC Educational Resources Information Center

    Brown, Judith Belle; Karley, Mary Lou; Boudville, Neil; Bullas, Ruth; Garg, Amit X.; Muirhead, Norman

    2008-01-01

    This article describes the experiences, feelings, and ideas of living kidney donors. Using a phenomenological, qualitative research approach, the authors interviewed 12 purposefully selected living kidney donors (eight men and four women), who were between four and 29 years since donation. Interviews were audiotaped, and transcribed verbatim, and

  13. Payment for donor kidneys: pros and cons.

    PubMed

    Friedman, E A; Friedman, A L

    2006-03-01

    Continuous growth of the end stage renal disease population treated by dialysis, outpaces deceased donor kidneys available, lengthens the waiting time for a deceased donor transplant. As estimated by the United States Department of Health & Human Services: '17 people die each day waiting for transplants that can't take place because of the shortage of donated organs.' Strategies to expand the donor pool--public relations campaigns and Drivers' license designation--have been mainly unsuccessful. Although illegal in most nations, and viewed as unethical by professional medical organizations, the voluntary sale of purchased donor kidneys now accounts for thousands of black market transplants. The case for legalizing kidney purchase hinges on the key premise that individuals are entitled to control of their body parts even to the point of inducing risk of life. One approach to expanding the pool of kidney donors is to legalize payment of a fair market price of about 40,000 dollars to donors. Establishing a federal agency to manage marketing and purchase of donor kidneys in collaboration with the United Network for Organ Sharing might be financially self-sustaining as reduction in costs of dialysis balances the expense of payment to donors. PMID:16482095

  14. Recipients' views on payment of sperm donors.

    PubMed

    Ravelingien, An; Provoost, Veerle; Wyverkens, Elia; Buysse, Ann; De Sutter, Petra; Pennings, Guido

    2015-08-01

    The aim of this qualitative study was to explore how recipients viewed payment of sperm donors. The study was conducted in Belgium, where, as in many countries, sperm donors receive recompense for their time and expenses. Face-to-face semi-structured interviews were conducted with 34 heterosexual and lesbian couples who, at the time of data collection, had at least one donor-conceived child aged 7-10 years or who were undergoing donor conception treatment. Although participants commonly described the issue of financial compensation as something that did not really concern them, all supported the idea that some level of payment was acceptable or even necessary. The participants also identified several ways in which donor payment offered advantages to their own position as (future) parents. Although the idea is commonly rehearsed that sperm donation is a gift and that monetary transaction for conception is demeaning, the participants of this study did not generally share this view. To them, a small financial return served as a symbolic acknowledgement of the donor's contribution and helped secure the type of relationship they expected from their donor. There was clearly concern, however, over high payments and the risk of attracting the wrong kind of donor. PMID:26099446

  15. Application of elderly donor for liver transplantation

    PubMed Central

    Li, Jiang; Wang, Kai

    2015-01-01

    Recently, much more attention has been paid on application of elderly donor due to the shortage of organs. Although liver quality of elderly donors may be sub-optimal comparing with that from younger donors, primary non-function of a liver graft is a rare event. On the other hand, long-term graft and recipient survival for usage of elderly grafts has become a major concern and focus of research. Many transplant centers have changed the upper limit of donor age from previous 50 to 70 or even 75-year-old and achieved good graft function. Although some scholars believed that liver transplant using elderly grafts was associated with high probability of delayed liver function recovery, graft loss and hepatitis C recurrence, reports from several transplant centers document that long-term survival of grafts and recipients may be significantly improved through certain screening of donors and recipients before transplant. In conclusion, it is very important and relatively safe to use grafts from elderly donors to expand the donor pool. However, elderly donors and corresponding recipients must be carefully selected before transplant. The long-term effect of advanced age on grafts and recipients need to be evaluated through a comprehensive and long-term in-depth observation. PMID:26379822

  16. 21 CFR 610.41 - Donor deferral.

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ... 21 Food and Drugs 7 2012-04-01 2012-04-01 false Donor deferral. 610.41 Section 610.41 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) BIOLOGICS GENERAL BIOLOGICAL PRODUCTS STANDARDS Testing Requirements for Communicable Disease Agents 610.41 Donor deferral. (a) You, an establishment that...

  17. [Diphtheria and tetanus immunity of blood donors].

    PubMed

    Zakrzewska, A

    1997-01-01

    Immunity to diphtheria and tetanus was determined in serum samples from 108 blood donors. Antitoxin concentration was evaluated by ELISA assay. 65% of donors were protected to diptheria and 100% to tetanus. Significant trend of decreasing immunity was observed with increasing age. PMID:9333856

  18. Kinetics of thermal donor generation in silicon

    NASA Technical Reports Server (NTRS)

    Mao, B.-Y.; Lagowski, J.; Gatos, H. C.

    1984-01-01

    The generation kinetics of thermal donors at 450 C in Czochralski-grown silicon was found to be altered by high-temperature preannealing (e.g., 1100 C for 30 min). Thus, when compared with as-grown Si, high-temperature preannealed material exhibits a smaller concentration of generated thermal donors and a faster thermal donor saturation. A unified mechanism of nucleation and oxygen diffusion-controlled growth (based on solid-state plate transformation theory) is proposed to account for generation kinetics of thermal donors at 450 C, in as-grown and high-temperature preannealed Czochralski silicon crystals. This mechanism is consistent with the main features of the models which have been proposed to explain the formation of oxygen thermal donors in silicon.

  19. Non Heart-Beating Donors in England

    PubMed Central

    Chaib, Eleazar

    2008-01-01

    When transplantation started all organs were retrieved from patients immediately after cardio-respiratory arrest, i.e. from non-heart-beating donors. After the recognition that death resulted from irreversible damage to the brainstem, organ retrieval rapidly switched to patients certified dead after brainstem testing. These heart-beating-donors have become the principal source of organs for transplantation for the last 30 years. The number of heart-beating-donors are declining and this is likely to continue, therefore cadaveric organs from non-heart-beating donor offers a large potential of resources for organ transplantation. The aim of this study is to examine clinical outcomes of non-heart-beating donors in the past 10 years in the UK as an way of decreasing pressure in the huge waiting list for organs transplantation. PMID:18297216

  20. Semen donors' curiosity about donor offspring and the barriers to their knowing.

    PubMed

    Speirs, Jennifer M

    2012-06-01

    The article reports qualitative research findings which explored the meanings of kinship and genetic knowledge of fifteen pre-1990 semen donors in the UK. This is presented in the context of public and academic debates about the regulation of access to genetic information, genetic information as intellectual property and kinship knowledge, and the multiple ownership of genetic information. Semen donors in the UK traditionally were expected to take no interest in what became of their donations and those who did were considered to be unsuitable as donors. However, the present research reveals that men who donated in the past hold varied attitudes, including curiosity about how donor offspring have fared and what they look like. Whilst some donors would welcome direct contact with donor offspring, there are practical and emotional obstacles to satisfying their curiosity. Donors' views reflect the varied understandings in the UK about the implications of genetic relatedness and the time and energy required to maintain and sustain relationships. PMID:22329514

  1. 21 CFR 640.12 - Suitability of donor.

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ... BIOLOGICS ADDITIONAL STANDARDS FOR HUMAN BLOOD AND BLOOD PRODUCTS Red Blood Cells 640.12 Suitability of donor. The source blood for Red Blood Cells shall be obtained from a donor who meets the criteria for donor suitability...

  2. 21 CFR 640.12 - Suitability of donor.

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ... BIOLOGICS ADDITIONAL STANDARDS FOR HUMAN BLOOD AND BLOOD PRODUCTS Red Blood Cells 640.12 Suitability of donor. The source blood for Red Blood Cells shall be obtained from a donor who meets the criteria for donor suitability...

  3. 21 CFR 640.12 - Suitability of donor.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... BIOLOGICS ADDITIONAL STANDARDS FOR HUMAN BLOOD AND BLOOD PRODUCTS Red Blood Cells 640.12 Suitability of donor. The source blood for Red Blood Cells shall be obtained from a donor who meets the criteria for donor suitability...

  4. 21 CFR 660.31 - Suitability of the donor.

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ...STANDARDS FOR DIAGNOSTIC SUBSTANCES FOR LABORATORY TESTS Reagent Red Blood Cells 660.31 Suitability of the donor. Donors of peripheral blood for Reagent Red Blood Cells shall meet the criteria for donor suitability under 640.3 of this...

  5. 21 CFR 660.31 - Suitability of the donor.

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ...STANDARDS FOR DIAGNOSTIC SUBSTANCES FOR LABORATORY TESTS Reagent Red Blood Cells 660.31 Suitability of the donor. Donors of peripheral blood for Reagent Red Blood Cells shall meet the criteria for donor suitability under 640.3 of this...

  6. 21 CFR 640.31 - Suitability of donors.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ...DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) BIOLOGICS ADDITIONAL STANDARDS FOR HUMAN BLOOD AND BLOOD PRODUCTS Plasma 640.31 Suitability of donors. (a) Whole blood donors shall meet the criteria for donor suitability prescribed...

  7. 21 CFR 640.31 - Suitability of donors.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ...DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) BIOLOGICS ADDITIONAL STANDARDS FOR HUMAN BLOOD AND BLOOD PRODUCTS Plasma 640.31 Suitability of donors. (a) Whole blood donors shall meet the criteria for donor suitability prescribed...

  8. 21 CFR 640.31 - Suitability of donors.

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ...DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) BIOLOGICS ADDITIONAL STANDARDS FOR HUMAN BLOOD AND BLOOD PRODUCTS Plasma 640.31 Suitability of donors. (a) Whole blood donors shall meet the criteria for donor suitability prescribed...

  9. 21 CFR 640.31 - Suitability of donors.

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ...DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) BIOLOGICS ADDITIONAL STANDARDS FOR HUMAN BLOOD AND BLOOD PRODUCTS Plasma 640.31 Suitability of donors. (a) Whole blood donors shall meet the criteria for donor suitability prescribed...

  10. 21 CFR 640.31 - Suitability of donors.

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ...DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) BIOLOGICS ADDITIONAL STANDARDS FOR HUMAN BLOOD AND BLOOD PRODUCTS Plasma 640.31 Suitability of donors. (a) Whole blood donors shall meet the criteria for donor suitability prescribed...

  11. 21 CFR 640.12 - Suitability of donor.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... ADDITIONAL STANDARDS FOR HUMAN BLOOD AND BLOOD PRODUCTS Red Blood Cells 640.12 Suitability of donor. The source blood for Red Blood Cells shall be obtained from a donor who meets the criteria for donor suitability prescribed in ...

  12. 21 CFR 660.31 - Suitability of the donor.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ...STANDARDS FOR DIAGNOSTIC SUBSTANCES FOR LABORATORY TESTS Reagent Red Blood Cells 660.31 Suitability of the donor. Donors of peripheral blood for Reagent Red Blood Cells shall meet the criteria for donor suitability under 640.3 of this...

  13. Gamete donors' satisfaction; gender differences and similarities among oocyte and sperm donors in a national sample

    PubMed Central

    Svanberg, Agneta Skoog; Lampic, Claudia; Gejerwall, Ann-Louise; Gudmundsson, Johannes; Karlstr鰉, Per-olof; Solensten, Nils-Gunnar; Sydsj, Gunilla

    2013-01-01

    ObjectiveTo explore oocyte and sperm donors' emotional stress, experiences of care and satisfaction after donation. DesignProspective multicenter study. SettingsAll fertility clinics performing gamete donation in Sweden during the period 2005 to 2008. PopulationOf 220 eligible oocyte donors who were approached, 181 agreed to complete the first questionnaire and 165 completed the second questionnaire 2?months after oocyte donation. Of 156 eligible sperm donors 119 accepted to complete the first questionnaire before donation. Eighty-nine participants completed the second questionnaire 2?months after sperm donation. MethodsStandardized and study-specific questionnaires. Main outcome measuresSatisfaction with the donation, respondents' mental health and overall care. ResultsA larger percentage of sperm donors (97.8%) were satisfied with their overall experience of being a donor than oocyte donors (85.9%, p?=?0.003). Some oocyte and sperm donors did not receive sufficient information about practical issues (9.1% and 13.5%, respectively) and future consequences (12.8% and 3.4%, respectively, p?=?0.014). The donors' symptoms of anxiety and depression did not show any differences in relation to negative or positive perceptions of satisfaction. The donors who did not indicate ambivalence before treatment were on average almost five times more satisfied compared with those who did indicate ambivalence (odds ratio 4.71; 95% CI 1.3416.51). ConclusionsMost donors were satisfied with their contribution after the donation. Oocyte and sperm donors who expressed ambivalence before donation were less satisfied after donation. In vitro fertilization staff fulfilled most of the donors' needs for information and care. Please cite this article as: Skoog Svanberg A, Lampic C, Gejerwall A-L, Gudmundsson J, Karlstr鰉 P-O, Solensten N-G, Sydsj G. Gamete donors satisfaction; gender differences and similarities among oocyte and sperm donors in a national sample. Acta Obstet Gynecol Scand 2013; 92:10491056. PMID:23611727

  14. Donor Research in Australia: Challenges and Promise

    PubMed Central

    Masser, Barbara; Smith, Geoff; Williams, Lisa A.

    2014-01-01

    Summary Donors are the key to the core business of Blood Collection Agencies (BCAs). However, historically, they have not been a focus of research undertaken by these organizations. This model is now changing, with significant donor research groups established in a number of countries, including Australia. Donor research in the Australian Red Cross Blood Service (Blood Service) is concentrated in the Donor and Community Research (DCR) team. Cognizant of the complex and ever-changing landscape with regard to optimal donor management, the DCR team collaborates with academics located at universities around Australia to coordinate a broad program of research that addresses both short- and-long term challenges to the blood supply. This type of collaboration is not, however, without challenges. Two major collaborative programs of the Blood Service's research, focusing on i) the recruitment and retention of plasmapheresis donors and ii) the role of the emotion pride in donor motivation and return, are showcased to elucidate how the challenges of conducting collaborative BCA research can be met. In so doing, these and the other research programs described herein demonstrate how the Blood Service supports and contributes to research that not only revises operational procedures but also contributes to advances in basic science. PMID:25254025

  15. Ultrasensitive photoelectrochemical immunoassay through tag induced exciton trapping.

    PubMed

    Wen, Guangming; Ju, Huangxian

    2015-03-01

    The development of photoelectrochemical (PEC) sensors with novel principles is of significance in realizing sensitive and low-cost detection. This work uses CuO NPs labeled antibody to construct a simple and sensitive sandwich-type immunobiosensor for the detection of protein. The detection signal is produced by dissolving the CuO NPs to release copper ions, which are then added on a quantum dots (QDs) modified F-doped tin oxide to quench the photocurrent of QDs via copper ion-induced formation of exciton trapping. The formed exciton trapping blocks the escape of photoelectron and thus leads to a "signal off" PEC method for sensitive immunoassay. The proposed method shows a detectable range from 0.05 to 500 ng/mL for ?-fetoprotein (AFP) with a detection limit (LOD) of 0.038 ng/mL. This work further extends the application of exciton trapping-based PEC biosensing strategy in bioanalysis. The sensitive analytical performance of the designed route implies a promising potential of the PEC sensing in clinical diagnosis. PMID:25618699

  16. Microfluidic immunoassays as rapid saliva-based clinical diagnostics

    PubMed Central

    Herr, Amy E.; Hatch, Anson V.; Throckmorton, Daniel J.; Tran, Huu M.; Brennan, James S.; Giannobile, William V.; Singh, Anup K.

    2007-01-01

    At present, point-of-care (POC) diagnostics typically provide a binary indication of health status (e.g., home pregnancy test strip). Before anticipatory use of diagnostics for assessment of complex diseases becomes widespread, development of sophisticated bioassays capable of quantitatively measuring disease biomarkers is necessary. Successful translation of new bioassays into clinical settings demands the ability to monitor both the onset and progression of disease. Here we report on a clinical POC diagnostic that enables rapid quantitation of an oral disease biomarker in human saliva by using a monolithic disposable cartridge designed to operate in a compact analytical instrument. Our microfluidic method facilitates hands-free saliva analysis by integrating sample pretreatment (filtering, enrichment, mixing) with electrophoretic immunoassays to quickly measure analyte concentrations in minimally pretreated saliva samples. Using 20 ?l of saliva, we demonstrate rapid (<10 min) measurement of the collagen-cleaving enzyme matrix metalloproteinase-8 (MMP-8) in saliva from healthy and periodontally diseased subjects. In addition to physiologically measurable indicators of periodontal disease, conventional measurements of salivary MMP-8 were used to validate the microfluidic assays described in this proof-of-principle study. The microchip-based POC diagnostic demonstrated is applicable to rapid, reliable measurement of proteinaceous disease biomarkers in biological fluids. PMID:17374724

  17. A Portable Analyzer for Pouch-Actuated, Immunoassay Cassettes

    PubMed Central

    Qiu, Xianbo; Liu, Changchun; Mauk, Michael G.; Hart, Robert W.; Chen, Dafeng; Qiu, Jing; Kientz, Terry; Fiene, Jonathan; Bau, Haim H.

    2011-01-01

    A portable, small footprint, light, general purpose analyzer (processor) to control the flow in immunoassay cassettes and to facilitate the detection of test results is described. The durable analyzer accepts disposable cassettes that contain pouches and reaction chambers for various unit operations such as hydration of dry reagents, stirring, and incubation. The analyzer includes individually controlled, linear actuators to compress the pouches in the cassette, which facilitates the pumping and mixing of sample and reagents, and to close diaphragm-based valves for flow control. The same types of actuators are used to compress pouches and actuate valves. The analyzer also houses a compact OEM scanner/reader to excite fluorescence and detect emission from labels. The analyzer is hydraulically isolated from the cassette, reducing the possibility of cross-contamination. The analyzer facilitates programmable, automated execution of a sequence of operations such as pumping and valving in a timely fashion, reducing the level of expertise required from the operator and the possibility for errors. The analyzer抯 design is modular and expandable to accommodate cassettes of various complexities and additional functionalities. In this paper, the utility of the analyzer has been demonstrated with the execution of a simple, consecutive, lateral flow assay of a model biological system and the test results were detected with up converting phosphor labels that are excited at infrared frequencies and emit in the visible spectrum. PMID:22125359

  18. Sensitive, Fast, and Specific Immunoassays for Methyltestosterone Detection

    PubMed Central

    Kong, Na; Song, Shanshan; Peng, Juan; Liu, Liqiang; Kuang, Hua; Xu, Chuanlai

    2015-01-01

    An indirect competitive enzyme-linked immunosorbent assay (icELISA) and an immunochromatographic strip assay using a highly specific monoclonal antibody, were developed to detect methyltestosterone (MT) residues in animal feed. The optimized icELISA had a half-inhibition concentration value of 0.26 ng/mL and a limit of detection value of 0.045 ng/mL. There was no cross-reactivity with eight analogues, revealing high specificity for MT. Based on icELISA results, the recovery rate of MT in animal feed was 82.4%100.6%. The results were in accordance with those obtained by gas chromatography-mass spectrometry. The developed immunochromatographic strip assay, as the first report for MT detection, had a visual cut-off value of 1 ng/mL in PBS, 2.5 ng/g in fish feed, and 2.5 ng/g in pig feed. Therefore, these immunoassays are useful and fast tools for MT residue detection in animal feed. PMID:25938198

  19. Rapid dioxin screening of milk and water by enzyme immunoassay

    SciTech Connect

    Harrison, R.O.; Carlson, R.E.; Shirkhan, H.

    1995-12-01

    A simple and easy to use enzyme immunoassay (EIA) system has been developed for rapid screening of 2,3,7,8-tetrachlorodibenzo-p-dioxin (2378D). This EIA has been adapted to analysis of water and milk using an automated system for extraction of liquid samples. Water analysis can be performed directly following extraction and solvent exchange with no extract clean-up. The same automated system is used for milk extraction and the extract is then cleaned chromatographically using the automated FMS Dioxin-Prep{trademark} System. Sensitivity for 2378D in the EIA is approximately 100 pg per analysis. Thus sensitivity to 10 ppt 2378D (whole weight basis) in milk is possible using only 50 ml or less of sample and sensitivity to 0.1 ppt 2378D in water is possible using 1-2 liters of sample. Total time for sample preparation and analysis is about 3 hours for water and 4.5 hours for milk.

  20. Backscattering particle immunoassays in wire-guide droplet manipulations

    PubMed Central

    Yoon, Jeong-Yeol; You, David J

    2008-01-01

    A simpler way for manipulating droplets on a flat surface was demonstrated, eliminating the complications in the existing methods of open-surface digital microfluidics. Programmed and motorized movements of 10 ?L droplets were demonstrated using stepper motors and microcontrollers, including merging, complicated movement along the programmed path, and rapid mixing. Latex immunoagglutination assays for mouse immunoglobulin G, bovine viral diarrhea virus and Escherichia coli were demonstrated by merging two droplets on a superhydrophobic surface (contact angle = 155 2) and using subsequent back light scattering detection, with detection limits of 50 pg mL-1, 2.5 TCID50 mL-1 and 85 CFU mL-1, respectively, all significantly lower than the other immunoassay demonstrations in conventional microfluidics (~1 ng mL-1 for proteins, ~100 TCID50 mL-1 for viruses and ~100 CFU mL-1 for bacteria). Advantages of this system over conventional microfluidics or microwell plate assays include: (1) minimized biofouling and repeated use (>100 times) of a platform; (2) possibility of nanoliter droplet manipulation; (3) reprogrammability with a computer or a game pad interface. PMID:19014703

  1. Analysis of cooked-food mutagens by HPLC/immunoassay

    SciTech Connect

    Watkins, B.E.; Vanderlaan, M.; Felton, J.S.

    1988-10-10

    Recognizing that an immunoassay for these aminoimidazoazaarenes (AIAs) could be an analytical method with higher sample throughput and would be less costly, we developed a set of monoclonal antibodies that selectively bind to each of the AIAs. We selected PhIP (6-phenyl-2-amino-1-methylimidazo(4,5-b)pyridine) for the initial assay development because it is the AIA that is most abundant by mass in cooked meat and it is the most genotoxic AIA in mammalian-cell short-term bioassays. It is, however, the least active AIA in the Ames Salmonella mutagenesis assay. We have recently developed a set of four monoclonals that bind to the PhIP. These antibodies were produced by standard methods and were derived from the immunogen described previously. The binding specificity of each of these antibodies has been well characterized. The most specific antibody, called PhIP-1, will bind PhIP with a 50% inhibition point (I/sub 50/) of 30 ng, and it will not bind to any of the other AIA mutagens, nor to any of a number of synthetically produced derivatives of PhIP, such as Iso-PhIP (6-phenyl-2-amino-3-methylimidazo(4,5-b)pyridine). PhIP-1 does bind with 2-deamino-PhIP and 2-deamino-2-nitro-PhIP with an I/sub 50/ of 13 and 16 ng, respectively. 9 refs., 3 figs.

  2. The microassay on a card: A rugged, portable immunoassay

    NASA Technical Reports Server (NTRS)

    Kidwell, David

    1991-01-01

    The Microassay on a Card (MAC) is a portable, hand-held, non-instrumental immunoassay that can test for the presence of a wide variety of substances in the environment. The MAC is a simple device to use. A drop of test solution is placed on one side of the card and within five minutes a color is developed on the other side in proportion to the amount of substance in the test solution, with sensitivity approaching 10 ng/ml. The MAC is self-contained and self-timed; no reagents or timing is necessary. The MAC may be configured with multiple wells to provide simultaneous testing for multiple species. As envisioned, the MAC will be employed first as an on-site screen for drugs of abuse in urine or saliva. If the MAC can be used as a screen of saliva for drugs of abuse, it could be applied to driving while intoxicated, use of drugs on the job, or testing of the identity of seized materials. With appropriate modifications, the MAC also could be used to test for environmental toxins or pollutants.

  3. Designing novel nano-immunoassays: antibody orientation versus sensitivity

    NASA Astrophysics Data System (ADS)

    Puertas, S.; Moros, M.; Fern醤dez-Pacheco, R.; Ibarra, M. R.; Graz, V.; de la Fuente, J. M.

    2010-12-01

    There is a growing interest in the use of magnetic nanoparticles (MNPs) for their application in quantitative and highly sensitive biosensors. Their use as labels of biological recognition events and their detection by means of some magnetic method constitute a very promising strategy for quantitative high-sensitive lateral-flow assays. In this paper, we report the importance of nanoparticle functionalization for the improvement of sensitivity for a lateral-flow immunoassay. More precisely, we have found that immobilization of IgG anti-hCG through its polysaccharide moieties on MNPs allows more successful recognition of the hCG hormone. Although we have used the detection of hCG as a model in this work, the strategy of binding antibodies to MNPs through its sugar chains reported here is applicable to other antibodies. It has huge potential as it will be very useful for the development of quantitative and high-sensitive lateral-flow assays for its use on human and veterinary, medicine, food and beverage manufacturing, pharmaceutical, medical biologics and personal care product production, environmental remediation, etc.

  4. Graphene oxide-based SPR biosensor chip for immunoassay applications

    NASA Astrophysics Data System (ADS)

    Chiu, Nan-Fu; Huang, Teng-Yi; Lai, Hsin-Chih; Liu, Kou-Chen

    2014-08-01

    This work develops a highly sensitive immunoassay sensor for use in graphene oxide sheet (GOS)-based surface plasmon resonance (SPR) chips. This sensing film, which is formed by chemically modifying a GOS surface, has covalent bonds that strongly interact with the bovine serum albumin (BSA), explaining why it has a higher sensitivity. This GOS film-based SPR chip has a BSA concentration detection limit that is 100 times higher than that of the conventional Au-film-based sensor. The affinity constants ( K A) on the GOS film-based SPR chip and the conventional SPR chip for 100 ?g/ml BSA are 80.82 106 M-1 and 15.67 106 M-1, respectively. Therefore, the affinity constant of the GOS film-based SPR chip is 5.2 times higher than that of the conventional chip. With respect to the protein-protein interaction, the SPR sensor capability to detect angle changes at a low concentration anti-BSA of 75.75 nM on the GOS film-based SPR chip and the conventional SPR chip is 36.1867 and 26.1759 mdeg, respectively. At a high concentration, anti-BSA of 378.78 nM on the GOS film-based SPR chip and the conventional SPR chip reveals two times increases in the SPR angle shift. Above results demonstrate that the GOS film is promising for highly sensitive clinical diagnostic applications.

  5. Comparison of different immunoassays for CA 19-9.

    PubMed

    Stern, P; Friedecky, B; Bartos, V; Bezdickova, D; Vavrova, J; Uhrova, J; Rozprimova, L; Zima, T; Palicka, V

    2001-12-01

    We compared six routinely employed immunoassay kits: Architect i2000 and AxSYM, Abbott Laboratories; Elecsys 2010, Roche Diagnostics; ELSA, CIS-BioInternational; Immulite 1, Diagnostic Products Corporation; and IRMA-mat, Byk-Sangtec Diagnostica. Using all analytical systems, we measured identical groups of clinical samples completed with selected control samples. The repeatability of measurements (coefficient of variation) ranged from 2.1% (Elecsys 2010) to 6.7% (ELSA). The parameters of Passing-Bablok regression show significant systematic differences among analytical systems. Data from a Bland-Altman diagram suggest that these differences project onto other, still more significant individual differences among individual samples. Though the cut-off values differ between various systems, no similar clinical efficacy appears to be attained. The behavior of individual systems is quite different for identical control materials and does not necessarily duplicate the calibration for biological samples. The results of determining CA 19-9 cannot be extrapolated from one analytical technique to another, even in cases where the same monoclonal antibody is used. Standardization of CA 19-9 measurement systems is necessary to allow use of the results for the purposes of evidence-based medicine. PMID:11798090

  6. System and method for a parallel immunoassay system

    DOEpatents

    Stevens, Fred J. (Naperville, IL)

    2002-01-01

    A method and system for detecting a target antigen using massively parallel immunoassay technology. In this system, high affinity antibodies of the antigen are covalently linked to small beads or particles. The beads are exposed to a solution containing DNA-oligomer-mimics of the antigen. The mimics which are reactive with the covalently attached antibody or antibodies will bind to the appropriate antibody molecule on the bead. The particles or beads are then washed to remove any unbound DNA-oligomer-mimics and are then immobilized or trapped. The bead-antibody complexes are then exposed to a test solution which may contain the targeted antigens. If the antigen is present it will replace the mimic since it has a greater affinity for the respective antibody. The particles are then removed from the solution leaving a residual solution. This residual solution is applied a DNA chip containing many samples of complimentary DNA. If the DNA tag from a mimic binds with its complimentary DNA, it indicates the presence of the target antigen. A flourescent tag can be used to more easily identify the bound DNA tag.

  7. Interventional radiology in living donor liver transplant

    PubMed Central

    Cheng, Yu-Fan; Ou, Hsin-You; Yu, Chun-Yen; Tsang, Leo Leung-Chit; Huang, Tung-Liang; Chen, Tai-Yi; Hsu, Hsien-Wen; Concerjero, Allan M; Wang, Chih-Chi; Wang, Shih-Ho; Lin, Tsan-Shiun; Liu, Yueh-Wei; Yong, Chee-Chien; Lin, Yu-Hung; Lin, Chih-Che; Chiu, King-Wah; Jawan, Bruno; Eng, Hock-Liew; Chen, Chao-Long

    2014-01-01

    The shortage of deceased donor liver grafts led to the use of living donor liver transplant (LDLT). Patients who undergo LDLT have a higher risk of complications than those who undergo deceased donor liver transplantation (LT). Interventional radiology has acquired a key role in every LT program by treating the majority of vascular and non-vascular post-transplant complications, improving graft and patient survival and avoiding, in the majority of cases, surgical revision and/or re-transplant. The aim of this paper is to review indications, diagnostic modalities, technical considerations, achievements and potential complications of interventional radiology procedures after LDLT. PMID:24876742

  8. Donor-acceptor heteroleptic open sandwiches.

    PubMed

    Merino, Gabriel; Beltr醤, Hiram I; Vela, Alberto

    2006-02-01

    A series of donor-acceptor heteroleptic open sandwiches with formula CpM-M'Pyl (M = B, Al, Ga; M' = Li, Na; Cp = cyclopentadienyl; Pyl = pentadienyl) has been designed in silico using density functional theory. The most stable complexes are those containing boron as a donor atom. A molecular orbital analysis shows that the s character of the lone pair located at the group 13 element is mainly responsible for the complex stabilization. It is also found that the surrounding medium has a similar effect on these sandwiches such as in the "classical" donor-acceptor complexes, showing a decrement in the group 13 element-alkaline metal bond lengths. PMID:16441117

  9. Democrats and Donors: Studying Democratization in Africa

    E-print Network

    Dorman, Sara Rich

    2004-01-01

    Since the late 1980s, political scientists, donors, and development workers in East and Southern Africa have devoted much time and resources to the question of 慸emocratization. Yet, it is not clear how this concept of ...

  10. Ultrafast immunoassays by coupling dielectrophoretic biomarker enrichment in nanoslit channel with electrochemical detection on graphene.

    PubMed

    Sanghavi, Bankim J; Varhue, Walter; Rohani, Ali; Liao, Kuo-Tang; Bazydlo, Lindsay A L; Chou, Chia-Fu; Swami, Nathan S

    2015-12-21

    Heterogeneous immunoassays usually require long incubation times to promote specific target binding and several wash steps to eliminate non-specific binding. Hence, signal saturation is rarely achieved at detection limit levels of analyte, leading to significant errors in analyte quantification due to extreme sensitivity of the signals to incubation time and methodology. The poor binding kinetics of immunoassays at detection limit levels can be alleviated through creating an enriched analyte plug in the vicinity of immobilized capture probes to enable signal saturation at higher levels and at earlier times, due to higher analyte association and its faster replenishment at the binding surface. Herein, we achieve this by coupling frequency-selective dielectrophoretic molecular dam enrichment of the target biomarker in physiological media to capture probes immobilized on graphene-modified surfaces in a nanoslit to enable ultrafast immunoassays with near-instantaneous (<2 minutes) signal saturation at dilute biomarker levels (picomolar) within ultra-low sample volumes (picoliters). This methodology is applied to the detection of Prostate Specific Antigen (PSA) diluted in serum samples, followed by validation against a standard two-step immunoassay using three de-identified patient samples. Based on the ability of dielectrophoretic molecular dam analyte enrichment methods to enable the detection of PSA at 1-5 pg mL(-1) levels within a minute, and the relative insensitivity of the signals to incubation time after the first two minutes, we envision its application for improving the sensitivity of immunoassays and their accuracy at detection limit levels. PMID:26496877

  11. 25OHD analogues and vacuum blood collection tubes dramatically affect the accuracy of automated immunoassays

    PubMed Central

    Yu, Songlin; Cheng, Xinqi; Fang, Huiling; Zhang, Ruiping; Han, Jianhua; Qin, Xuzhen; Cheng, Qian; Su, Wei; Hou, Li抋n; Xia, Liangyu; Qiu, Ling

    2015-01-01

    Variations in vitamin D quantification methods are large, and influences of vitamin D analogues and blood collection methods have not been systematically examined. We evaluated the effects of vitamin D analogues 25OHD2 and 3-epi 25OHD3 and blood collection methods on vitamin D measurement, using five immunoassay systems and liquid chromatography-tandem mass spectrometry (LC-MS/MS). Serum samples (332) were selected from routine vitamin D assay requests, including samples with or without 25OHD2 or 3-epi 25OHD3, and analysed using various immunoassay systems. In samples with no 25OHD2 or 3-epi 25OHD3, all immunoassays correlated well with LC-MS/MS. However, the Siemens system produced a large positive mean bias of 12.5?ng/mL and a poor Kappa value when using tubes with clot activator and gel separator. When 25OHD2 or 3-epi 25OHD3 was present, correlations and clinical agreement decreased for all immunoassays. Serum 25OHD in VACUETTE tubes with gel and clot activator, as measured by the Siemens system, produced significantly higher values than did samples collected in VACUETTE tubes with no additives. Bias decreased and clinical agreement improved significantly when using tubes with no additives. In conclusion, most automated immunoassays showed acceptable correlation and agreement with LC-MS/MS; however, 25OHD analogues and blood collection tubes dramatically affected accuracy. PMID:26420221

  12. Superfund innovative technology evaluation (site) program evaluation report for antox BTX water screen (BTX immunoassay)

    SciTech Connect

    Gerlach, R.W.; White, R.J.; O'Leary, N.F.; Van Emon, J.M.

    1993-06-01

    The results of a demonstration of a portable immunoassay for the detection of benzene, toluene, and xylene(s) (BTX) are described in the report. Seventy-nine field samples were obtained from monitoring wells at several sites with gasoline contaminated ground water. Sample splits were analyzed on-site by the BTX immunoassay and in the laboratory by gas chromatography (GC) using EPA Method 8020. The BTX immunoassay was rapid and simple to use. It performed well in identifying high level contamination and gasoline contaminated samples having BTX concentrations greater than 100 ppb. It did not fully meet the claims of the developer of identifying contamination levels down to 25 ppb BTX. Two field samples determined by GC to have between 25 and 100 ppb BTX failed to be classified correctly by the immunoassay. Results from quality assurance samples with BTX concentrations of 2.5, 25, and 100 ppb also showed that false negative results would be expected at higher than a 5 percent rate when BTX contamination levels were between 25 and 100 ppb. However, for samples with higher BTX levels, the immunoassay gave excellent results. Two field samples yielded false positive results compared to GC values, but these samples showed signs of low-level gasoline contamination.

  13. Curved Oligophenylenes as Donors in Shape-Persistent Donor-Acceptor Macrocycles with Solvatofluorochromic Properties.

    PubMed

    Kuwabara, Takuya; Orii, Jun; Segawa, Yasutomo; Itami, Kenichiro

    2015-08-10

    Many optoelectronic organic materials are based on donor-acceptor (D-A) systems with heteroatom-containing electron donors. Herein, we introduce a new molecular design for all-carbon curved oligoparaphenylenes as donors, which results in the generation of unique shape-persistent D-A macrocycles. Two types of acceptor-inserted cycloparaphenylenes were synthesized. These macrocycles display positive solvatofluorochromic properties owing to their D-A characteristics, which were confirmed by theoretical and electrochemical studies. PMID:26140706

  14. Sperm donors describe the experience of contact with their donor-conceived offspring

    PubMed Central

    Hertz, R.; Nelson, M.K.; Kramer, W.

    2015-01-01

    This study explores the attitudes and experiences of 57 sperm donors who responded to a survey posted online in the United States and indicated that they had had contact with their donor-conceived offspring or the parents of their donor-conceived offspring. On average, 18 years had elapsed since the respondents donated sperm. In the interim between donating and having contact with offspring, most had become curious about their offspring. Most made contact through a bank or online registry. Most respondents had communicated with at least one offspring at least once and most had exchanged photos with offspring. Approximately two-thirds had met in person once; the same proportion had communicated over email or text. Other forms of communication were less common. Almost half of the respondents now considered their donor-conceived offspring to be like a family member. At the same time, donors are respectful of the integrity of the family in which their offspring were raised. Donors with contact are open to having their partners and children know their donor-conceived offspring. Although contact is generally positive, donors report that establishing boundaries and defining the relationship can be very difficult. Some donors also urge those who are thinking of donating to consider the consequences and some suggest avoiding anonymity. There were no significant differences in attitudes and experiences between those who donated anonymously and those who had been identity-release for their offspring when they turned 18. PMID:26175887

  15. How Organ Donors are Different from Non-donors: Responsibility, Barriers, and Religious Involvement.

    PubMed

    Range, Lillian M; Brazda, Geoffrey F

    2015-12-01

    To see if religious involvement, previously linked to various health behaviors, was linked to organ donation, 143 ethnically diverse undergraduates stated whether they were registered donors (53% were), and completed measures of organ donation attitudes and religious involvement. Compared with non-donors, donors reported fewer barriers, more family responsibility, and more willingness to receive donor organs, but were not different in religious involvement. Even in 2014, when being a "good Samaritan" by agreeing to organ donation is as easy as checking one box on a driver's license application, religious involvement does not seem to be a factor in checking this box. PMID:25524413

  16. Selecting suitable solid organ transplant donors: Reducing the risk of donor-transmitted infections

    PubMed Central

    Jr, Christopher S Kovacs; Koval, Christine E; van Duin, David; de Morais, Amanda Guedes; Gonzalez, Blanca E; Avery, Robin K; Mawhorter, Steven D; Brizendine, Kyle D; Cober, Eric D; Miranda, Cyndee; Shrestha, Rabin K; Teixeira, Lucileia; Mossad, Sherif B

    2014-01-01

    Selection of the appropriate donor is essential to a successful allograft recipient outcome for solid organ transplantation. Multiple infectious diseases have been transmitted from the donor to the recipient via transplantation. Donor-transmitted infections cause increased morbidity and mortality to the recipient. In recent years, a series of high-profile transmissions of infections have occurred in organ recipients prompting increased attention on the process of improving the selection of an appropriate donor that balances the shortage of needed allografts with an approach that mitigates the risk of donor-transmitted infection to the recipient. Important advances focused on improving donor screening diagnostics, using previously excluded high-risk donors, and individualizing the selection of allografts to recipients based on their prior infection history are serving to increase the donor pool and improve outcomes after transplant. This article serves to review the relevant literature surrounding this topic and to provide a suggested approach to the selection of an appropriate solid organ transplant donor. PMID:25032095

  17. Replacing antibodies with aptamers in lateral flow immunoassay.

    PubMed

    Chen, Ailiang; Yang, Shuming

    2015-09-15

    Aptamers have been identified against various targets as a type of chemical or nucleic acid ligand by systematic evolution of ligands by exponential enrichment (SELEX) with high sensitivity and specificity. Aptamers show remarkable advantages over antibodies due to the nucleic acid nature and target-induced structure-switching properties and are widely used to design various fluorescent, electrochemical, or colorimetric biosensors. However, the practical applications of aptamer-based sensing and diagnostics are still lagging behind those of antibody-based tests. Lateral flow immunoassay (LFIA) represents a well established and appropriate technology among rapid assays because of its low cost and user-friendliness. The antibody-based platform is utilized to detect numerous targets, but it is always hampered by the antibody preparation time, antibody stability, and effect of modification on the antibody. Seeking alternatives to antibodies is an area of active research and is of tremendous importance. Aptamers are receiving increasing attention in lateral flow applications because of a number of important potential performance advantages. We speculate that aptamer-based LFIA may be one of the first platforms for commercial use of aptamer-based diagnosis. This review first gives an introduction to aptamer including the selection process SELEX with its focus on aptamer advantages over antibodies, and then depicts LFIA with its focus on aptamer opportunities in LFIA over antibodies. Furthermore, we summarize the recent advances in the development of aptamer-based lateral flow biosensing assays with the aim to provide a general guide for the design of aptamer-based lateral flow biosensing assays. PMID:25912679

  18. Comparison of a Time-Resolved Fluorescence Immunoassay and an Enzyme-Linked Immunosorbent Assay for the Analysis of Atrazine

    E-print Network

    Hammock, Bruce D.

    Comparison of a Time-Resolved Fluorescence Immunoassay and an Enzyme-Linked Immunosorbent Assay of California, Davis, California 95616 Immunoassays for atrazine based on a time-resolved fluorescent label and an enzyme label were optimized and utilized to measure atrazine in water. The time-resolved fluorescent

  19. A SERS-based microfluidic immunoassay using an in-situ synthesized gold substrate

    NASA Astrophysics Data System (ADS)

    Fan, Kequan; Wang, Zhuyuan; Wu, Lei; Zong, Shenfei; Cui, Yiping

    2015-05-01

    A sensitive SERS (surface-enhanced Raman scattering)-based immunoassay in microfluidic system has been developed with in-situ synthesis of gold substrate and immune reporter named as 4MBA (4-Mercaptobenzoic acid)-labeled immuno-Ag aggregates. The gold substrate was fabricated simply by introducing the hydrogen tetrachloroaurate (III) trihydrate (HAuCl4) solution to microchannels using a microfluidic pump. It was found that the obtained deposited gold nanoparticles were uniform in size and shape. Then the sandwich immunoassays were performed using the gold substrates based on SERS signals. In the immunoassay, the gold nanoparticles decorated surface was modified with certain antibodies to recognize the specific kind of antigen, which was flowed through the microfluidic channel afterwards. Then 4MBA-labeled immuno-Ag aggregates were employed as the SERS probes to quantitatively detect the antigen. The experimental results showed a good specificity and limit of detection (LOD) about 1 ng/mL.

  20. Simple and specific enzyme immunoassay using monoclonal antibodies for serotyping human rotaviruses.

    PubMed Central

    Coulson, B S; Unicomb, L E; Pitson, G A; Bishop, R F

    1987-01-01

    An enzyme immunoassay for serotyping human rotaviruses in stools and in cell culture was developed. Hyperimmune rabbit antisera to rotaviruses were used as capture antibodies, and rotavirus-neutralizing mouse monoclonal antibodies specific for serotypes 1, 2, 3, and 4 were used as detection reagents. Partial purification of monoclonal antibodies and inclusion of skim milk powder in antibody diluents contributed to assay specificity. The sensitivity of this assay was greater than that of a direct enzyme immunoassay in which rotaviruses of the appropriate serotype were adsorbed directly to the solid phase. When fecal extracts were concentrated threefold, this serotyping enzyme immunoassay was of equal specificity and approached the sensitivity of electron microscopy for rotavirus detection. This assay is simple and rapid and is suitable for serotyping the large numbers of isolates obtained from epidemiological studies and vaccine trials. PMID:3033013

  1. Using cheminformatics to predict cross reactivity of 揹esigner drugs to their currently available immunoassays

    PubMed Central

    2014-01-01

    Background A challenge for drug of abuse testing is presented by 慸esigner drugs, compounds typically discovered by modifications of existing clinical drug classes such as amphetamines and cannabinoids. Drug of abuse screening immunoassays directed at amphetamine or methamphetamine only detect a small subset of designer amphetamine-like drugs, and those immunoassays designed for tetrahydrocannabinol metabolites generally do not cross-react with synthetic cannabinoids lacking the classic cannabinoid chemical backbone. This suggests complexity in understanding how to detect and identify whether a patient has taken a molecule of one class or another, impacting clinical care. Methods Cross-reactivity data from immunoassays specifically targeting designer amphetamine-like and synthetic cannabinoid drugs was collected from multiple published sources, and virtual chemical libraries for molecular similarity analysis were built. The virtual library for synthetic cannabinoid analysis contained a total of 169 structures, while the virtual library for amphetamine-type stimulants contained 288 compounds. Two-dimensional (2D) similarity for each test compound was compared to the target molecule of the immunoassay undergoing analysis. Results 2D similarity differentiated between cross-reactive and non-cross-reactive compounds for immunoassays targeting mephedrone/methcathinone, 3,4-methylenedioxypyrovalerone, benzylpiperazine, mephentermine, and synthetic cannabinoids. Conclusions In this study, we applied 2D molecular similarity analysis to the designer amphetamine-type stimulants and synthetic cannabinoids. Similarity calculations can be used to more efficiently decide which drugs and metabolites should be tested in cross-reactivity studies, as well as to design experiments and potentially predict antigens that would lead to immunoassays with cross reactivity for a broader array of designer drugs. PMID:24851137

  2. Discrepancy between Vitamin D Total Immunoassays due to Various Cross-reactivities

    PubMed Central

    Lee, Jun Hyung; Choi, Jee-Hye; Kweon, Oh Joo

    2015-01-01

    Background The purpose of this study was to find out the cause of discrepancy between various automated immunoassays for 25-hydroxy-vitamin D (25-[OH]D). Methods National Institute of Standards & Technology Standard Reference Material (SRM) 972a is SRM for 25-(OH)D and consists of 4 vials of frozen serum with different concentrations of 25-(OH)D. Each concentration was measured 6 times in 3 different immunoassays: ADVIA Vitamin D Total assay (Siemens Healthcare, Erlangen, Germany), ARCHITECT 25-(OH)D (Abbott Laboratories, Abbott Park, IL, USA), and COBAS Vitamin D Total assay (Roche Diagnostics, Basel, Switzerland). Results When using the certified reference values of SRM 972a as it is, discarding the cross-reactivity of each immunoassay, for ADVIA, the coefficient of determination (R2) as a score of regression analysis was 0.8995 and maximal difference between measured value and certified reference value was 3.6 ng/mL in level 3. The R2 and maximal differences of ARCHITECT were 0.5377 and 6.9 ng/mL, respectively, in level 4. Those of COBAS were 0.3674 and 22.3 ng/mL, respectively, in level 4. When considering cross-reactivities of each immunoassays to various 25-(OH)D metabolites, the ADVIA had R2 and maximal difference of 0.9254 and 3.3 ng/mL, respectively, in level 3. For ARCHITECT, the R2 and maximal differences were 0.7602 and 5.1 ng/mL, respectively, in level 1. Those of COBAS were 0.9284 and 4.9 ng/mL, respectively, in level 1. Conclusions The cause of discrepancies between vitamin D immunoassays was mainly on the difference in cross-reactivities to various vitamin D metabolites. The discrepancies can be considerably decreased by considering cross-reactivities of each immunoassay. PMID:26389085

  3. Recombinant Hepatitis A Virus Antigen: Improved Production and Utility in Diagnostic Immunoassays

    PubMed Central

    LaBrecque, F. D.; LaBrecque, D. R.; Klinzman, D.; Perlman, S.; Cederna, J. B.; Winokur, P. L.; Han, J.-Q.; Stapleton, J. T.

    1998-01-01

    Hepatitis A virus (HAV) immunoassays use cell culture-derived HAV antigen to detect HAV-specific antibodies. The current method of production of HAV antigen in tissue culture is time-consuming and expensive. We previously expressed the HAV open reading frame in recombinant vaccinia viruses (rV-ORF). The recombinant HAV polyprotein was accurately processed and was assembled into subviral particles. These particles were bound by HAV-neutralizing antibodies and were able to elicit antibodies which were detected by commercial immunoassays. The present investigation compared the production of HAV antigen by standard tissue culture methods to the production of HAV antigen with the recombinant vaccinia virus system. In addition, HAV and rV-ORF antigens were assessed for their utility in diagnostic immunoassays. Serum or plasma samples from HAV antibody-positive and antibody-negative individuals were evaluated by immunoassay that used either HAV or rV-ORF antigen. All samples (86 of 86) in which HAV antibody was detected by a commercial enzyme-linked immunosorbent assay (ELISA) also tested positive by the recombinant antigen-based immunoassay (VacRIA). Similarly, all samples (50 of 50) that were HAV antibody negative also tested negative by the VacRIA. The lower limit of detection of HAV antibody was similar among immunoassays with either HAV or rV-ORF antigen. Thus, in the population studied, the sensitivity and specificity of the VacRIA were equivalent to those of the commercial ELISA. Since production of recombinant antigen is faster and less expensive than production of traditional HAV antigen, the development of diagnostic HAV antibody tests with recombinant HAV antigen appears warranted. PMID:9650953

  4. The infrared fingerprint signals of silica nanoparticles and its application in immunoassay

    NASA Astrophysics Data System (ADS)

    Ding, Yadan; Chu, Xueying; Hong, Xia; Zou, Peng; Liu, Yichun

    2012-01-01

    Infrared absorption properties of silica nanoparticles were studied. The transverse optical and the longitudinal optical phonon modes from the silica were proved to be the characteristic spectroscopic fingerprint signals. Based on this, a sandwich-structured immunoassay was performed, and the detection of the analyte (human IgG) was achieved by using biofunctional silica nanoparticles as infrared probes. The immunoassay based on Fourier transform infrared reflection absorption spectroscopy of silica nanoparticles shows significant value for potential applications in many areas, such as biomedicine, food safety, and waste treatment.

  5. A Microfluidic Device for Immunoassay-Based Protein Analysis of Single E. coli Bacteria.

    PubMed

    Stratz, Simone; Dittrich, Petra S

    2015-01-01

    We present a method suitable for quantitative analysis of intracellular proteins, metabolites and secondary messengers of single bacterial cells. The method integrates the concept of immunoassays on a microfluidic device that facilitates single cell trapping and isolating in a small volume of a few tens of picoliters. Combination of the benefits of microfluidic systems for single cell analysis with the high analytical selectivity and sensitivity of immunoassays enables the detection of even low abundant intracellular analytes which occur only at a few hundred copies per bacterium. PMID:26542712

  6. Stool diagnosis of giardiasis using a commercially available enzyme immunoassay to detect Giardia-specific antigen 65 (GSA 65).

    PubMed Central

    Rosoff, J D; Sanders, C A; Sonnad, S S; De Lay, P R; Hadley, W K; Vincenzi, F F; Yajko, D M; O'Hanley, P D

    1989-01-01

    A commercially available enzyme immunoassay for the diagnosis of giardiasis was evaluated in a clinical trial. The ProSpecT/Giardia diagnostic test (Alexon, Inc., Mountain View, Calif.) was compared with the standard ova and parasite (O&P) microscopic examination. Additionally, several widely used stool fixatives and a commonly used transport medium were assessed for compatibility with the immunoassay. A total of 325 stool specimens were collected and used to evaluate assay performance. Of those, 93 specimens were collected from symptomatic Giardia O&P-positive patients and 232 specimens were randomly collected from patients as part of a routine health screening procedure. All 93 Giardia O&P-positive stool specimens were strongly positive by visual and spectrophotometric examination using the immunoassay. Of the 232 randomly collected specimens, 16 were positive by O&P examination and immunoassay, 6 were negative by O&P examination but positive by immunoassay, and 1 was positive by O&P examination and negative by immunoassay. There was substantial supportive evidence that indicated that the six immunoassay-positive, O&P-negative specimens were true-positives. When these six specimens were accepted as true-positives, the immunoassay detected almost 30% more cases of Giardia infection than did O&P examination. Its sensitivity and specificity were 96 and 100%, respectively, while the sensitivity and specificity of O&P examination were 74 and 100%, respectively. The immunoassay also performed well on specimens treated with 10% neutral Formalin, sodium acetate-Formalin fixative, and Cary-Blair transport medium. However, the test was not compatible with polyvinyl alcohol-treated specimens. Overall, the ProSpecT/Giardia test was a sensitive, specific immunoassay which was easy to run and interpret. It offers a simple solution to traditional difficulties encountered in diagnosing Giardia infection. PMID:2674196

  7. The Effect of Donor Age on Corneal Transplantation Outcome: Results of the Cornea Donor Study

    PubMed Central

    2009-01-01

    Objective To determine whether graft survival over a 5-year follow-up period using corneal tissue from donors older than 65 years of age is similar to graft survival using corneas from younger donors. Design Multi-center prospective, double-masked, controlled clinical trial Participants 1090 subjects undergoing corneal transplantation for a moderate risk condition (principally Fuchs dystrophy or pseudophakic corneal edema); 11 subjects with ineligible diagnoses were not included Methods 43 participating eye banks provided corneas from donors in the age range of 12 to 75 with endothelial cell densities of 2300 to 3300 cells/mm2, using a random approach without respect to recipient factors. The 105 participating surgeons at 80 sites were masked to information about the donor cornea including donor age. Surgery and post-operative care were performed according to the surgeons usual routines. Subjects were followed for five years. Main Outcome Measures Graft failure, defined as a regraft or a cloudy cornea that was sufficiently opaque as to compromise vision for a minimum of three consecutive months. Results The 5-year cumulative probability of graft survival was 86% in both the <66.0 donor age group and the ?66.0 donor age group (difference = 0%, upper limit of one-sided 95% confidence interval = 4%). In a statistical model with donor age as a continuous variable, there was not a significant relationship between donor age and outcome (P=0.11). Three graft failures were due to primary donor failure, 8 to uncorrectable refractive error, 48 to graft rejection, 46 to endothelial decompensation (23 of which had a prior, resolved episode of probable or definite graft rejection), and 30 to other causes. The distribution of the causes of graft failure did not differ between donor age groups. Conclusions Five-year graft survival for cornea transplants at moderate risk for failure is similar using corneas from donors ? 66.0 years and donors < 66.0 years. Surgeons and patients now have evidence that corneas comparable in quality to those used in this study from donors through age 75 years are suitable for transplantation. PMID:18387407

  8. Bead-based microfluidic immunoassay for diagnosis of Johne's disease

    SciTech Connect

    Wadhwa, Ashutosh; Foote, Robert; Shaw, Robert W; Eda, Shigetoshi

    2012-01-01

    Microfluidics technology offers a platform for development of point-of-care diagnostic devices for various infectious diseases. In this study, we examined whether serodiagnosis of Johne s disease (JD) can be conducted in a bead-based microfluidic assay system. Magnetic micro-beads were coated with antigens of the causative agent of JD, Mycobacterium avium subsp. paratuberculosis. The antigen-coated beads were incubated with serum samples of JD-positive or negative serum samples and then with a fluorescently-labeled secondary antibody (SAB). To confirm binding of serum antibodies to the antigen, the beads were subjected to flow cytometric analysis. Different conditions (dilutions of serum and SAB, types of SAB, and types of magnetic beads) were optimized for a great degree of differentiation between the JD-negative and JD-positive samples. Using the optimized conditions, we tested a well-classified set of 155 serum samples from JD negative and JD-positive cattle by using the bead-based flow cytometric assay. Of 105 JD-positive samples, 63 samples (60%) showed higher antibody binding levels than a cut-off value determined by using antibody binding levels of JD-negative samples. In contrast, only 43-49 JD-positive samples showed higher antibody binding levels than the cut-off value when the samples were tested by commercially-available immunoassays. Microfluidic assays were performed by magnetically immobilizing a number of beads within a microchannel of a glass microchip and detecting antibody on the collected beads by laser-induced fluorescence. Antigen-coated magnetic beads treated with bovine serum sample and fluorescently-labeled SAB were loaded into a microchannel to measure the fluorescence (reflecting level of antibody binding) on the beads in the microfluidic system. When the results of five bovine serum samples obtained with the system were compared to those obtained with the flow cytometer, a high level of correlation (linear regression, r2 = 0.994) was observed. In a further experiment, we magnetically immobilized antigen-coated beads in a microchannel, reacted the beads with serum and SAB in the channel, and detected antibody binding to the beads in the microfluidic system. A strong antibody binding in JD-positive serum was detected, whereas there was only negligible binding in negative control experiments. Our data suggest that the bead-based microfluidic system may form a basis for development of an on-site serodiagnosis of JD. Key Words: Mycobacterium avium ssp. paratuberculosis, Johne s disease, microfluidics, lab-on-a-chip.

  9. Multiplex serum cytokine immunoassay using nanoplasmonic biosensor microarrays.

    PubMed

    Chen, Pengyu; Chung, Meng Ting; McHugh, Walker; Nidetz, Robert; Li, Yuwei; Fu, Jianping; Cornell, Timothy T; Shanley, Thomas P; Kurabayashi, Katsuo

    2015-04-28

    Precise monitoring of the rapidly changing immune status during the course of a disease requires multiplex analysis of cytokines from frequently sampled human blood. However, the current lack of rapid, multiplex, and low volume assays makes immune monitoring for clinical decision-making (e.g., critically ill patients) impractical. Without such assays, immune monitoring is even virtually impossible for infants and neonates with infectious diseases and/or immune mediated disorders as access to their blood in large quantities is prohibited. Localized surface plasmon resonance (LSPR)-based microfluidic optical biosensing is a promising approach to fill this technical gap as it could potentially permit real-time refractometric detection of biomolecular binding on a metallic nanoparticle surface and sensor miniaturization, both leading to rapid and sample-sparing analyte analysis. Despite this promise, practical implementation of such a microfluidic assay for cytokine biomarker detection in serum samples has not been established primarily due to the limited sensitivity of LSPR biosensing. Here, we developed a high-throughput, label-free, multiarrayed LSPR optical biosensor device with 480 nanoplasmonic sensing spots in microfluidic channel arrays and demonstrated parallel multiplex immunoassays of six cytokines in a complex serum matrix on a single device chip while overcoming technical limitations. The device was fabricated using easy-to-implement, one-step microfluidic patterning and antibody conjugation of gold nanorods (AuNRs). When scanning the scattering light intensity across the microarrays of AuNR ensembles with dark-field imaging optics, our LSPR biosensing technique allowed for high-sensitivity quantitative cytokine measurements at concentrations down to 5-20 pg/mL from a 1 ?L serum sample. Using the nanoplasmonic biosensor microarray device, we demonstrated the ability to monitor the inflammatory responses of infants following cardiopulmonary bypass (CPB) surgery through tracking the time-course variations of their serum cytokines. The whole parallel on-chip assays, which involved the loading, incubation, and washing of samples and reagents, and 10-fold replicated multianalyte detection for each sample using the entire biosensor arrays, were completed within 40 min. PMID:25790830

  10. Using a donor insemination management system.

    PubMed

    Richardson, C A; Dickinson, A S; Barratt, C L; Cooke, I D

    1990-07-01

    A computerized database management system has been developed for an expanding donor insemination service. The programs have been written in dBASEIV language and they will run on a single personal computer (IBM or compatible, with a hard disc) or on a network. The central database enables access to stock, donor and couple details and routine statistics e.g. pregnancy rates, although levels of access to the system are restricted by password control. The central database provides an archival component, with data recorded on a daily basis, a service management component assisting in the routine administration of a donor insemination service, a financial component and a statistical component providing for both local and national statistics. Data may be converted for use in other software packages, e.g. SPSSX, SAS (statistical packages), encouraging data manipulation and enabling specific research questions to be addressed more readily. PMID:2207609

  11. Clinical management of the organ donor.

    PubMed

    Arbour, Richard

    2005-01-01

    There is a critical mismatch between available organs for transplant and acutely or critically ill patients with end-stage organ disease. Patients who may benefit from organ transplantation far outnumber available organs. The causes for this imbalance are multiple. One cause is family refusal to donate. A second cause is nonrecognition or delay in determination of brain death. A third cause is donor loss due to profound cardiopulmonary and metabolic instability consequent to brain-stem herniation and brain death. Family refusal may be addressed by education, public awareness, as well as close attention to social, cultural and ethical issues, and optimal communication with donor families. Brain death may be consequent to traumatic brain injury, ischemic versus hemorrhagic stroke, as well as massive cerebral anoxia/ischemic following cardiac arrest. Nonrecognition or delay in brain death determination may be addressed by clinician education and frequent clinical assessment to detect early stages of brain-stem herniation refractory to aggressive measures for control of intracranial pressure. Donor loss due to profound cardiopulmonary and metabolic instability may be addressed by aggressive, mechanism-based treatment for clinical instability based on affected body system, as well as measures to support metabolic activity at the cellular and tissue level in the brain-dead organ donor. This article explores cerebral physiology related to impending brain death and catastrophic intracranial pressure elevations. In addition, physiologic consequences of brain death are correlated with affected body systems and mechanism-based therapies to support organ function pending transplantation. Ethical/legal issues are explored as related to patient autonomy and optimal family outcomes. Effective family communication, astute clinical assessment, and optimal clinical management of the organ donor are illustrated using a case study approach, highlighting the role of the advanced practice nurse in donor management. PMID:16269899

  12. The ethics of living donor lung transplantation.

    PubMed

    Wells, Winfield J; Barr, Mark L

    2005-11-01

    A constant awareness of the risk to the living donors must be maintained with any living donor organ transplantation program, and comprehensive short- and long-term follow-up should be strongly encouraged to maintain the viability of these potentially life-saving procedures. There has been no perioperative or long-term mortality following lobectomy for living lobar lung transplantation, and perioperative risks associated with donor lobectomy seem to be similar to those seen with standard lung resections. These risks might increase, however, if the procedure is offered on an occasional basis and not within a well-established program. The long-term outcomes and functional effects of lobar donation raise important questions that are unanswered. This has proved difficult to follow closely, because of the fact that many donors live far from the transplant medical center and are reluctant to return for routine follow-up evaluation. The death of a recipient can further exacerbate this situation, because there is reluctance to insist on further routine examinations for a grieving donor. Prospective donors must be informed of the morbidity associated with lobectomy and the potential for mortality, and for potential negative recipient outcomes in regard to life expectancy and quality of life after transplantation. Although cadaveric transplantation must be considered because of the risk to the donors, living lobar lung transplantation should continue to be used under properly selected circumstances. The results reported by the authors' group and others are important if this procedure is to be considered as an option at more pulmonary transplant centers in view of the institutional, regional, and international differences in the philosophic and ethical acceptance of the use of living organ donors for transplantation. The integration of ethical discussion into topics that are relevant and of interest to thoracic surgeons, such as living lung donation, is a recent and welcome event. Many of the clinical situations that thoracic surgeons deal with on a daily basis have important and complex ethical implications, and there has been little training to deal effectively with these issues. This is changing as invited discussions on ethically compelling topics are finding their way into journals and the programs of national meetings. What may be of more importance, however, is the development of an ethics curriculum for those training in the specialty. The core curriculum recommended by the Thoracic Surgical Directors Association (which represents the leadership of the 89 approved residency training programs in the United States) has one lecture pertaining to ethics out of the several hundred offerings in its requisite curriculum. It is hoped that this will change in the near future. PMID:16276816

  13. Silver deposition directed by self-assembled gold nanorods for amplified electrochemical immunoassay.

    PubMed

    Zhang, Hongfang; Ning, Danlei; Ma, Lina; Zheng, Jianbin

    2016-01-01

    A novel electrochemical immunoassay was developed based on the signal amplification strategy of silver deposition directed by gold nanorods (AuNRs), which was in-situ assembled on the sandwich immunocomplex. The superstructure formed by the self-assembly of AuNRs provided abundant active sites for the nucleation of silver nanoparticles. In this pathway, the stripping current of silver was greatly enhanced. Using human immunoglobulin G (HIgG) as a model analyte, the ultrasensitive immunoassay showed a wide linear range of six orders of magnitude from 0.1爁g爉L(-1) to 100爌g爉L(-1), with the low detection limit down to 0.08爁g爉L(-1). The practicality of this electrochemical immunoassay for detection of HIgG in serum was validated with the average recovery of 93.9%. In addition, this enzyme-free immunoassay also has the advantages of acceptable reproducibility and specificity, and thus this immunosensing protocol can be extended to the detection of other low-abundant protein biomarkers. PMID:26703256

  14. Comparison of salivary cortisol as measured by different immunoassays and tandem mass spectrometry.

    PubMed

    Miller, Robert; Plessow, Franziska; Rauh, Manfred; Gr鰏chl, Michael; Kirschbaum, Clemens

    2013-01-01

    Assessing the amount of bioavailable cortisol in saliva with immunoassays and thus sampling an endocrine marker of hypothalamus-pituitary-adrenal axis activity is of major interest in both research and clinical practice. However, absolute cortisol concentrations obtained with different immunoassays (IAs) are barely comparable precluding direct comparison between studies or individuals whenever cortisol analyses were not based on the same IA. The present technical report aims to solve this problem by evaluating the validity of, as well as agreement between the most commonly used immunoassays in psychoneuroendocrinological research (i.e., IBL, DRG, Salimetrics, DSL, and DELFIA) and a reference method (LC-MS/MS) in a sample of 195 saliva specimen covering the whole range of cortisol concentrations in adults. A structural equation modelling framework is applied to decompose systematic assay variance and estimate cortisol reference values, which are adjusted for measurement error and interference of salivary cortisone. Our findings reveal nonlinear relations between IAs and LC-MS/MS, which are discussed in terms of IA cross-reactivity with saliva matrix components. Finally guidelines for converting cortisol concentrations being obtained by these immunoassays into comparable reference values are proposed by providing conversion functions, a conversion table, and an online conversion tool. PMID:22641005

  15. Determination of deoxynivalenol in wheat bran and whole-wheat flour by fluorescence polarization immunoassay

    Technology Transfer Automated Retrieval System (TEKTRAN)

    A rapid and accurate fluorescence polarization (FP) immunoassay has been optimized for the determination of deoxynivalenol (DON) in bran and whole-wheat flour. A preliminary treatment with activated charcoal was used to eliminate the strong matrix effect due to highly colored interfering compounds p...

  16. Metallic nanocrystallites-incorporated ordered mesoporous carbon as labels for a sensitive simultaneous multianalyte electrochemical immunoassay.

    PubMed

    Fang, Yishan; Huang, Xinjian; Zeng, Qiang; Wang, Lishi

    2015-11-15

    This work reports on a facile, novel multianalyte electrochemical immunoassay for simultaneous detection of a-fetoprotein (AFP) and human epidermal growth factor receptor type-2 (HER-2) using metal-containing nanomaterials confined in the ordered mesoporous carbon matrix (OMC-M) as labels. Well-dispersed uniform metallic nanocrystallites incorporated OMC materials were fabricated through a simple, economical, and green preparative strategy toward phenolic resol as a carbon source and metal nitrate as metal sources. The large amount of metallic nanocrystallites loading on the OMC nanomaterials, greatly amplified the detection signals, and the good biocompatibility of carbon nanotubes-chitosan retained excellent stability for the sandwich-type immunoassay. Under optimal experimental conditions, the proposed immunoassay exhibited high sensitivity and selectivity for the detection of analytes, providing a better linear response range from 0.001 to 150 ng/mL for AFP and for HER-2, with a lower limit of detectionof 0.6p g/mL and 0.35 pg/mL (S/N=3), respectively. The immunosensor exhibited convenience, low cost, rapidity, good specificity, acceptable stability and reproducibility. Moreover, satisfactory results were obtained for the determination of AFP and HER-2 in real human serum samples, indicating that the developed immunoassay has the potential to find application in clinical detection of AFP and HER-2 and other tumor markers as an alternative approach. PMID:26046316

  17. Hapten Synthesis and Antibody Development for Polychlorinated Dibenzo-p-dioxin Immunoassays

    E-print Network

    Hammock, Bruce D.

    Hapten Synthesis and Antibody Development for Polychlorinated Dibenzo-p-dioxin Immunoassays James R for the detection of dioxins such as TCDD (2,3,7,8-tetrachlorodibenzo-p-dioxin) by ELISA (enzyme-linked immunosorbent assay). These novel haptens contain unsaturation between the halogenated dibenzo-p-dioxin ring

  18. Immunoassays for the cancer biomarker CA125 based on a large-birefringence nematic liquid-

    E-print Network

    Wu, Shin-Tson

    for gastric cancer patients," J. Cancer Res. Clin. Oncol. 133(7), 471476 (2007). 7. B. Zhang, F. F. CaiImmunoassays for the cancer biomarker CA125 based on a large-birefringence nematic liquid- crystal for the immunodetection. Experiments were performed, targeting at the cancer biomarker CA125. We showed that the larger

  19. Enhanced Sensitive Immunoassay: Noncompetitive Phage Anti-Immune Complex Assay for the Determination of Malachite Green

    E-print Network

    Hammock, Bruce D.

    for the Determination of Malachite Green and Leucomalachite Green Jie-Xian Dong,, Chao Xu, Hong Wang,*, Zhi-Li Xiao ABSTRACT: To develop a more sensitive immunoassay for malachite green (MG) and leucomalachite green (LMG for monitoring food safety. KEYWORDS: malachite green, leucomalachite Green, phage anti-immunocomplex assay

  20. Electrophoretic build-up of multi nanoparticle array for a highly sensitive immunoassay

    E-print Network

    Hammock, Bruce D.

    Electrophoretic build-up of multi nanoparticle array for a highly sensitive immunoassay Jin-Hee Han design of the nanoarray, electrophoretic trapping of a nanoparticle was modeled numerically. The trapping facilitated many breakthroughs in the life sciences by identifying specific gene sequences or protein ana

  1. Magnetic colorimetric immunoassay for human interleukin-6 based on the oxidase activity of ceria spheres.

    PubMed

    Peng, Juan; Guan, Jufang; Yao, Huiqin; Jin, Xiaoyong

    2016-01-01

    A novel magnetic colorimetric immunoassay strategy was designed for sensitive detection of human interleukin-6 (IL-6) using ceria spheres as labels. Ceria spheres showed excellent oxidase activity, which can directly catalyze the oxidation of substrate o-phenylenediamine (OPD) to a stable yellow product, 2,3-diaminophenazine (oxOPD). The absorbance of oxOPD was recorded to reflect the level of IL-6. The relatively mild conditions made the immunoassay strategy more robust, reliable, and easy. A linear relationship between absorbance intensity and the logarithm of IL-6 concentrations was obtained in the range of 0.0001-10爊g爉L(-1) with a detection limit of 0.04爌g爉L(-1) (S/N=3). The colorimetric immunoassay exhibited high sensitivity and specificity for the detection of IL-6. This immunoassay has been successfully applied in the detection of IL-6 in serum samples and can be readily extended toward the on-site monitoring of cancer biomarkers in serum samples. PMID:26416691

  2. Fluorescence polarization immunoassay using IgY antibodies for detection of valnemulin in swine tissue

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Immunoglobulin Y (IgY) is derived from egg yolk and has been identified as a cheap and high-yield immunoreagent. The application of IgY in immunoassay for the detection of chemical contaminants in food samples has rarely been reported. In this work, we describe a rapid and sensitive fluorescence p...

  3. ON-SITE MERCURY ANALYSIS OF SOIL AT HAZARDOUS WASTE SITES BY IMMUNOASSAY AND ASV

    EPA Science Inventory

    Two field methods for Hg, immunoassay and anodic stripping voltammetry (ASV), that can provide onsite results for quick decisions at hazardous waste sites were evaluated. Each method was applied to samples from two Superfund sites that contain high levels of Hg; Sulphur Bank Me...

  4. Rapid Detection of Nivalenol and Deoxynivalenol in Wheat Using Surface Plasmon Resonance Immunoassay

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Surface plasmon resonance immunoassay using a monoclonal antibody was developed to measure nivalenol (NIV) and deoxynivalenol (DON) contamination in wheat. A DON-immobilized sensor chip having high sensitivity and stability was prepared, and an SPR detection procedure was developed. The competitiv...

  5. Microarray Immunoassay for Phenoxybenzoic Acid Using Polymer Encapsulated Eu:Gd2O3

    E-print Network

    Hammock, Bruce D.

    Microarray Immunoassay for Phenoxybenzoic Acid Using Polymer Encapsulated Eu:Gd2O3 Nanoparticles red emission, large Stokes shift, photostable laser-induced fluorescence with a long lifetime (1 ms nanostructures (polymer beads, fluorophore-doped nanoparticles, semiconductor quantum dots, inorganic and organic

  6. Monoclonal antibody-based broad-specificity immunoassay for monitoring organophosphorus pesticides in environmental water samples

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The extensive use of organophosphorus pesticides (OPs) in agriculture and domestic settings can result in widespread water contamination. The development of easy-to-use and rapid-screening immunoassay methods in a class-selective manner is a topic of considerable environmental interest. In this wo...

  7. Heparin interferes with the radioenzymatic and homogeneous enzyme immunoassays for aminoglycosides

    SciTech Connect

    Krogstad, D.J.; Granich, G.G.; Murray, P.R.; Pfaller, M.A.; Valdes, R.

    1981-07-01

    Heparin interferes with measurement of aminoglycosides in serum by biological, radioenzymatic, and homogeneous enzyme immunoassay techniques, but not with radioimmunoassay. At concentrations greater than or equal to 10/sup 5/ and greater than or equal to 3 X 10/sup 6/ USP units/L, respectively, it interferes with the radioenzymatic assay by inhibiting the gentamicin 3-acetyltransferase and kanamycin 6'-acetyltransferase enzymes used in the assay. It interferes with the homogeneous enzyme immunoassays for gentamicin and tobramycin (at concentrations greater than or equal to 10/sup 5/ and greater than or equal to10/sup 4/ USP units/L, respectively), but not with the commercially available homogeneous enzyme immunoassays for other drugs. Heparin interference with the homogeneous enzyme immunoassay for aminoglycosides requires both the heparin polyanion and glucose-6-phosphate dehydrogenase bound to a cationic aminoglycoside. This interference can be reproduced with dextran sulfate (but not dextran), and does not occur with free enzyme (glucose-6-phosphate dehydrogenase) alone. Heparin interference with these two assays and at concentrations that may be present in intravenous infusions or in seriously underfilled blood-collection tubes is described. (JMT)

  8. The measurement of triclosan in water using a magnetic particle enzyme immunoassay

    Technology Transfer Automated Retrieval System (TEKTRAN)

    A sensitive magnetic particle-based immunoassay to determine triclosan (5-chloro-2-(2,4-dichlorophenoxy)phenol) in drinking water and wastewater was developed. Rabbit antiserum was produced by immunizing the rabbit with 6-(5-chloro-2-(2,4-dichlorophenoxy)phenoxy)hexanoic acid-keyhole limpet hemocya...

  9. Immunoassay on Free-Standing Electrospun Dapeng Wu, Daewoo Han, and Andrew J. Steckl*

    E-print Network

    Cincinnati, University of

    -HSA and HSA-FITC as an immunoassay model, a linear detection range from 500 ng/mL down to 1 ng/mL is obtained. Electrospinning is an efficient and simple tool to spray micro/nano polymer fibers from viscous polymer solutions

  10. DEVELOPMENT OF A FLUORESCENT LATEX IMMUNOASSAY FOR DETECTION OF SPECTINOMYCIN ANTIOBIOTIC

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Spectinomycin is an antimicrobial agent used to treat infections caused by Gram negative and positive microorganisms in poultry, swine and non-lactating cattle. There is a need to develop a rapid and sensitive method to detect spectinomycin residues in animal tissues. A latex fluorescent immunoassay...

  11. [GoldMag particle-based chemiluminescence immunoassay for human high sensitive C-reactive protein].

    PubMed

    Guo, Boyang; Ma, Le; Zhang, Mengdan; Yang, Jiangcun; DU, Haiping; Ma, Ting; Yali, Cui

    2015-11-01

    To develop a sensitive, accurate detection method for high sensitive C-reactive protein (hsCRP). Methods With the GoldMag particle as the solid phase carrier, horseradish peroxidase (HRP)-lunimo-H2O2 as the chemiluminescence reaction system, we established a chemiluminescent immunoassay for hsCRP detection. Linear range, sensitivity, precision, accuracy and other indicators were evaluated. hsCRP level of 233 clinical human serum specimens were determined and compared by two different methods: the GoldMag particle-based magnetic chemiluminescence enzyme-linked immunoassay established in this study, and the commercialized scattering immunoturbidimetric assay from Germany SIEMENS. Results The chemiluminescent immunoassay for hsCRP based on GoldMag particle had a good linear relationship between 0.15 mg/L and 25 mg/L (R(2)=0.9937), with the detection limit of 0.076 mg/L. The intra-assay precision was less than 10.00% and the inter-assay precision was less than 15.00%. The average recovery rate for accuracy was 97.80%. In the contrast experiment of 233 clinical human serum specimens, the results obtained using the approach established in this study showed a high correlation and consistency with scattering immunoturbidimetric assay from Germany SIEMENS. Conclusion GoldMag particle-based magnetic chemiluminescence enzyme-linked immunoassay for hsCRP has been successfully developed. PMID:26522353

  12. IMMUNOASSAY METHOD FOR THE DETERMINATION OF PENTACHLOROPHENOL IN SOIL AND SEDIMENT

    EPA Science Inventory

    The journal article describes the use of a prototype immunoassay method for the determination of pentacholorphenol (PCP) in soil and sediment. PCP was used as a pesticide and wood preservative and is not currently available to the general public. The paper stresses the importan...

  13. The detection of cocaine in hair specimens using micro-plate enzyme immunoassay.

    PubMed

    Moore, C; Deitermann, D; Lewis, D; Feeley, B; Niedbala, R S

    1999-05-01

    The analysis of hair for drugs of abuse is becoming increasingly popular and is under consideration by the Division of Health and Human Services as a possible alternative or adjunct to urinalysis in workplace programs. The detection of cocaine in human hair using a commercially available micro-plate enzyme immunoassay is described for the first time. Sample size and incubation time were the major variables in the optimization of the method. In order to validate the procedure, the method was applied to 105 consecutive hair samples routinely received into our laboratory. The samples were simultaneously analyzed by the Micro-Plate immunoassay (EIA), as well as our current fluorescence polarization immunoassay (FPIA) procedure and gas chromatography-mass spectrometry (GC/MS). The sensitivity of the EIA and FPIA assays were 75% and 67.8% respectively; specificity 97.4% and 80.5% respectively; and efficiency 91.4 and 77.1% respectively. The Micro-Plate EIA was shown to be a valid alternative to other immunoassay screening methods for the detection of cocaine in hair by demonstrating increased sensitivity, specificity and efficiency over our current technique. PMID:10408118

  14. AUTOMATED FLOW FLUORESCENT IMMUNOASSAY FOR PART PER TRILLION DETECTION OF THE NEONICOTINOID INSECTICIDE THIAMETHOXAM.

    Technology Transfer Automated Retrieval System (TEKTRAN)

    An ultra sensitive automated flow fluorescent immunoassay was developed using the KinExATM 3000 system for quantitative analysis of the neonicotinoid insecticide thiamethoxam. Five monoclonal antibodies were obtained and screened with a competitive ELISA. One monoclonal antibody designated as E6VI ...

  15. A commercial rapid optical immunoassay detects Streptococcus agalactiae from aquatic cultures and clinical specimens

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The BioStar STREPT B Optical ImmunoAssay (OIA) (BioStar OIA Strep B Assay Kit; Biostar Incorporation; Louisville, CO, USA) was used to identify 32 known group B streptococcus (GBS) isolates of fish, dolphin, bovine, and human origin. Thirteen non-GBS isolates from fish and other animals were test...

  16. Analytica Chimica Acta 444 (2001) 169178 Highly sensitive dioxin immunoassay and its

    E-print Network

    Hammock, Bruce D.

    2001-01-01

    Analytica Chimica Acta 444 (2001) 169178 Highly sensitive dioxin immunoassay and its application Jonesc, Daniel P.Y. Changb, Bruce D. Hammocka, a UCD Cancer Research Center, Department of Entomology Tetrachlorodibenzo-p-dioxin (TCDD) is a well-known highly toxic compound that is present in nearly all components

  17. Competitive chemiluminescent anzyme immunoassay for vitamin B12 analysis in human milk.

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Recent discoveries of matrix interferences by haptocorrin (HC) in human milk and serum show that past analyses of vitamin B12 in samples with high HC content might have been inaccurate (Lildballe et al., 2009; Carmel & Agrawal, 2012). We evaluated two competitive enzyme-binding immunoassays for seru...

  18. An ultrasensitive chemiluminescence immunoassay of chloramphenicol based on gold nanoparticles and magnetic beads.

    PubMed

    Tao, Xiaoqi; Jiang, Haiyang; Yu, Xuezhi; Zhu, Jinghui; Wang, Xia; Wang, Zhanhui; Niu, Lanlan; Wu, Xiaoping; Shen, Jianzhong

    2013-05-01

    A competitive, direct, chemiluminescent immunoassay based on a magnetic beads (MBs) separation and gold nanoparticles (AuNPs) labelling technique to detect chloramphenicol (CAP) has been developed. Horseradish peroxidase (HRP)-labelled anti-CAP monoclonal antibody conjugated with AuNPs and antigen-immobilized MBs were prepared. After optimization parameters of immunocomplex MBs, the IC50 values of chemiluminescence magnetic nanoparticles immunoassay (CL-MBs-nano-immunoassay) were 0.017 礸 L(-1) for extract method I and 0.17 礸 L(-1) for extract method II. The immunoassay with two extract methods was applied to detect CAP in milk. Comparison of these two extract methods showed that extract method I was advantageous in better sensitivity, in which the sensitivity was 10 times compared to that of extract method II, while extract method II was superior in simple operation, suitable for high throughout screen. The recoveries were 86.7-98.0% (extract method I) and 80.0-103.0% (extract method II), and the coefficients of variation (CVs) were all <15%. The satisfactory recovery with both extract methods and high correlation with traditional ELISA kit in milk system confirmed that the immunomagnetic assay based on AuNPs exhibited promising potential in rapid field screening for trace CAP analysis. PMID:23512826

  19. IMMUNOASSAY METHODS FOR MEASURING ATRAZINE AND 3,5,6-TRICHLORO-2-PYRIDINOL IN FOODS

    EPA Science Inventory

    This chapter describes the use of enzyme-linked immunosorbent assay (ELISA) methods for the analysis of two potential environmental contaminants in food sample media, atrazine and 3,5,6-trichloro-2-pyridinol (3,5,6-TCP). Two different immunoassay formats are employed: a magnetic...

  20. Fluorescence polarization immunoassays for rapid, accurate and sensitive determination of mycotoxins

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Fluorescence polarization immunoassay (FPIA) is a type of homogeneous assay. For low molecular weight antigens, such as mycotoxins, it is based on the competition between an unlabeled antigen and its fluorescent-labeled derivative (tracer) for an antigen-specific antibody. The antigen content is det...

  1. Totally laparoscopic living donor right hepatectomy in a donor with trifurcation of bile duct.

    PubMed

    Chen, Kuo-Hsin; Huang, Chun-Chieh; Siow, Tiing-Foong; Chio, U-Chon; Chen, Shian-Dian; Chen, Ying-Da; Lin, Tzu-Chao; Huang, Shu-Yi; Wu, Jiann-Ming; Jeng, Kuo-Shyang

    2016-01-01

    Donor operation in adult living donor liver transplantation is associated with significant postoperative morbidity. To avoid laparotomy wound complications and shorten postoperative recovery, laparoscopic liver graft harvest has been developed recently. However, to determine the cut point of bile duct is challenging. Herein, we report the application of totally laparoscopic approach for right liver graft harvest in a donor with trifurcation of the bile duct. A19-year-old man volunteered for living donation to his father who suffered from hepatitis B virus-related cirrhosis of liver and hepatocellular carcinoma. The graft was 880爉L with a single right hepatic artery and portal vein. The graft to recipient weight ratio was 1.06. The middle hepatic vein was preserved for the donor and the liver remnant was 42.3%. Two branches of middle hepatic veins were > 5爉m in diameter and needed reconstruction with cryopreserved allograft. Ductoplasty using laparoscopic intracorporeal suture technique was done to achieve single orifice of the graft bile duct. The postoperative course was uneventful for the donor. This report adds evidence of the feasibility of pure laparoscopic right donor hepatectomy and describes the necessary steps for bile duct division in donors with trifurcation of bile duct. PMID:26211878

  2. ALTERNATIVE DONORS EXTEND TRANSPLANTATION FOR PATIENTS WITH LYMPHOMA WHO LACK AN HLA MATCHED DONOR

    PubMed Central

    Bachanova, Veronika; Burns, Linda J.; Wang, Tao; Carreras, Jeanette; Gale, Robert Peter; Wiernik, Peter H.; Ballen, Karen K.; Wirk, Baldeep; Munker, Reinhold; Rizzieri, David A.; Chen, Yi-Bin; Gibson, John; Akpek, G鰎g黱; Costa, Luciano J.; Kamble, Rammurti T.; Aljurf, Mahmoud D.; Hsu, Jack W.; Cairo, Mitchell S.; Schouten, Harry C.; Bacher, Ulrike; Savani, Bipin N.; Wingard, John R.; Lazarus, Hillard M.; Laport, Ginna G.; Montoto, Silvia; Maloney, David G.; Smith, Sonali M.; Brunstein, Claudio; Saber, Wael

    2015-01-01

    Alternative donor transplantation is increasingly used for high risk lymphoma patients. We analyzed 1593 transplant recipients (2000 to 2010) and compared transplant outcomes in recipients of 8/8 allele human leukocyte antigen (HLA)-A, -B, -C, and DRB1 matched unrelated donors (MUD; n=1176), 7/8 allele HLA-matched unrelated donors (MMUD; n=275) and umbilical cord blood donors (1 or 2 units UCB; n=142). Adjusted 3-year non-relapse mortality of MMUD (44%) was higher as compared to MUD (35%; p=0.004), but similar to UCB recipients (37%; p=0.19), although UCB had lower rates of neutrophil and platelet recovery compared to unrelated donor groups. With a median follow-up of 55 months, 3-year adjusted cumulative incidence of relapse was lower after MMUD compared with MUD (25% vs 33%, p=0.003) but similar between UCB and MUD (30% vs 33%; p=0.48). In multivariate analysis UCB recipients had lower risks of acute and chronic graft versus host disease compared with adult donor groups (UCB vs MUD: HR=0.68, p=0.05; HR=0.35; p<0.001). Adjusted 3-year overall survival was comparable (43% MUD, 37% MMUD and 41% UCB). Data highlight that patients with lymphoma have acceptable survival after alternative donor transplantation. MMUD and UCB can expand the curative potential of allotransplant to patients who lack suitable HLA-matched sibling or MUD. PMID:25402415

  3. Surface-Enhanced Raman Scattering (SERS) for Detection in Immunoassays: applications, fundamentals, and optimization

    SciTech Connect

    Jeremy Daniel Driskell

    2006-08-09

    Immunoassays have been utilized for the detection of biological analytes for several decades. Many formats and detection strategies have been explored, each having unique advantages and disadvantages. More recently, surface-enhanced Raman scattering (SERS) has been introduced as a readout method for immunoassays, and has shown great potential to meet many key analytical figures of merit. This technology is in its infancy and this dissertation explores the diversity of this method as well as the mechanism responsible for surface enhancement. Approaches to reduce assay times are also investigated. Implementing the knowledge gained from these studies will lead to a more sensitive immunoassay requiring less time than its predecessors. This dissertation is organized into six sections. The first section includes a literature review of the previous work that led to this dissertation. A general overview of the different approaches to immunoassays is given, outlining the strengths and weaknesses of each. Included is a detailed review of binding kinetics, which is central for decreasing assay times. Next, the theoretical underpinnings of SERS is reviewed at its current level of understanding. Past work has argued that surface plasmon resonance (SPR) of the enhancing substrate influences the SERS signal; therefore, the SPR of the extrinsic Raman labels (ERLs) utilized in our SERS-based immunoassay is discussed. Four original research chapters follow the Introduction, each presented as separate manuscripts. Chapter 2 modifies a SERS-based immunoassay previously developed in our group, extending it to the low-level detection of viral pathogens and demonstrating its versatility in terms of analyte type, Chapter 3 investigates the influence of ERL size, material composition, and separation distance between the ERLs and capture substrate on the SERS signal. This chapter links SPR with SERS enhancement factors and is consistent with many of the results from theoretical treatments of SPR and SERS. Chapter 4 introduces a novel method of reducing sample incubation time via capture substrate rotation. Moreover, this work led to a method of virus quantification without the use of standards. Chapter 5 extends the methodology developed in Chapter 4 to both the antigen and ERL labeling step to perform assays with improved analytical performance in less time than can be accomplished in diffusion controlled assays. This dissertation concludes with a general summary and speculates on the future of this exciting approach to carrying out immunoassays.

  4. Compliance with donor age recommendations in oocyte donor recruitment advertisements in the USA.

    PubMed

    Alberta, Hillary B; Berry, Roberta M; Levine, Aaron D

    2013-04-01

    IVF using donated oocytes offers benefits to many infertile patients, yet the technique also raises a number of ethical concerns, including worries about potential physical and psychological risks to oocyte donors. In the USA, oversight of oocyte donation consists of a combination of federal and state regulations and self-regulatory guidelines promulgated by the American Society for Reproductive Medicine. This study assesses compliance with one of these self-regulatory guidelines - specifically, ASRM's preferred minimum age for donors of 21. To assess compliance, 539 oocyte donor recruitment advertisements from two recruitment channels (Craigslist and college newspapers) were collected and evaluated. Of these, 61% in the Craigslist dataset and 43% in the college newspaper dataset listed minimum ages between 18 and 20, which is inconsistent with ASRM's preferred minimum age recommendation of 21. Advertisements placed by oocyte donor recruitment agencies were more likely than advertisements placed by clinics to specify minimum ages between 18 and 20. These results indicate that ASRM should evaluate and consider revising its donor age guidelines. IVF using donated human eggs can help many patients who have difficulty having children. However, the technique also raises ethical concerns, including concerns about potential physical and psychological harms to egg donors. In the USA, oversight of egg donation relies on a combination of federal and state regulation and professional self-regulation. Governmental regulations address only limited aspects of egg donation, such as the potential spread of infectious diseases and the reporting of success rates, leaving voluntary guidelines developed by an association of medical professionals to address most issues, including ethical concerns raised by the practice. One of these voluntary guidelines recommends that egg donors should be at least 21 years of age. In this article, we analysed 539 egg donor recruitment advertisements published on Craigslist and in college newspapers to see whether fertility clinics and egg donor recruitment agencies follow this recommendation. We found that 61% of advertisements in the Craigslist dataset and 43% of advertisements in the college newspaper dataset listed minimum ages between 18 and 20 and, thus, did not follow the recommendation that egg donors be at least 21 years of age. Advertisements placed by egg donor recruitment agencies were more likely than advertisements placed by fertility clinics to list minimum ages between 18 and 20. These results indicate that the American Society for Reproductive Medicine should evaluate and consider revising its donor age guidelines. PMID:23337419

  5. Functionalized gold nanoclusters as fluorescent labels for immunoassays: Application to human serum immunoglobulin E determination.

    PubMed

    Alonso, Mar韆 Cruz; Trapiella-Alfonso, Laura; Fern醤dez, Jos M Costa; Pereiro, Rosario; Sanz-Medel, Alfredo

    2016-03-15

    A quantitative immunoassay for the determination of immunoglobulin E (IgE) in human serum using gold nanoclusters (AuNCs) as fluorescent label was developed. Water soluble AuNCs were synthesized using lipoic acid and then thoroughly characterized. The obtained AuNCs have a particle size of 2.70.1nm and maximum fluorescence emission at 710nm. The synthesized AuNCs showed very good stability of the fluorescent signal with light exposure and at neutral and slightly basic media. A covalent bioconjugation of these AuNCs with the desired antibody was carried out by the carbodiimide reaction. After due optimization of such bioconjugation reaction, a molar ratio 1:3 (antibody:AuNCs) was selected. The bioconjugate maintained an intense luminescence emission, slightly red-shifted as compared to the free AuNCs. Two typical immunoassay configurations, competitive and sandwich, were assayed and their performance for IgE determination critically compared. After the different immunoassay steps were accomplished, the fluorescence emission of the bioconjugate was measured. While the sandwich format provided a detection limit (DL) of 10ng/mL and a linear range between 25 and 565ng/mL of IgE, the competitive format revealed a DL of 0.2ng/mL with a linear range between 0.3 and 7.1ng/mL The applicability of the more sensitive competitive fluorescent immunoassay was assessed by successful analysis of the IgE in human serum and comparison of results with those from a commercial kit. The main advantages of the proposed AuNCs-based fluorimetric method include a low DL and a simple immunoassay protocol involving few reagents. PMID:26547433

  6. 21 CFR 610.41 - Donor deferral.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ...610.47 when a donor tests reactive by a screening test for HIV or HCV required under 610.40(a) and (b), or when you...other reliable test results or information indicating evidence of HIV or HCV infection. [66 FR 31164, June 11, 2001, as...

  7. 21 CFR 610.41 - Donor deferral.

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ...610.47 when a donor tests reactive by a screening test for HIV or HCV required under 610.40(a) and (b), or when you...other reliable test results or information indicating evidence of HIV or HCV infection. [66 FR 31164, June 11, 2001, as...

  8. 21 CFR 610.41 - Donor deferral.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ...610.47 when a donor tests reactive by a screening test for HIV or HCV required under 610.40(a) and (b), or when you...other reliable test results or information indicating evidence of HIV or HCV infection. [66 FR 31164, June 11, 2001, as...

  9. 21 CFR 610.41 - Donor deferral.

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ...610.47 when a donor tests reactive by a screening test for HIV or HCV required under 610.40(a) and (b), or when you...other reliable test results or information indicating evidence of HIV or HCV infection. [66 FR 31164, June 11, 2001, as...

  10. 21 CFR 610.41 - Donor deferral.

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ...610.47 when a donor tests reactive by a screening test for HIV or HCV required under 610.40(a) and (b), or when you...other reliable test results or information indicating evidence of HIV or HCV infection. [66 FR 31164, June 11, 2001, as...

  11. Do clinicians benefit from gamete donor anonymity?

    PubMed

    Haimes, E V

    1993-09-01

    Two groups of participants are frequently omitted from discussions and studies of donor anonymity in assisted conception: the children conceived and the clinicians providing the service. Past secrecy explains the absence of the children's views, but the absence of a systematic consideration of the clinician's views is more puzzling. Evidence from the history of donor insemination suggests that clinicians have supported keeping such practices secret, not just for the protection of donors, recipients and resultant children but also to protect their own position from the detailed scrutiny of others who had expressed doubts about the practice. However, the various important developments in both the practice and the regulation of assisted conception in the 1980s and the early 1990s may well have alleviated such earlier anxieties. None the less, a growing willingness by clinicians to consider greater openness in gamete donation may be counter-balanced by the nature of their relationship with recipients, the majority of whom still appear to favour secrecy, and by the wider cultural uncertainty about the physiological and symbolic importance of genetic relationships in the development of the individual. It is concluded therefore that future studies of donor anonymity should include clinicians, in order to explore these questions in detail. PMID:8253945

  12. Hydrogen-donor coal liquefaction process

    DOEpatents

    Wilson, Jr., Edward L. (Baytown, TX); Mitchell, Willard N. (Baytown, TX)

    1980-01-01

    Improved liquid yields are obtained during the hydrogen-donor solvent liquefaction of coal and similar carbonaceous solids by maintaining a higher concentration of material having hydrogenation catalytic activity in the downstream section of the liquefaction reactor system than in the upstream section of the system.

  13. 21 CFR 610.41 - Donor deferral.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ...) subsequently may donate Source Plasma for use in the preparation of Hepatitis B Immune Globulin (Human... for anti-HBc or for evidence of infection due to HTLV, types I and II, may serve as a donor of Source....40(a) and (b), or when you are aware of other reliable test results or information...

  14. 21 CFR 610.41 - Donor deferral.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... infection due to hepatitis B surface antigen (HBsAg) when previously tested under 610.40(a), (b), and (e... deferral. (a) You, an establishment that collects human blood or blood components, must defer donors testing reactive by a screening test for evidence of infection due to a communicable disease...

  15. Achieving donor repetition and motivation by block leaders among current blood donors.

    PubMed

    Mart韓-Santana, Josefa D; Beerli-Palacio, Asunci髇

    2012-12-01

    This paper presents an explicative model on the recommendation of donating blood made to relatives and friends by current donors. This model establishes satisfaction and intention to return as direct antecedents, and the quality perceived in the donation process and the existence of inhibitors as indirect antecedents. The results show that (1) the perceived quality has a positive influence on satisfaction and intention to return; (2) the intention to donate again depends positively on satisfaction, but negatively on the existence of internal and external inhibitors; and lastly (3) the recommendation to donate depends on donor satisfaction and their intention to return to donate, this being the most influential factor. At the same time, we contrasted how the model does not vary, whether it is a first-time donor or a repeat donor. PMID:22683233

  16. 21 CFR 640.12 - Suitability of donor.

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ... ADDITIONAL STANDARDS FOR HUMAN BLOOD AND BLOOD PRODUCTS Red Blood Cells 640.12 Suitability of donor. The source blood for Red Blood Cells shall be obtained from a donor who meets the criteria for...

  17. 21 CFR 660.31 - Suitability of the donor.

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ...) BIOLOGICS ADDITIONAL STANDARDS FOR DIAGNOSTIC SUBSTANCES FOR LABORATORY TESTS Reagent Red Blood Cells 660.31 Suitability of the donor. Donors of peripheral blood for Reagent Red Blood Cells shall meet...

  18. 21 CFR 640.12 - Suitability of donor.

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ... ADDITIONAL STANDARDS FOR HUMAN BLOOD AND BLOOD PRODUCTS Red Blood Cells 640.12 Suitability of donor. The source blood for Red Blood Cells shall be obtained from a donor who meets the criteria for...

  19. 21 CFR 660.31 - Suitability of the donor.

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ...) BIOLOGICS ADDITIONAL STANDARDS FOR DIAGNOSTIC SUBSTANCES FOR LABORATORY TESTS Reagent Red Blood Cells 660.31 Suitability of the donor. Donors of peripheral blood for Reagent Red Blood Cells shall meet...

  20. 21 CFR 640.12 - Suitability of donor.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... ADDITIONAL STANDARDS FOR HUMAN BLOOD AND BLOOD PRODUCTS Red Blood Cells 640.12 Suitability of donor. The source blood for Red Blood Cells shall be obtained from a donor who meets the criteria for...

  1. 21 CFR 640.12 - Suitability of donor.

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ... ADDITIONAL STANDARDS FOR HUMAN BLOOD AND BLOOD PRODUCTS Red Blood Cells 640.12 Suitability of donor. The source blood for Red Blood Cells shall be obtained from a donor who meets the criteria for...

  2. 21 CFR 660.31 - Suitability of the donor.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ...) BIOLOGICS ADDITIONAL STANDARDS FOR DIAGNOSTIC SUBSTANCES FOR LABORATORY TESTS Reagent Red Blood Cells 660.31 Suitability of the donor. Donors of peripheral blood for Reagent Red Blood Cells shall meet...

  3. 21 CFR 660.31 - Suitability of the donor.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ...) BIOLOGICS ADDITIONAL STANDARDS FOR DIAGNOSTIC SUBSTANCES FOR LABORATORY TESTS Reagent Red Blood Cells 660.31 Suitability of the donor. Donors of peripheral blood for Reagent Red Blood Cells shall meet...

  4. 21 CFR 640.12 - Suitability of donor.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... ADDITIONAL STANDARDS FOR HUMAN BLOOD AND BLOOD PRODUCTS Red Blood Cells 640.12 Suitability of donor. The source blood for Red Blood Cells shall be obtained from a donor who meets the criteria for...

  5. 21 CFR 660.31 - Suitability of the donor.

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ...) BIOLOGICS ADDITIONAL STANDARDS FOR DIAGNOSTIC SUBSTANCES FOR LABORATORY TESTS Reagent Red Blood Cells 660.31 Suitability of the donor. Donors of peripheral blood for Reagent Red Blood Cells shall meet...

  6. Good Results from IVF Egg Donors Over Age 35

    MedlinePLUS

    ... news/fullstory_155243.html Good Results From IVF Egg Donors Over Age 35 Pregnancy, live birth rates ... Results of in vitro fertilization (IVF) cycles using eggs from older donors are as good as those ...

  7. Alternative donor transplant of benign primary hematologic disorders

    PubMed Central

    Tolar, J; Sodani, P; Symons, H

    2015-01-01

    Hematopoietic SCT is currently the only curative therapy for a range of benign inherited and acquired primary hematologic disorders in children, including BM failure syndromes and hemoglobinopathies. The preferred HLA-matched sibling donor is available for only about 25% of such children. However, there has been substantial progress over the last four decades in the use of alternative donors for those without a matched sibling梚ncluding HLA-matched unrelated donors, HLA-haploidentical related donors and unrelated-donor umbilical cord blood梥o that it is now possible to find a donor for almost every child requiring an allograft. Below, we summarize the relative merits and limitations of the different alternative donors for benign hematologic conditions, first generally, and then in relation to specific disorders, and suggest recommendations for selecting such an alternative donor. PMID:25665040

  8. 21 CFR 640.66 - Immunization of donors.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ...DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) BIOLOGICS ADDITIONAL STANDARDS FOR HUMAN BLOOD AND BLOOD PRODUCTS Source Plasma 640.66 Immunization of donors. If specific immunization of a donor is to be performed, the selection...

  9. 21 CFR 640.73 - Reporting of fatal donor reactions.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ...DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) BIOLOGICS ADDITIONAL STANDARDS FOR HUMAN BLOOD AND BLOOD PRODUCTS Source Plasma 640.73 Reporting of fatal donor reactions. If a donor has a fatal reaction which, in any way, may be...

  10. 21 CFR 640.66 - Immunization of donors.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ...DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) BIOLOGICS ADDITIONAL STANDARDS FOR HUMAN BLOOD AND BLOOD PRODUCTS Source Plasma 640.66 Immunization of donors. If specific immunization of a donor is to be performed, the selection...

  11. 21 CFR 640.73 - Reporting of fatal donor reactions.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ...DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) BIOLOGICS ADDITIONAL STANDARDS FOR HUMAN BLOOD AND BLOOD PRODUCTS Source Plasma 640.73 Reporting of fatal donor reactions. If a donor has a fatal reaction which, in any way, may be...

  12. 21 CFR 640.66 - Immunization of donors.

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ...DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) BIOLOGICS ADDITIONAL STANDARDS FOR HUMAN BLOOD AND BLOOD PRODUCTS Source Plasma 640.66 Immunization of donors. If specific immunization of a donor is to be performed, the selection...

  13. 21 CFR 640.73 - Reporting of fatal donor reactions.

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ...DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) BIOLOGICS ADDITIONAL STANDARDS FOR HUMAN BLOOD AND BLOOD PRODUCTS Source Plasma 640.73 Reporting of fatal donor reactions. If a donor has a fatal reaction which, in any way, may be...

  14. 21 CFR 640.66 - Immunization of donors.

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ...DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) BIOLOGICS ADDITIONAL STANDARDS FOR HUMAN BLOOD AND BLOOD PRODUCTS Source Plasma 640.66 Immunization of donors. If specific immunization of a donor is to be performed, the selection...

  15. 21 CFR 640.73 - Reporting of fatal donor reactions.

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ...DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) BIOLOGICS ADDITIONAL STANDARDS FOR HUMAN BLOOD AND BLOOD PRODUCTS Source Plasma 640.73 Reporting of fatal donor reactions. If a donor has a fatal reaction which, in any way, may be...

  16. 21 CFR 640.73 - Reporting of fatal donor reactions.

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ...DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) BIOLOGICS ADDITIONAL STANDARDS FOR HUMAN BLOOD AND BLOOD PRODUCTS Source Plasma 640.73 Reporting of fatal donor reactions. If a donor has a fatal reaction which, in any way, may be...

  17. 21 CFR 640.66 - Immunization of donors.

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ...DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) BIOLOGICS ADDITIONAL STANDARDS FOR HUMAN BLOOD AND BLOOD PRODUCTS Source Plasma 640.66 Immunization of donors. If specific immunization of a donor is to be performed, the selection...

  18. 21 CFR 640.21 - Suitability of donors.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ...SERVICES (CONTINUED) BIOLOGICS ADDITIONAL STANDARDS FOR HUMAN BLOOD AND BLOOD PRODUCTS Platelets 640.21 Suitability of donors. (a) Whole blood donors shall meet the criteria for suitability prescribed in 640.3....

  19. Donor factors predicting recipient survival after liver retransplantation: the retransplant donor risk index.

    PubMed

    Northup, P G; Pruett, T L; Kashmer, D M; Argo, C K; Berg, C L; Schmitt, T M

    2007-08-01

    The use of extended criteria liver donors (ECD) is controversial, especially in the setting of retransplantation. The aims of this study are to investigate the effects of ECD grafts on retransplantation and to develop a predictive mortality index in liver retransplantation based on the previously established donor risk index. The United Network for Organ Sharing (UNOS) liver transplant dataset was analyzed for all adult, non-status 1, liver retransplantations occurring in the United States since February 2002. All donors were categorized for multiple characteristics of ECD, and using multivariate survival models a retransplant donor risk index (ReTxDRI) was developed. A total of 1327 retransplants were analyzed. There were 611 (46%) recipients who received livers with at least one ECD criterion. The use of ECD grafts in recipients with HCV did not incur worse survival than the non-ECD grafts. The addition of the cause of recipient graft failure to the donor risk index formed the ReTxDRI. After adjusting for multiple recipient factors, the ReTxDRI was predictive of overall recipient survival and was a strongly independent predictor of death after retransplantation (HR 2.49, 95% CI 1.89-3.27, p < 0.0001). The use of the ReTxDRI can improve recipient and donor matching and help to optimize posttransplant survival in liver retransplantation. PMID:17617863

  20. 21 CFR 640.63 - Suitability of donor.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... ADDITIONAL STANDARDS FOR HUMAN BLOOD AND BLOOD PRODUCTS Source Plasma 640.63 Suitability of donor. (a) Method of determining. The suitability of a donor for Source Plasma shall be determined by a qualified... year. (2)(i) A donor who is to be immunized for the production of high-titer plasma shall be...

  1. 21 CFR 640.63 - Suitability of donor.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... ADDITIONAL STANDARDS FOR HUMAN BLOOD AND BLOOD PRODUCTS Source Plasma 640.63 Suitability of donor. (a) Method of determining. The suitability of a donor for Source Plasma shall be determined by a qualified... year. (2)(i) A donor who is to be immunized for the production of high-titer plasma shall be...

  2. 21 CFR 640.63 - Suitability of donor.

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ... ADDITIONAL STANDARDS FOR HUMAN BLOOD AND BLOOD PRODUCTS Source Plasma 640.63 Suitability of donor. (a) Method of determining. The suitability of a donor for Source Plasma shall be determined by a qualified... year. (2)(i) A donor who is to be immunized for the production of high-titer plasma shall be...

  3. 21 CFR 640.63 - Suitability of donor.

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ... ADDITIONAL STANDARDS FOR HUMAN BLOOD AND BLOOD PRODUCTS Source Plasma 640.63 Suitability of donor. (a) Method of determining. The suitability of a donor for Source Plasma shall be determined by a qualified... year. (2)(i) A donor who is to be immunized for the production of high-titer plasma shall be...

  4. 21 CFR 640.63 - Suitability of donor.

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ... ADDITIONAL STANDARDS FOR HUMAN BLOOD AND BLOOD PRODUCTS Source Plasma 640.63 Suitability of donor. (a) Method of determining. The suitability of a donor for Source Plasma shall be determined by a qualified... year. (2)(i) A donor who is to be immunized for the production of high-titer plasma shall be...

  5. Mating by proxy: a novel perspective to donor conception.

    PubMed

    Rodino, Iolanda S; Burton, Peter J; Sanders, Katherine A

    2011-10-01

    How single, partnered lesbian, and partnered heterosexual women undertaking donor insemination rate the importance of donor characteristics is explored in the context of Trivers's parental investment theory. Consistent with this theory, single women placed higher value on biographical traits reflective of the donor's level of potential resources (occupation, hobbies, age) and good character compared with either partnered lesbian or heterosexual women. PMID:21821248

  6. A legal defense for compensating research egg donors.

    PubMed

    Crockin, Susan L

    2010-02-01

    Given the continued need for human eggs for hESCs, this article analyzes and refutes the legal theories against compensating research egg donors, contrasts the legal histories of compensating reproductive donors and human subjects with noncompensation for ESC donors, and suggests that limited compensation is legally defensible. PMID:20144783

  7. Synthesis of a photocontrollable hydrogen sulfide donor using ketoprofenate photocages.

    PubMed

    Fukushima, Naoki; Ieda, Naoya; Sasakura, Kiyoshi; Nagano, Tetsuo; Hanaoka, Kenjiro; Suzuki, Takayoshi; Miyata, Naoki; Nakagawa, Hidehiko

    2014-01-18

    We report the design, synthesis and application of a directly photocontrollable hydrogen sulfide (H2S) donor, which releases H2S proportionally to the intensity and duration of photoirradiation. Photocontrolled H2S release from this donor was also demonstrated in bovine serum. This H2S donor should be suitable for use in various biological systems. PMID:24280741

  8. Development and Application of a Gel-Based Immunoassay for the Rapid Screening of Salbutamol and Ractopamine Residues in Pork.

    PubMed

    Li, Chenglong; Li, Jingya; Jiang, Wenxiao; Zhang, Suxia; Shen, Jianzhong; Wen, Kai; Wang, Zhanhui

    2015-12-01

    Salbutamol (SAL) and ractopamine (RAC) have been illegally used to promote protein synthesis and to increase the feed conversion rate in livestock. However, the residues of SAL and RAC could cause potential hazards for human health. The Ministry of Agriculture of China banned the use of SAL and RAC as growth promoters. In this paper, we provide detailed information on developing a rapid and sensitive gel-based immunoassay for on-site screening of SAL and RAC residues in pork. The detection time was shortened to 20 min. The limits of detection were 0.5 ?g/kg for both SAL and RAC by visual detection, whereas the quantitative gel-based immunoassay enabled the detection of SAL (0.051 ?g/kg) and RAC (0.020 ?g/kg) in spiked pork samples. The gel-based immunoassay showed promise as a multiplexed immunoassay for on-site surveilling of SAL and RAC residues in pork. PMID:26595169

  9. DEMONSTRATION BULLETIN: PCP IMMUNOASSAY TECHNOLOGIES - PENTA RISC BY ENSYS INC., PENTA RAPID BY OHMICRON CORP., ENVIROGARD BY MILLIPORE

    EPA Science Inventory

    The objectives of this demonstration were to test these field screening technologies for accuracy and precision in detecting Pentachlorophenol (PCP) levels in soil and water by comparing their results with those of a confirmatory laboratory. The three immunoassay technologies ...

  10. Hydrophilic, bright CuInS2 quantum dots as Cd-free fluorescent labels in quantitative immunoassay.

    PubMed

    Speranskaya, Elena S; Beloglazova, Natalia V; Ab, Sofie; Aubert, Tangi; Smet, Philippe F; Poelman, Dirk; Goryacheva, Irina Y; De Saeger, Sarah; Hens, Zeger

    2014-07-01

    We report on the synthesis of core-shell CuInS2/ZnS quantum dots (QDs) in organic solution, their encapsulation with a PEG-containing amphiphilic polymer, and the application of the resulting water-soluble QDs as fluorescent label in quantitative immunoassay. By optimizing the methods for core synthesis and shell growth, CuInS2/ZnS QDs were obtained with a quantum yield of 50% on average after hydrophilization. After conjugation with an aflatoxin B1-protein derivative, the obtained QDs were used as fluorescent labels in microplate immunoassay for the quantitative determination of the mycotoxin aflatoxin B1. QDs-based immunoassay showed higher sensitivity compared to enzyme-based immunoassay. PMID:24892375

  11. 21 CFR 1271.50 - How do I determine whether a donor is eligible?

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ... HUMAN CELLS, TISSUES, AND CELLULAR AND TISSUE-BASED PRODUCTS Donor Eligibility 1271.50 How do I... document the eligibility of a cell or tissue donor. (b) Eligible donor. A donor is eligible under...

  12. 21 CFR 1271.50 - How do I determine whether a donor is eligible?

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... HUMAN CELLS, TISSUES, AND CELLULAR AND TISSUE-BASED PRODUCTS Donor Eligibility 1271.50 How do I... document the eligibility of a cell or tissue donor. (b) Eligible donor. A donor is eligible under...

  13. 21 CFR 1271.50 - How do I determine whether a donor is eligible?

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... HUMAN CELLS, TISSUES, AND CELLULAR AND TISSUE-BASED PRODUCTS Donor Eligibility 1271.50 How do I... document the eligibility of a cell or tissue donor. (b) Eligible donor. A donor is eligible under...

  14. 21 CFR 1271.50 - How do I determine whether a donor is eligible?

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ...ACTS ADMINISTERED BY THE FOOD AND DRUG ADMINISTRATION HUMAN CELLS, TISSUES, AND CELLULAR AND TISSUE-BASED PRODUCTS Donor...3(t), must determine and document the eligibility of a cell or tissue donor. (b) Eligible donor . A donor is...

  15. 21 CFR 1271.50 - How do I determine whether a donor is eligible?

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ...ACTS ADMINISTERED BY THE FOOD AND DRUG ADMINISTRATION HUMAN CELLS, TISSUES, AND CELLULAR AND TISSUE-BASED PRODUCTS Donor...3(t), must determine and document the eligibility of a cell or tissue donor. (b) Eligible donor . A donor is...

  16. 21 CFR 1271.50 - How do I determine whether a donor is eligible?

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ...ACTS ADMINISTERED BY THE FOOD AND DRUG ADMINISTRATION HUMAN CELLS, TISSUES, AND CELLULAR AND TISSUE-BASED PRODUCTS Donor...3(t), must determine and document the eligibility of a cell or tissue donor. (b) Eligible donor. A donor is...

  17. 78 FR 66366 - Draft Guidance for Industry: Use of Donor Screening Tests To Test Donors of Human Cells, Tissues...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2013-11-05

    ... enforcement discretion. In the Federal Register of February 28, 2007 (72 FR 9007), FDA announced the... Test Donors of Human Cells, Tissues, and Cellular and Tissue-Based Products for Infection With... entitled ``Guidance for Industry: Use of Donor Screening Tests to Test Donors of Human Cells, Tissues,...

  18. 78 FR 66366 - Draft Guidance for Industry: Use of Donor Screening Tests To Test Donors of Human Cells, Tissues...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2013-11-05

    ...Screening Tests To Test Donors of Human Cells, Tissues, and Cellular and Tissue-Based...Screening Tests to Test Donors of Human Cells, Tissues, and Cellular and Tissue-Based...Eligibility Determination for Donors of Human Cells, Tissues, and Cellular and...

  19. Diet and Asthma: Vitamins and Methyl Donors

    PubMed Central

    Han, Yueh-Ying; Blatter, Josh; Brehm, John M.; Forno, Erick; Litonjua, Augusto A; Celed髇, Juan C.

    2014-01-01

    SUMMARY Dietary changes may partly explain the high burden of asthma in industrialized nations. Experimental studies have motivated a significant number of observational studies of the relation between vitamins (A, C, D, and E) or nutrients acting as methyl donors (folate, vitamin B12, and choline) and asthma. Because observational studies are susceptible to several sources of bias, well-conducted randomized controlled trials (RCTs) remain the 揼old standard to determine whether a vitamin or nutrient has an effect on asthma. Evidence from observational studies and/or relatively few RCTs most strongly justify ongoing and future RCTs of: 1) vitamin D to prevent or treat asthma, 2) choline supplementation as adjuvant treatment for asthma, and 3) vitamin E to prevent the detrimental effects of air pollution in subjects with asthma. At this time, there is insufficient evidence to recommend supplementation with any vitamin or nutrient acting as a methyl donor to prevent or treat asthma. PMID:24461761

  20. Well-camouflaged Skin Graft Donor Sites in the Hand.

    PubMed

    Tan, Ruth E; Ying, Camelia Tang Qian; Sean, Lai Wei Hong; Lee, Ellen Y; Foo, Tun-Lin

    2015-12-01

    Three full-thickness skin graft sites within the hand where the donor sites are well blended with surrounding skin contour and crease are described. Two of these donor sites are located within the digit and these provide nonglaborous skin for small defects of the hand, whereas the third donor site along the palmar crease provides glaborous skin. These donor sites provide ease of access, minimal morbidity, and well-matched skin for resurfacing of small defects around the hand. The indications, contraindications, and technical points of each donor site are discussed. PMID:26296214

  1. Nitric oxide donors for cardiovascular implant applications.

    PubMed

    Naghavi, Noora; de Mel, Achala; Alavijeh, Omid Sadeghi; Cousins, Brian G; Seifalian, Alexander M

    2013-01-14

    In an era of increased cardiovascular disease burden in the ageing population, there is great demand for devices that come in to contact with the blood such as heart valves, stents, and bypass grafts that offer life saving treatments. Nitric oxide (NO) elution from healthy endothelial tissue that lines the vessels maintains haemostasis throughout the vasculature. Surgical devices that release NO are desirable treatment options and N-diazeniumdiolates and S-nitrosothiols are recognized as preferred donor molecules. There is a keen interest to investigate newer methods by which NO donors can be retained within biomaterials so that their release and kinetic profiles can be optimized. A range of polymeric scaffolds incorporating microparticles and nanomaterials are presenting solutions to current challenges, and have been investigated in a range of clinical applications. This review outlines the application of NO donors for cardiovascular therapy using biomaterials that release NO locally to prevent thrombosis and intimal hyperplasia (IH) and enhance endothelialization in the fabrication of next generation cardiovascular device technology. PMID:23136136

  2. Successful Treatment of Chronic Donor Site Pain

    PubMed Central

    Yanow, Jennifer H; Lorenzo, Luigi Di; Worosilo, Sharon C; Pappagallo, Marco

    2015-01-01

    Introduction: This is a case presentation of a 45-year-old male with chronic donor site pain following autologous iliac crest bone harvest successfully treated with superior cluneal nerve blockade. Donor site pain following autologous bone harvest is a common postoperative complication of lumbar fusion procedures that can cause significant morbidity and diminish quality of life, even in the context of an otherwise successful surgery. Dysfunction of the superior cluneal nerves is an etiology of this chronic pain. The patient抯 medical history, attempted treatments, and literature were reviewed. Case Presentation: A 45-year-old male with a six year history of severe pain over the right iliac crest following an otherwise successful lumbar laminectomy and fusion underwent two sets of superior cluneal nerve blocks, with sustained relief of more than 80% at seven months follow up. Conclusions: Donor site pain following autologous iliac crest bone harvest is a common surgical complication that is often resistant to conservative treatments such as physical therapy and oral medications. Blockade of the superior cluneal nerves is a safe and technically simple procedure that may result in long-term pain relief, obviating the need to consider more invasive options. PMID:26587399

  3. Serologic test-systems development: immunoassays for antibiotics. Progress report, May 16, 1980-September 30, 1981

    SciTech Connect

    Brake, R.; Hollstein, U.; Hindman, K.

    1983-04-01

    Progress on development of immunoassays for the antibiotics gentamicin, tetracycline, and tylosin is discussed. The development of the gentamicin assay was completed and the assay was transferred to the Beltsville Laboratories of the US Department of Agriculture (USDA) Food Safety and Quality Service (FSQS). The sensitivity (1 ppB) is 50-fold greater than we have previously reported, and is satisfactory for the current needs. At year's end, work was still in progress on both the tetracycline and tylosin assays. Tetracycline presents a more difficult problem of chemistry than we had anticipated. Immunoassay reagents synthesized by diazonium coupling were only weakly immunoreactive; analysis suggests that this coupling procedure damages the tetracycline structure. Several tetracycline derivatives that offer promising alternative coupling procedures were synthesized. Tylosin was successfully coupled to peroxidase. With this conjugate and antiserum, obtained from R. Mageau of the USDA, some immune-specific binding was demonstrated. Several problems with the tylosin assay remain to be resolved.

  4. Use of immunoassay testing and landfarming to remediate pesticide - contaminated soil at agrichemical businesses

    SciTech Connect

    Frank, J.F.

    1994-12-31

    The recent combination of two new technologies - immunoassay and landfarming - now offers an efficient, effective and economical option in the remediation of pesticide-contaminated soil. Participation in 28 landfarming projects in Illinois - 20 with pesticide-contaminated soil and eight with fuel-contaminated soil - has convinced the author that the combination offers the best method of remediation for most agrichemical sites. The procedures are currently used in Illinois as well as several other states and are being considered in even more. This paper describes the legislative and regulatory background for landfarming; defines immunoassay testing; defines landfarming: (1) contaminated site sampling and analysis; (2) education of participants; (3) selection of cooperator and host farm; (4) soil spreading considerations; a. rates, b. techniques; host site sampling and analysis; and factors affecting cost.

  5. Enzyme immunoassay and liquid chromatography-fluorescence detection for amikacin in raw milk.

    PubMed

    Chen, Yiqiang; Chen, Qian; He, Lidong; Shang, Bingru; Zhang, Liying

    2012-11-15

    An enzyme immunoassay and a liquid chromatography (LC) method for amikacin (AMK) in raw milk were developed in this study. Anti-AMK antibody was prepared by immunizing rabbits with AMK-BSA conjugate. The developed immunoassay exhibited an IC(50) value of 1.30 ng/mL and the spiked recoveries at 25-1000 ng/mL ranged from 69.8% to 93.3% with coefficients of variation (CVs) of 8.5-17.6%. For LC analysis, AMK was derivatized with 9-fluorenylmethyl chloroformate, which was followed by C8 column separation and fluorescence detection. By trichloroacetic acid extraction and MCX column purification, the recoveries at spiked concentrations of 500-5000 ng/mL were 80.7-91.3% with CVs less than 6.3%. The two methods can be selectively used for rapidly screening or quantitatively determining AMK in raw milk. PMID:22868103

  6. Chromatographic immunoassays: strategies and recent developments in the analysis of drugs and biological agents.

    PubMed

    Matsuda, Ryan; Rodriguez, Elliott; Suresh, Doddavenkatanna; Hage, David S

    2015-11-01

    A chromatographic immunoassay is a technique in which an antibody or antibody-related agent is used as part of a chromatographic system for the isolation or measurement of a specific target. Various binding agents, detection methods, supports and assay formats have been developed for this group of methods, and applications have been reported that range from drugs, hormones and herbicides to peptides, proteins and bacteria. This review discusses the general principles and applications of chromatographic immunoassays, with an emphasis being given to methods and formats that have been developed for the analysis of drugs and biological agents. The relative advantages or limitations of each format are discussed. Recent developments and research in this field, as well as possible future directions, are also considered. PMID:26571109

  7. Computer applications in the search for unrelated stem cell donors.

    PubMed

    M黮ler, Carlheinz R

    2002-08-01

    The majority of patients which are eligible for a blood stem cell transplantation from an allogeneic donor do not have a suitable related donor so that an efficient unrelated donor search is a prerequisite for this treatment. Currently, there are over 7 million volunteer donors in the files of 50 registries in the world and in most countries the majority of transplants are performed from a foreign donor. Evidently, computer and communication technology must play a crucial role in the complex donor search process on the national and international level. This article describes the structural elements of the donor search process and discusses major systematic and technical issues to be addressed in the development and evolution of the supporting telematic systems. The theoretical considerations are complemented by a concise overview over the current state of the art which is given by describing the scope, relevance, interconnection and technical background of three major national and international computer appliances: The German Marrow Donor Information System (GERMIS) and the European Marrow Donor Information System (EMDIS) are interoperable business-to-business e-commerce systems and Bone Marrow Donors World Wide (BMDW) is the basic international donor information desk on the web. PMID:12216954

  8. Transplanting Kidneys from Deceased Donors With Severe Acute Kidney Injury.

    PubMed

    Heilman, R L; Smith, M L; Kurian, S M; Huskey, J; Batra, R K; Chakkera, H A; Katariya, N N; Khamash, H; Moss, A; Salomon, D R; Reddy, K S

    2015-08-01

    Our aim was to determine outcomes with transplanting kidneys from deceased donors with acute kidney injury, defined as a donor with terminal serum creatinine ?2.0?mg/dL, or a donor requiring acute renal replacement therapy. We included all patients who received deceased donor kidney transplant from June 2004 to October 2013. There were 162 AKI donor transplant recipients (21% of deceased donor transplants): 139 in the standard criteria donor (SCD) and 23 in the expanded criteria donor (ECD) cohort. 71% of the AKI donors had stage 3 (severe AKI), based on acute kidney injury network (AKIN) staging. Protocol biopsies were done at 1, 4, and 12 months posttransplant. One and four month formalin-fixed paraffin embedded (FFPE) biopsies from 48 patients (24 AKI donors, 24 non-AKI) underwent global gene expression profiling using DNA microarrays (96 arrays). DGF was more common in the AKI group but eGFR, graft survival at 1 year and proportion with IF/TA>2 at 1 year were similar for the two groups. At 1 month, there were 898 differentially expressed genes in the AKI group (p-value <0.005; FDR <10%), but by 4 months there were no differences. Transplanting selected kidneys from deceased donors with AKI is safe and has excellent outcomes. PMID:25808278

  9. Overextended Criteria Donors: Experience of an Italian Transplantation Center.

    PubMed

    Nure, E; Lirosi, M C; Frongillo, F; Bianco, G; Silvestrini, N; Fiorillo, C; Sganga, G; Agnes, S

    2015-09-01

    The increasing gap between the number of patients who could benefit from liver transplantation and the number of available donors has fueled efforts to maximize the donor pool using marginal grafts that usually were discarded for transplantation. This study included data of all patients who received decreased donor liver grafts between January 2004 and January 2013 (n= 218) with the use of a prospectively collected database. Patients with acute liver failure, retransplantation, pediatric transplantation, and split liver transplantation were excluded. Donors were classified as standard donor (SD), extended criteria donor (ECD), and overextended criteria donor (OECD). The primary endpoints of the study were early allograft primary dysfunction (PDF), primary nonfunction (PNF), and patient survival (PS), whereas incidence of major postoperative complications was the secondary endpoint. In our series we demonstrated that OECD have similar outcome in terms of survival and incidence of complication after liver transplantation as ideal grafts. PMID:26361653

  10. Transplantation and differentiation of donor cells in the cloned pigs

    SciTech Connect

    Shimada, Arata; Tomii, Ryo; Kano, Koichiro; Nagashima, Hiroshi . E-mail: hnagas@isc.meiji.ac.jp

    2006-06-02

    The application of nuclear transfer technology is an interesting approach to investigate stem and progenitor cell transplantation therapy. If stem cells are used as a nuclear donor, donor cells can engraft into cloned animals without histocompatible problems. However, it is still uncertain whether donor cells can engraft to cloned animal and differentiate in vivo. To address this problem, we transplanted donor cells to dermal tissues of cloned pigs developed by using preadipocytes as donor cells. Preadipocytes are adipocytic progenitor which can differentiate to mature adipocytes in vitro. We showed that the donor preadipocytes were successfully transplanted into the cloned pigs without immune rejection and they differentiated into mature adipocytes in vivo 3 weeks after transplantation. In contrast, allogenic control preadipocytes, which can differentiate in vitro, did not differentiate in vivo. These results indicate that donor progenitor cells can differentiate in cloned animal.

  11. Magnetic Electrochemical Immunoassays with Quantum Dot Labels for Detection of Phosphorylated Acetylcholinesterase in Plasma

    SciTech Connect

    Wang, Hua; Wang, Jun; Timchalk, Charles; Lin, Yuehe

    2008-11-01

    A new magnetic electrochemical immunoassay has been developed as a tool for biomonitoring exposures to organophosphate (OP) compounds, e.g., insecticides and chemical nerve agents, by directly detecting organophosphorylated acetylcholinesterase (OP?AChE). This immunoassay uniquely incorporates highly efficient magnetic separation with ultrasensitive square wave voltammetry (SWV) analysis with quantum dots (QDs) as labels. A pair of antibodies was used to achieve the specific recognition of OP?AChE that was prepared with paraoxon as an OP model agent. Antiphosphoserine polyclonal antibodies were anchored on amorphous magnetic particles preferably chosen to capture OP?AChE from the sample matrixes by binding their phosphoserine moieties that were exposed through unfolding the protein adducts. This was validated by electrochemical examinations and enzyme-linked immunosorbent assays. Furthermore, antihuman AChE monoclonal antibodies were labeled with cadmium-source QDs to selectively recognize the captured OP?AChE, as characterized by transmission electron microscopy. The subsequent electrochemical SWV analysis of the cadmium component released by acid from the coupled QDs was conducted on disposable screen-printed electrodes. Experimental results indicated that the SWV-based immunoassays could yield a linear response over a broad concentration range of 0.3?300 ng/mL OP?AChE in human plasma with a detection limit of 0.15 ng/mL. Such a novel electrochemical immunoassay holds great promise as a simple, selective, sensitive, and field-deployable tool for the effective biomonitoring and diagnosis of potential exposures to nerve agents and pesticides.

  12. Improving of enzyme immunoassay for detection and quantification of the target molecules using silver nanoparticles

    NASA Astrophysics Data System (ADS)

    Syrvatka, Vasyl J.; Slyvchuk, Yurij I.; Rozgoni, Ivan I.; Gevkan, Ivan I.; Overchuk, Marta O.

    2014-02-01

    Modern routine enzyme immunoassays for detection and quantification of biomolecules have several disadvantages such as high cost, insufficient sensitivity, complexity and long-term execution. The surface plasmon resonance of silver nanoparticles gives reasons of creating new in the basis of simple, highly sensitive and low cost colorimetric assays that can be applied to the detection of small molecules, DNA, proteins and pollutants. The main aim of the study was the improving of enzyme immunoassay for detection and quantification of the target molecules using silver nanoparticles. For this purpose we developed method for synthesis of silver nanoparticles with hyaluronic acid and studied possibility of use these nanoparticles in direct determination of target molecules concentration (in particular proteins) and for improving of enzyme immunoassay. As model we used conventional enzyme immunoassays for determination of progesterone and estradiol concentration. We obtained the possibility to produce silver nanoparticles with hyaluronan homogeneous in size between 10 and 12 nm, soluble and stable in water during long term of storage using modified procedure of silver nanoparticles synthesis. New method allows to obtain silver nanoparticles with strong optical properties at the higher concentrations - 60-90 ?g/ml with the peak of absorbance at the wavelength 400 nm. Therefore surface plasmon resonance of silver nanoparticles with hyaluronan and ultraviolet-visible spectroscopy provide an opportunity for rapid determination of target molecules concentration (especial protein). We used silver nanoparticles as enzyme carriers and signal enhancers. Our preliminary data show that silver nanoparticles increased absorbance of samples that allows improving upper limit of determination of estradiol and progesterone concentration.

  13. Automation of the enzyme immunoassay for the serodiagnosis of infectious diseases in cattle

    SciTech Connect

    Seawright, G.L.; Sanders, W.M.; Bryson, M.

    1980-01-01

    The enzyme immunoassay (EIA) is a versatile and highly sensitive new tool that can be used to detect a wide variety of infectious diseases or toxic agents and other low molecular weight compound. The paper describes the present state of development of the EIA-modified Technicon Autoanalyzer II instrumentation, standardization of quality control criteria for the automated EIA and standardization of the diagnostic decision process. (ACR)

  14. Comparison of methaqualone excretion patterns using Abuscreen ONLINE and EMIT II immunoassays and GC/MS.

    PubMed

    Brenner, C; Hui, R; Passarelli, J; Wu, R; Brenneisen, R; Bracher, K; ElSohly, M A; Ghodoussi, V D; Salamone, S J

    1996-05-17

    A study was performed to compare the ONLINE and EMIT II immunoassays with gas chromatographic/mass spectrometric (GC/MS) analysis of methaqualone metabolites on urine using samples obtained from a clinical study. Urine was collected over a 72 h period from six healthy adults (4 male, 2 female) after oral dosing with 200 mg methaqualone (MTQ). Each urine sample was analyzed by ONLINE and EMIT II. The samples were then analyzed by GC/MS, hydrolyzed with beta-glucuronidase and again analyzed by GC/MS. Both immunoassays showed greater than 600 ng/ml concentrations of drug in each sample by the second void and remained highly positive for the rest of the 72 h. Unhydrolyzed samples analyzed by GC/MS showed both low concentrations of MTQ as well as its five major hydroxylated metabolites. The hydrolyzed samples analyzed by GC/MS showed high concentrations of the hydroxylated metabolites with the 2'-hydroxy and 3'-hydroxy metabolites being present at the highest concentrations, the 4'-hydroxy metabolite at a lower amount and the 6-hydroxy and 2-hydroxy metabolites at the lowest concentrations. The GC/MS data coupled with the antibody cross-reactivity data indicate that the major species in clinical samples that cross-react in both immunoassays are the conjugated forms of the hydroxylated metabolites of MTQ. Therefore when confirming by GC/MS after an immunoassay screen it would be prudent to confirm for the major hydroxylated metabolites as glucuronides of MTQ instead of the parent drug. PMID:8635771

  15. A nonenzymatic optical immunoassay strategy for detection of Salmonella infection based on blue silica nanoparticles.

    PubMed

    Sun, Qian; Zhao, Guangying; Dou, Wenchao

    2015-10-22

    A novel nonenzymatic optical immunoassay strategy was for the first time designed and utilized for sensitive detection of antibody to Salmonella pullorum and Salmonella gallinarum (S.爌ullorum and S.爂allinarum) in serum. The optical immunoassay strategy was based on blue silica nanoparticles (Blue-SiNps) and magnetic beads (MB). To construct such an optical immunoassay system, the Blue-SiNPs were first synthesized by inverse microemulsion method, characterized by SEM, Zeta potential and FTIR. Two nanostructures including Blue-SiNPs and MB were both functionalized with antibody against S.爌ullorum and S.爂allinarum (anti-PG) without using enzyme labeled antibody. Anti-PG functionalized blue silica nanoparticles (IgG-Blue-SiNps) were used as signal transduction labels, while anti-PG functionalized magnetic beads (IgG-MB) were selected to separate and enrich the final sandwich immune complexes. In the process of detecting negative serum, a sandwich immunocomplex is formed between the IgG-MB and IgG-Blue-SiNPs. With the separation of the immunocomplex using an external magnetic field, the final plaque displayed bright blue color. While in the detection of infected serum, IgG-MB and anti-PG formed sandwich immunocomplexes, IgG-Blue-SiNPs were unable to bind to the limited sites of the antigen, and a light brown plaque was displayed in the bottom of microplate well. Stable results were obtained with an incubation time of 60爉in at room temperature, and different colors corresponding to different results can be directly detected with naked eye. The reaction of IgG-Blue-SiNPs with S.爌ullorum was inhibited by 1:100 dilution of positive chicken serum. Such a simple immunoassay holds great potential as sensitive, selective and point-of-care (POC) tool for diagnosis of other biological molecules. PMID:26526916

  16. A competitive enzyme immunoassay for the quantitative detection of cocaine from banknotes and latent fingermarks.

    PubMed

    van der Heide, Susan; Garcia Calavia, Paula; Hardwick, Sheila; Hudson, Simon; Wolff, Kim; Russell, David A

    2015-05-01

    A sensitive and versatile competitive enzyme immunoassay (cEIA) has been developed for the quantitative detection of cocaine in complex forensic samples. Polyclonal anti-cocaine antibody was purified from serum and deposited onto microtiter plates. The concentration of the cocaine antibody adsorbed onto the plates, and the dilution of the cocaine-HRP hapten were both studied to achieve an optimised immunoassay. The method was successfully used to quantify cocaine in extracts taken from both paper currency and latent fingermarks. The limit of detection (LOD) of 0.162ngmL(-1) achieved with the assay compares favourably to that of conventional chromatography-mass spectroscopy techniques, with an appropriate sensitivity for the quantification of cocaine at the low concentrations present in some forensic samples. The cEIA was directly compared to LC-MS for the analysis of ten UK banknote samples. The results obtained from both techniques were statistically similar, suggesting that the immunoassay was unaffected by cross-reactivity with potentially interfering compounds. The cEIA was used also for the detection of cocaine in extracts from latent fingermarks. The results obtained were compared to the cocaine concentrations detected in oral fluid sampled from the same individual. Using the cEIA, we have shown, for the first time, that endogeneously excreted cocaine can be detected and quantified from a single latent fingermark. Additionally, it has been shown that the presence of cocaine, at similar concentrations, in more than one latent fingermark from the same individual can be linked with those concentrations found in oral fluid. These results show that detection of drugs in latent fingermarks could directly indicate whether an individual has consumed the drug. The specificity and feasibility of measuring low concentrations of cocaine in complex forensic samples demonstrate the effectiveness and robustness of the assay. The immunoassay presents a simple and cost-effective alternative to the current mass spectrometry based techniques for the quantitation of cocaine at forensically significant concentrations. PMID:25766738

  17. Detection of aerosolized biological agents by immunoassay followed by autonomous PCR confirmation

    SciTech Connect

    Dzenitis, J M; Hindson, B J; McBride, M T; Makarewicz, A J; Henderer, B D; Sathyam, U S; Smith, S M; Gutierrez, D M; Metz, T R; Venkateswaran, K S; Colston, B W; Farrow, S W

    2003-12-15

    An Autonomous Pathogen Detection System (APDS) unit is an automated, podium-sized system that monitors the air for all three biological threat agents (bacteria, viruses, and toxins). The system has been developed under the auspices of the U. S. Department of Energy and Department of Homeland Security by the University of California, Lawrence Livermore National Laboratory (LLNL) to protect people in critical or high-traffic facilities and at special events. The system performs continuous aerosol collection, sample preparation, and multiplexed biological tests using advanced immunoassays as the primary screen. Over ten agents are assayed at once, and results are reported hourly. R&D work this year focused on incorporating polymerase chain-reaction (PCR) techniques for detecting DNA as confirmation of immunoassay positives. The primary objective of the Dugway testing was to demonstrate the APDS with immunoassay identification and PCR confirmation of bacteria. A secondary objective was to demonstrate immunoassay identification of a protein toxoid (denatured toxin) aerosol release. A total of 12 agent trials were conducted over 14 days of testing, for a total of four work weeks at Dugway. Both testing objectives were achieved with multiple releases and clear identifications. The APDS was shown to be effective for identifying aerosolized Bacillus anthracis, Yersinia pestis, Bacillus globigii, and botulinum toxoid. The two areas for improvement were operational as opposed to hardware-related. The first was slowing the PCR thermal cycling to achieve stronger signals, which was demonstrated during the later phases of testing. The second area is to improve the parameters for autonomous PCR triggering; this is one of the focuses of the upcoming year's work.

  18. In situ deposition of Prussian blue on mesoporous carbon nanosphere for sensitive electrochemical immunoassay.

    PubMed

    Lai, Guosong; Zhang, Haili; Yu, Aimin; Ju, Huangxian

    2015-12-15

    A Prussian blue (PB) functionalized mesoporous carbon nanosphere (MCN) composite was prepared for loading signal antibody and high-content glucose oxidase (GOD) to obtain a new nanoprobe for sensitive electrochemical immunoassay. The MCN nanocarrier with an average diameter of 180 nm was synthesized by using mesoporous silica nanosphere as a hard template in combination with a hydrothermal carbonization method. This hydrophilic carbon nanomaterial provided an ideal platform for in situ deposition of high-content PB to form the MCN-PB nanocomposite. Based on the step-wise assembly of polyelectrolyte and gold nanoparticles (Au NPs) on the negative-charged nanocomposite, signal antibody and high-content GOD were loaded on this nanocarrier to obtain the nanoprobe. After a sandwich immunoreaction at an Au NPs-modified screen-printed carbon electrode based immunosensor, the nanoprobes were quantitatively captured on the electrode surface to produce sensitive electrochemical response with a PB-mediated GOD catalytic reaction for immunoassay. The high loading of PB and GOD on the nanoprobe greatly amplified the electrochemical signal, leading to the development of a new immunoassay method with high sensitivity. Using human immunoglobulin G as a model analyte, excellent analytical performance including a wide linear range from 0.01 to 100 ng/mL and a low detection limit down to 7.8 pg/mL was obtained. Additionally, the immunosensor showed high specificity, satisfactory stability and repeatability as well as acceptable reliability. The PB-mediated GOD electrochemical system well excluded the conventional interference from the dissolved oxygen. Thus this immunoassay method provides great potentials for practical applications. PMID:26201983

  19. Field-Portable Immunoassay Instruments and Reagents to Measure Chelators and Mobile Forms of Uranium

    SciTech Connect

    Blake, Diane A.

    2001-06-01

    Previous studies from our laboratory have demonstrated the feasibility of immunoassays for identification and quantification of specific metal ions. Our ultimate goal for this project is to (1) isolate and characterize antibodies that recognize the most mobile form of uranium, UO22+; (2) assemble, test, and validate a new field-portable immunosensor based on these antibodies; (3) prepare new monoclonal antibodies to the primary chelators (EDTA and DTPA) found in DOE wastes.

  20. Statistical approaches to developing a multiplex immunoassay for determining human exposure to environmental pathogens.

    PubMed

    Augustine, Swinburne A J; Simmons, Kaneatra J; Eason, Tarsha N; Griffin, Shannon M; Curioso, Clarissa L; Wymer, Larry J; Shay Fout, G; Grimm, Ann C; Oshima, Kevin H; Dufour, Al

    2015-10-01

    There are numerous pathogens that can be transmitted through water. Identifying and understanding the routes and magnitude of exposure or infection to these microbial contaminants are critical to assessing and mitigating risk. Conventional approaches of studying immunological responses to exposure or infection such as Enzyme-Linked Immunosorbent Assays (ELISAs) and other monoplex antibody-based immunoassays can be very costly, laborious, and consume large quantities of patient sample. A major limitation of these approaches is that they can only be used to measure one analyte at a time. Multiplex immunoassays provide the ability to study multiple pathogens simultaneously in microliter volumes of samples. However, there are several challenges that must be addressed when developing these multiplex immunoassays such as selection of specific antigens and antibodies, cross-reactivity, calibration, protein-reagent interferences, and the need for rigorous optimization of protein concentrations. In this study, a Design of Experiments (DOE) approach was used to optimize reagent concentrations for coupling selected antigens to Luminex xMAP microspheres for use in an indirect capture, multiplex immunoassay to detect human exposure or infection from pathogens that are potentially transmitted through water. Results from Helicobacter pylori, Campylobacter jejuni, Escherichia coli O157:H7, and Salmonella typhimurium singleplexes were used to determine the mean concentrations that would be applied to the multiplex assay. Cut-offs to differentiate between exposed and non-exposed individuals were determined using finite mixed modeling (FMM). The statistical approaches developed facilitated the detection of Immunoglobulin G (IgG) antibodies to H. pylori, C. jejuni, Toxoplasma gondii, hepatitis A virus, rotavirus and noroviruses (VA387 and Norwalk strains) in fifty-four diagnostically characterized plasma samples. Of the characterized samples, the detection rate was 87.5% for H. pylori, and 100% for T. gondii assays and 89% for HAV. Further, the optimized multiplex assay revealed exposure/infection to several other environmental pathogens previously uncharacterized in the samples. PMID:26070441

  1. Magnetic bead and gold nanoparticle probes based immunoassay for ?-casein detection in bovine milk samples.

    PubMed

    Li, Y S; Meng, X Y; Zhou, Y; Zhang, Y Y; Meng, X M; Yang, L; Hu, P; Lu, S Y; Ren, H L; Liu, Z S; Wang, X R

    2015-04-15

    In this work, a double-probe based immunoassay was developed for rapid and sensitive determination of ?-casein in bovine milk samples. In the method, magnetic beads (MBs), employed as supports for the immobilization of anti-?-casein polyclonal antibody (PAb), were used as the capture probe. Colloidal gold nanoparticles (AuNPs), employed as a bridge for loading anti-?-casein monoclonal antibody (McAb) and horseradish peroxidase (HRP), were used as the amplification probe. The presence of ?-casein causes the sandwich structures of MBs-PAb-?-casein-McAb-AuNPs through the interaction between ?-casein and the anti-?-casein antibodies. The HRP, used as an enzymatic-amplified tracer, can catalytically oxidize the substrate 3,3',5,5'-tetramethylbenzidine (TMB), generating optical signals that are proportional to the quantity of ?-casein. The linear range of the immunoassay was from 6.5 to 1520ngmL(-1). The limit of detection (LOD) was 4.8ngmL(-1) which was 700 times lower than that of MBs-antibody-HRP based immunoassay and 6-7 times lower than that from the microplate-antibody-HRP based assay. The recoveries of ?-casein from bovine milk samples were from 95.0% to 104.3% that had a good correlation coefficient (R(2)=0.9956) with those obtained by an official standard Kjeldahl method. For higher sensitivity, simple sample pretreatment and shorter time requirement of the antigen-antibody reaction, the developed immunoassay demonstrated the viability for detection of ?-casein in bovine milk samples. PMID:25522084

  2. Attitudes towards disclosure and relationship to donor offspring among a national cohort of identity-release oocyte and sperm donors

    PubMed Central

    Lampic, C.; Skoog Svanberg, A.; Sydsj, G.

    2014-01-01

    STUDY QUESTION What are oocyte donors and sperm donors' attitudes towards disclosure and relationship to donor offspring? SUMMARY ANSWER Oocyte and sperm donors in an identity-release donor programme support disclosure to donor offspring and have overall positive or neutral attitudes towards future contact with offspring. WHAT IS KNOWN ALREADY There is a global trend towards open-identity gamete donation with an increasing number of countries introducing legislation allowing only identifiable donors. While women and men who enrol in identity-release donor programmes accept that they may be contacted by donor offspring, there is limited knowledge of their attitudes towards disclosure to donor offspring and how they perceive their relationship to potential donor offspring. STUDY DESIGN, SIZE AND DURATION The present study is part of the 慡wedish study on gamete donation, a prospective cohort study including donors at all fertility clinics performing donation treatment in Sweden. During a 3-year period (20052008), donors were recruited consecutively and a total of 157 oocyte donors and 113 sperm donors (who did not donate to a specific 慿nown couple) were included prior to donation. Participants in the present study include 125 female (80%) and 80 male donors (71%) that completed two follow-up assessments. PARTICIPANTS/MATERIALS, SETTINGS AND METHODS Participants completed two postal questionnaires 2 months after donation and 14 months after donation. Attitudes towards disclosure to donor offspring were assessed with an established instrument. Perceptions of involvement with donor offspring and need for counselling was assessed with study-specific instruments. Statistical analyses were performed with non-parametric tests. MAIN RESULTS AND THE ROLE OF CHANCE A majority of oocyte and sperm donors supported disclosure to donor offspring (7191%) and had positive or neutral attitudes towards future contact with offspring (8087%). Sperm donors reported a higher level of involvement with potential donor offspring compared with oocyte donors (P = 0.005). Few donors reported a need for more counselling regarding the consequences of their donation. LIMITATIONS, REASONS FOR CAUTION While the multicentre study design strengthens external validity, attrition induced a risk of selection bias. In addition, the use of study-specific instruments that have not been psychometrically tested is a limitation. WIDER IMPLICATIONS OF THE FINDINGS The positive attitudes towards disclosure to offspring of female and male identity-release donors are in line with previous reports of anonymous and known donors. While our results on donors' general positive or neutral attitudes towards future contact with potential donor offspring are reassuring, a subset of donors with negative attitudes towards such contact warrants concern and suggests a need for counselling on long-term consequences of donating gametes. STUDY FUNDING The 慡wedish study on gamete donation was funded by the Swedish Research Council, the Swedish Council for Health, Working Life and Welfare, and the Regional Research Council in Uppsala-謗ebro. There are no conflicts of interest to declare. PMID:25030191

  3. Risk-adapted donor lymphocyte infusion based on chimerism and donor source in pediatric leukemia.

    PubMed

    Rujkijyanont, P; Morris, C; Kang, G; Gan, K; Hartford, C; Triplett, B; Dallas, M; Srinivasan, A; Shook, D; Pillai, A; Pui, C-H; Leung, W

    2013-01-01

    Donor lymphocyte infusion (DLI) is commonly used to treat leukemia relapse following stem cell transplantation. In florid relapse, however, the efficacy of DLI is limited with substantial risk of severe graft-versus-host disease (GvHD). Here, we develop a novel risk-adapted strategy characterized by pre-emptive DLI initiated at the time of mixed chimerism, a small starting dose based on donor source, dose-escalation guided by real-time chimerism monitoring and withholding of DLI immediately in patients achieving full donor chimerism. A total of 178 DLIs were given to 38 patients with mixed chimerism; thereafter, 33 patients (86.8%) had donor chimerism successfully increased, including 30 (78.9%) who had chimerism fully converted back to 100% donor. Cumulative incidence of relapse was significantly lower (P=0.00004) and overall survival higher (P=0.0003) in patients with chimerism fully corrected as compared with those of patients whose chimerism remained mixed. Only 13.2% of the patients developed acute grade III-IV GvHD with no associated mortality. In conclusion, the risk-adapted DLI strategy is useful in minimizing the risk of childhood leukemia relapse, GvHD and death. PMID:23995046

  4. Simulation of the kinetics of phosphorescence buildup of energy donor molecules in matrix-isolated donor-acceptor pairs

    NASA Astrophysics Data System (ADS)

    Zhdanova, N. V.; Deryabin, M. I.; Tishchenko, A. B.

    2015-10-01

    A mathematical model is proposed for describing the kinetics of accumulation of donor molecules in the triplet state for matrix-isolated donor-acceptor pairs. Using this model, computer simulation of the kinetics of accumulation of triplet excitations and phosphorescence buildup of the donor is performed. It is found that for certain relations between the rate constants of transitions for the donor and acceptor of the triplet excitation energy, the kinetics of phosphorescence buildup and the quantum yield of the donor phosphorescence depend on the excitation power.

  5. A Novel Colorimetric Immunoassay Utilizing the Peroxidase Mimicking Activity of Magnetic Nanoparticles

    PubMed Central

    Woo, Min-Ah; Kim, Moon Il; Jung, Jae Hwan; Park, Ki Soo; Seo, Tae Seok; Park, Hyun Gyu

    2013-01-01

    A simple colorimetric immunoassay system, based on the peroxidase mimicking activity of Fe3O4 magnetic nanoparticles (MNPs), has been developed to detect clinically important antigenic molecules. MNPs with ca. 10 nm in diameter were synthesized and conjugated with specific antibodies against target molecules, such as rotaviruses and breast cancer cells. Conjugation of the MNPs with antibodies (MNP-Abs) enabled specific recognition of the corresponding target antigenic molecules through the generation of color signals arising from the colorimetric reaction between the selected peroxidase substrate, 3,3?,5,5?-tetramethylbenzidine (TMB) and H2O2. Based on the MNP-promoted colorimetric reaction, the target molecules were detected and quantified by measuring absorbance intensities corresponding to the oxidized form of TMB. Owing to the higher stabilities and economic feasibilities of MNPs as compared to horseradish peroxidase (HRP), the new colorimetric system employing MNP-Abs has the potential of serving as a potent immunoassay that should substitute for conventional HRP-based immunoassays. The strategy employed to develop the new methodology has the potential of being extended to the construction of simple diagnostic systems for a variety of biomolecules related to human cancers and infectious diseases, particularly in the realm of point-of-care applications. PMID:23665902

  6. Development of Immunoassays Using Interferometric Real-Time Registration of Their Kinetics

    PubMed Central

    Orlov, A. V.; Burenin, A. G.; Shipunova, V. O.; Lizunova, A. A.; Gorshkov, B. G.; Nikitin, P. I.

    2014-01-01

    A method for effective development of solid-phase immunoassays on a glass surface and for optimization of related protocols by highly sensitive quantitative monitoring of each assay step has been proposed and experimentally implemented. The method is based on the spectral correlation interferometry (SCI) that allows real-time measuring of the thickness of a biomolecular layer bound to the recognition molecular receptors on the sensor chip surface. The method is realized with compact 3-channel SCI-biosensors that employ as the sensor chips standard cover glass slips without deposition of any additional films. Different schemes for antibody immobilization on a glass surface have been experimentally compared and optimized toward a higher sorption capacity of the sensor chips. Comparative characterization of the kinetics of each immunoassay stage has been implemented with the optimized protocols: i) covalent immobilization of antibody on an epoxylated surface and ii) biotinylated antibody sorption on a biotinylated surface via a high-affinity biotin-streptavidin bond. We have shown that magnetic nanoparticles employed as labels with model detection of cardiac troponin I further amplify the SCI signal, resulting in 100-fold improvement of the detection limit. The developed protocols can also be used with the alternative immunoassay platforms, including the label methods based on registration of only the final assay result, which is the quantity of bound labels. PMID:24772331

  7. Indirect competitive immunoassay for detection of vitamin B? in foods and pharmaceuticals.

    PubMed

    Wang, Peng; Yin, Yongmei; Eremin, Sergei A; Rybakov, Victor B; Zhang, Taichang; Xu, Zhihuan; Ren, Linlin; He, Xiaodan; Meng, Meng; Xi, Rimo

    2013-07-24

    An indirect immunoassay for the determination of vitamin B2 in food samples and vitamin tablets was developed. A carbodiimide-modified active ester method was used to synthesize the immunogen for vitamin B2. The coupling ratio of vitamin B2 to carrier protein in immunogen was 19.98:1. The titer of the polyclonal antibody was 1:64000, and the antibody showed high specificity in the presence of vitamin B2 photolytic products and other B group vitamins. The immunoassay showed detection limits (LODs) of 1.07 ng/mL in PBS, 24.6 ng/g in vitamin drink, and 0.50 mg/kg in milk powder. Recovery was 99.58-110.91% in milk powder and 70.20-100.5% in vitamin drink. Vitamin B2 samples were analyzed by high-pressure liquid chromatography (HPLC) and the immunoassay, and results showed good agreement. Finally, this method was applied to detect vitamin B2 in commercial milk powder and vitamin tablets, and the detected amount correlated well with the labeled amount. PMID:23855378

  8. Apoferritin Templated Synthesis of Metal Phosphate Nanoparticle Labels for Electrochemical Immunoassay

    SciTech Connect

    Liu, Guodong; Wu, Hong; Wang, Jun; Lin, Yuehe

    2006-08-29

    W have introduced template-synthesized metal phosphate nanoparticle labels for electrochemical immunoassay. Such use of an apoferritin template offers a simple and convenient route to prepare metallic nanoparticle labels for electrochemical immunoassays and avoid the complicated and time-consuming nanoparticle synthesis process (QD synthesis). Releasing metal ions from metal phosphate in an acetate buffer (pH 4.6) eliminates the harsh condition in the traditional metallic nanoparticle dissolution (e.g., strong acid dissolution of QDs and gold nanoparticles). This method is ultrasensitive and its DL is low to 77fM. The simultaneous detection of multiple protein targets is easily performed by using different metal phosphate nanoparticle labels (cadmium phosphate and lead phosphate). This approach can be extended to prepare multiple metal (such as zinc, lead, cadmium, copper, indium, gold, silver) phosphate nanoparticle labels or hybrid metal (bimetallic or trimetallic with predetermined ratios) phosphate nanoparticle labels for a multiplex electrochemical immunoassay. The new nanoparticle labels could be applicable to other electrochemical bioassays, such as DNA, and is thus expected to lead to wide applications for protein diagnostics and for bioanalysis in general.

  9. Method for Sorting and Pairwise Selection of Nanobodies for the Development of Highly Sensitive Sandwich Immunoassays.

    PubMed

    Rossotti, Mart韓 A; Pirez, Macarena; Gonzalez-Techera, Andres; Cui, Yongliang; Bever, Candace S; Lee, Kin S S; Morisseau, Christophe; Leizagoyen, Carmen; Gee, Shirley; Hammock, Bruce D; Gonz醠ez-Sapienza, Gualberto

    2015-12-01

    Single domain heavychain binders (nanobodies) obtained from camelid antibody libraries hold a great promise for immunoassay development. However, there is no simple method to select the most valuable nanobodies from the crowd of positive clones obtained after the initial screening. In this paper, we describe a novel nanobody-based platform that allows comparison of the reactivity of hundreds of clones with the labeled antigen, and identifies the best nanobody pairs for two-site immunoassay development. The output clones are biotinylated in vivo in 96-well culture blocks and then used to saturate the biotin binding capacity of avidin coated wells. This standardizes the amount of captured antibody allowing their sorting by ranking their reactivity with the labeled antigen. Using human soluble epoxide hydrolase (sEH) as a model antigen, we were able to classify 96 clones in four families and confirm this classification by sequencing. This provided a criterion to select a restricted panel of five capturing antibodies and to test each of them against the rest of the 96 clones. The method constitutes a powerful tool for epitope binning, and in our case allowed development of a sandwich ELISA for sEH with a detection limit of 63 pg/mL and four log dynamic range, which performed with excellent recovery in different tissue extracts. This strategy provides a systematic way to test nanobody pairwise combinations and would have a broad utility for the development of highly sensitive sandwich immunoassays. PMID:26544909

  10. Single-cell pathogen detection with a reverse-phase immunoassay on impedimetric transducers.

    PubMed

    de la Rica, Roberto; Baldi, Antonio; Fern醤dez-S醤chez, C閟ar; Matsui, Hiroshi

    2009-09-15

    The risk of infectious diseases has compelled some industries to establish a zero-tolerance standard for the presence of microorganisms in a given sample. Here, we address this issue with a novel reverse-phase immunoassay on impedimetric transducers for the specific detection of extremely low numbers of pathogens (less than 10 cells). After simply spotting the sample onto the electrodes, physisorbed analytes were targeted with urease-labeled antibodies, and the urease on the pathogens hydrolyzed urea to ionic species with a concomitant decrease of the resistivity of the solution. By this methodology, the limit of detection (LOD) based on the 3sigma criterion was 1 Escherichia coli cell with an assay time under 1 h. However, the precise number of cells present in highly diluted samples is uncertain, making it difficult to assess the final LOD of the sensor. We overcome this problem by using an atomic force microscope to deposit and image in situ the exact number cells on the transducer. After performing the immunoassay, a single E. coli cell was successfully detected without ambiguity in the number of cells even in the presence of a 10(4) excess of a competing microorganism, thus demonstrating the outstanding LOD and selectivity of the proposed reverse-phase immunoassay. PMID:19746997

  11. Blu-ray optomagnetic measurement based competitive immunoassay for Salmonella detection.

    PubMed

    Tian, Bo; Bejhed, Rebecca S; Svedlindh, Peter; Str鰉berg, Mattias

    2016-03-15

    A turn-on competitive immunoassay using a low-cost Blu-ray optomagnetic setup and two differently sized magnetic particles (micron-sized particles acting as capture particles and nano-sized particles acting as detection particles) is here presented. For Salmonella detection, a limit of detection of 810(4)CFU/mL is achieved within a total assay time of 3h. The combination of a competitive strategy and an optomagnetic setup not only enables a turn-on read-out format, but also results in a sensitivity limit about a factor of 20 times lower than of volumetric magnetic stray field detection device based immunoassays. The improvement of sensitivity is enabled by the formation of immuno-magnetic aggregates providing steric hindrance protecting the interior binding sites from interaction with the magnetic nanoparticle labels. The formation of immuno-magnetic aggregates is confirmed by fluorescence microscopy. The system exhibits no visible cross-reaction with other common pathogenic bacteria, even at concentrations as high as 10(7)CFU/mL. Furthermore, we present results when using the setup for a qualitative and homogeneous biplex immunoassay of Escherichia coli and Salmonella typhimurium. PMID:26386328

  12. A sensitive surface-enhanced Raman scattering enzyme-catalyzed immunoassay of respiratory syncytial virus.

    PubMed

    Zhan, Lei; Zhen, Shu Jun; Wan, Xiao Yan; Gao, Peng Fei; Huang, Cheng Zhi

    2016-02-01

    Respiratory viruses have become a major global health challenge which would benefit from advances in screening methods for early diagnosis. Respiratory syncytial virus (RSV) is one of the most important pathogen causing severe lower respiratory tract infections. Here we present a novel surface-enhanced Raman scattering (SERS) enzyme-catalyzed immunoassay of RSV by employing peroxidase substrate 3, 3'-5, 5'-tetramethylbenzidine (TMB) as Raman molecule. Horseradish peroxidase (HRP) attached to the detection antibody in a novel sandwich immunoassay catalyzes the oxidation of TMB by H2O2 to give a radical cation (TMB(+)), which could be easily adsorbed on the negatively charged surface of silver nanoparticles (AgNPs) through electrostatic interaction, inducing the aggregation of AgNPs and thus giving a strong SERS signal. A linear relationship was obtained between the Raman intensity and the amount of RSV in the range from 0.5 to 20pg/mL, and the minimum detectable concentration of this SERS-based enzyme immunoassay was 0.05pg/mL, which was 20 times lower than that found in the colorimetric method. PMID:26653454

  13. Laboratory analysis of remotely collected oral fluid specimens for opiates by immunoassay.

    PubMed

    Niedbala, R S; Kardos, K; Waga, J; Fritch, D; Yeager, L; Doddamane, S; Schoener, E

    2001-01-01

    The performance characteristics of a method for detecting opiates (morphine, codeine, heroin, and 6-acetylmorphine [6-AM]) in oral fluid specimens were examined and compared with methods for urine specimens. The oral fluid was easily obtained using a simple device that collects between 1 and 1.5 mL of fluid for laboratory analysis. Simultaneously collected specimens from 60 known opiate abusers from a drug-treatment center were first tested using an immunoassay cutoff of 10 ng/mL in oral fluids and 2,000 ng/mL in urine. Using a second aliquot, opiate confirmation in urine was performed by gas chromatography-mass spectrometry (GC-MS) and in oral fluids by GC-MS-MS. The combined immunoassay and GC-MS-MS procedures were completed with less than 250 pL of oral fluid. Opiates identified in oral fluid specimens from heroin users included morphine, codeine, heroin, and 6-AM. The immunoassay was tested for precision, stability, and the effects of potential cross-reactants. The results yielded 93.6% agreement between oral fluid and urine, suggesting that oral fluid may be a reliable matrix for opiate detection. PMID:11499883

  14. The effects of laser welding on heterogeneous immunoassay performance in a microfluidic cartridge

    PubMed Central

    M鋘tymaa, Anne; Halme, Jussi; V鋖imaa, Lasse; Kallio, Pasi

    2011-01-01

    Sealing of a microfluidic cartridge is a challenge, because the cartridge commonly contains heat-sensitive biomolecules that must also be protected from contamination. In addition, the objective is usually to obtain a sealing method suitable for mass production. Laser welding is a rapid technique that can be accomplished with low unit costs. Even though the technique has been widely adopted in industry, the literature on its use in microfluidic applications is not large. This paper is the first to report the effects of laser welding on the performance of the heterogeneous immunoassay in a polystyrene microfluidic cartridge in which biomolecules are immobilized into the reaction surface of the cartridge before sealing. The paper compares the immunoassay performance of microfluidic cartridges that are sealed either with an adhesive tape or by use of laser transmission welding. The model analyte used is thyroid stimulating hormone (TSH). The results show that the concentration curves in the laser-welded cartridges are very close to the curves in the taped cartridges. This indicates, first, that laser welding does not cause any significant reduction in immunoassay performance, and second, that the polystyrene cover does not have significant effect on the signal levels. Interestingly, the coefficients of variance between parallel samples were lower in the laser-welded cartridges than in the taped cartridges. PMID:22685505

  15. False-positive interferences of common urine drug screen immunoassays: a review.

    PubMed

    Saitman, Alec; Park, Hyung-Doo; Fitzgerald, Robert L

    2014-09-01

    Urine drug screen (UDS) immunoassays are a quick and inexpensive method for determining the presence of drugs of abuse. Many cross-reactivities exist with other analytes, potentially causing a false-positive result in an initial drug screen. Knowledge of these potential interferents is important in determining a course of action for patient care. We present an inclusive review of analytes causing false-positive interferences with drugs-of-abuse UDS immunoassays, which covers the literature from the year 2000 to present. English language articles were searched via the SciFinder platform with the strings 'false positive [drug] urine' yielding 173 articles. These articles were then carefully analyzed and condensed to 62 that included data on causes of false-positive results. The discussion is separated into six sections by drug class with a corresponding table of cross-reacting compounds for quick reference. False-positive results were described for amphetamines, opiates, benzodiazepines, cannabinoids, tricyclic antidepressants, phencyclidine, lysergic acid diethylamide and barbiturates. These false-positive results support the generally accepted practice that immunoassay positive results are considered presumptive until confirmed by a second independent chemical technique. PMID:24986836

  16. 3-D microarray and its microfabrication-free fluidic immunoassay device.

    PubMed

    Liu, Yingshuai; Zhang, Yuanyuan; Lu, Zhisong; Li, Chang Ming

    2015-08-19

    Conventional 2-D microarray is known to have high-throughput detection capability; however, the sensing spots density is significantly hindered by the spot-to-spot distance (gap) requirement for eliminating cross-talks between adjacent spots. Herein a new conceptual 3-D microarray device is proposed to significantly improve the spots density. To demonstrate advantages of the 3-D array, a microfabrication-free fluidic immunoassay device is further made by simply coupling an antibodies-arrayed glass cuboid into a circular glass tube. Rapid, sensitive and high-throughput flow-through immunoassays were accomplished with the 3-D array-based device for detection limits of 10-100 pg mL(-1) and wide dynamic range over 4-5 orders of magnitude in human serum with cancer biomarkers ?-fetoprotein (AFP) and carcinoembryonic antigen (CEA) as model targets, holding great promise for practical clinical applications. The 3-D microarray device not only significantly increases the density of sensing spots, but also greatly enhances the mass transport for rapid immunoassay when using in a flow-through device. PMID:26343442

  17. Microarray immunoassay for phenoxybenzoic acid using polymer-functionalized lanthanide oxide nanoparticles as fluorescent labels

    NASA Astrophysics Data System (ADS)

    Nichkova, Mikaela; Dosev, Dosi; Gee, Shirley J.; Hammock, Bruce D.; Kennedy, Ian M.

    2005-11-01

    Fluorescent properties and low production cost makes lanthanide oxide nanoparticles attractive labels in biochemistry. Nanoparticles with different fluorescent spectra were produced by doping of oxides such as Y IIO 3 and Gd IIO 3 with different lanthanide ions (Eu, Tb, Sm) giving the possibility for multicolor labeling. Protein microarrays have the potential to play a fundamental role in the miniaturization of biosensors, clinical immunological assays, and protein-protein interaction studies. Here we present the application of fluorescent lanthanide oxide nanoparticles as labels in microarray-based immunoassay for phenoxybenzoic acid (PBA), a generic biomarker of human exposure to the highly potent insecticides pyrethroids. A novel polymer-based protocol was developed for biochemical functionalization of the nanoparticles. Microarrays of antibodies were fabricated by microcontact printing in line patterns onto glass substrates and immunoassays were successfully performed using the corresponding functionalized nanoparticles. The applicability of the fluorophore nanoparticles as reporters for detection of antibody-antigen interactions has been demonstrated for phenoxybenzoic acid (PBA)/anti-PBA IgG. The sensitivity of the competitive fluorescent immunoassay for PBA was similar to that of the corresponding ELISA.

  18. Surface-enhanced Raman Immunoassay (SERIA): detection of Bacillus globigii in ground water

    NASA Astrophysics Data System (ADS)

    Guicheteau, Jason A.; Christesen, Steven D.

    2004-12-01

    This work presents the development of new methodologies centering on surfaces with immunologically induced affinities for biomaterials in aqueous systems. The immunologically active surfaces concentrate the biomaterials at the interface and therefore eliminate the need for preconcentration steps. This results in a highly sensitive and rapid immunoassay technique. The very strong localized of surface enhanced Raman scattering (SERS) that occurs at noble metal surfaces is combined with the unparalleled selectivity of immunoassays. Localization of the SERS signal eliminates the problem of washing and allows assays to be performed without treatment steps associated with removing excess agents. Previous work with small illicit drug molecules and large microorganisms clearly demonstrates trace detection of species in aqueous environments is possible. This paper discusses further work to detect Bacillus globigii by couping surface enhanced Raman scattering with immunoassays (SERIA) using citrate reduced silver nanoparticles. The spores of B. globigii are used to simulate the behavior of another bacterium that forms spores-the potential biological warfare agent, Bacillus anthracis, the causative agent of anthrax.

  19. Ready Access to Proquinazid Haptens via Cross-Coupling Chemistry for Antibody Generation and Immunoassay Development

    PubMed Central

    Esteve-Turrillas, Francesc A.; Mercader, Josep V.; Parra, Javier; Agull, Consuelo; Abad-Somovilla, Antonio; Abad-Fuentes, Antonio

    2015-01-01

    Bioconjugate preparation is a fundamental step for antibody generation and immunoassay development to small chemical compounds. For analytical targets holding in their structure an aryl halogen atom, cross-coupling reactions may be a simple and efficient way to obtain functionalized derivatives; thus offering great potential to elicit robust and selective immune responses after being coupled to immunogenic carrier proteins. However, substitution of the halogen atom by an aliphatic chain might eventually compromise the affinity and specificity of the resulting antibodies. In order to address this issue, proquinazid, a new-generation fungicide with outstanding performance, was chosen as model analyte. Two functionalized derivatives differing in spacer arm rigidity were synthesized by Sonogashira cross-coupling chemistry. These haptens were covalently coupled to bovine serum albumin and the resulting immunoconjugates were employed for rabbit vaccination. Antibodies were tested for proquinazid recognition by direct and indirect competitive immunoassay, and IC50 values in the low nanomolar range were found, thus demonstrating the suitability of this straightforward synthetic strategy for the generation of immunoreagents to compounds bearing an aryl halide. Following antibody characterization, competitive immunoassays were developed and employed to determine proquinazid residues in grape musts, and their analytical performance was satisfactorily validated by comparison with GC朚S. Besides having described the development of the first immunochemical method for proquinazid analysis, an efficient functionalization approach for analytes comprising aryl halides is reported. PMID:26214507

  20. Trace analysis of pollutants by use of honeybees, immunoassays, and chemiluminescence detection.

    PubMed

    Girotti, S; Ghini, S; Maiolini, E; Bolelli, L; Ferri, E N

    2013-01-01

    Specific and sensitive analysis to reveal and monitor the wide variety of chemical contaminants polluting all environment compartments, feed, and food is urgently required because of the increasing attention devoted to the environment and health protection. Our research group has been involved in monitoring the presence and distribution of agrochemicals by monitoring beehives distributed throughout the area studied. Honeybees have been used both as biosensors, because the pesticides affect their viability, and as "contaminant collectors" for all environmental pollutants. We focused our research on the development of analytical procedures able to reveal and quantify pesticides in different samples but with a special attention to the complex honeybee matrix. Specific extraction and purification procedures have been developed and some are still under optimization. The analytes of interest were determined by gas or liquid chromatographic methods and by compound-specific or group-specific immunoassays in the ELISA format, the analytical performance of which was improved by introducing luminescence detection. The range of chemiluminescent immunoassays developed was extended to include the determination of completely different pollutants, for example explosives, volatile organic compounds (including benzene, toluene, ethylbenzene, xylenes), and components of plastics, for example bisphenol A. An easier and portable format, a lateral flow immunoassay (LFIA) was added to the ELISA format to increase application flexibility in these assays. Aspects of the novelty, the specific characteristics, the analytical performance, and possible future development of the different chromatographic and immunological methods are described and discussed. PMID:23064670

  1. [Psychological profile of potential organ donors and non-organ donors].

    PubMed

    Blanca, Mar韆 J; Rando, Bel閚; Frutos, Miguel A; L髉ez-Montiel, Gema

    2007-08-01

    The aim of this research was to analyse the psychological profile of potential organ donors and potential non-organ donors, from a sample of people with qualifications lower than a Bachelor's degree. The variables examined were prosocial behaviour, the scales of the Constructive Thinking Inventory, and the dimensions of personality of the Big-Five Questionnaire. The results show that non-organ donors have a lower score in prosocial behaviour, are less efficient in their actions, with a tendency for less thought before acting, and they tend to be prejudiced. The results also reveal that this group has lower degrees of cooperation and empathy. They tend to be less reflective, less scrupulous, less willing to persevere in their actions, less interested in culture, and less open to new ideas and values. The above results are discussed, bearing in mind the utility of this knowledge to professionals dedicated to organ donation. PMID:17617983

  2. Thermoelectric Performance of Donor-Acceptor-Donor Conjugated Polymers Based on Benzothiadiazole Derivatives

    NASA Astrophysics Data System (ADS)

    Ming, Shouli; Zhen, Shijie; Lin, Kaiwen; Zhao, Li; Xu, Jingkun; Lu, Baoyang; Wang, Liangying; Xiong, Jinhua; Zhu, Zhengzhou

    2015-06-01

    Donor-acceptor-donor conjugated polymers are superior to other thermoelectric organic materials because it is much easier to modify their structure to reduce the bandgap between the conduction and valence bands, which is desirable for thermoelectric materials with high Seebeck coefficients. Despite this, studies of the thermoelectric performance of donor-acceptor-donor conjugated polymers are rare. In this study, four low-bandgap donor-acceptor-donor conjugated polymers, poly(4,7-bis(2,3-dihydrothieno[3,4- b][1,4] dioxin-5-yl)benzo[ c][1,2,5]thiadiazole) (PEBTE), poly(4,7-bis(2,3-dihydrothieno[3,4- b][1,4]dioxin-5-yl)benzo[ c][1,2,5]selenadiazole) (PEBSeE), poly (4,7-bis(2,3-dihydrothieno[3,4- b][1,4]dioxin-5-yl)-[1,2,5]thiadiazolo [3,4- c] pyridine) (PEPTE), and poly(4,7-bis(2,3-dihydrothieno[3,4- b][1,4]dioxin-5-yl)-[1,2,5]selenadiazolo[3,4- c]pyridine) (PEPSeE), were deposited by electrochemical polymerization of 4,7-bis(2,3-dihydrothieno[3,4-b][1,4]dioxin-5-yl)benzo[c][1,2,5]thiadiazole (EBTE), 4,7-bis(2,3-dihydro-thieno[3,4-b][1,4] dioxin-5-yl)benzo[c][1,2,5]selenadiazole (EBSeE), 4,7-bis(2,3-dihydrothieno [3,4-b][1,4]dioxin-5-yl)-[1,2,5]thiadiazolo[3,4-c] pyridine (EPTE) and 4,7-bis (2,3-dihydrothieno[3,4-b][1,4]dioxin-5-yl)-[1,2,5] selenadiazolo[3,4-c]pyridine (EPSeE), respectively and their thermoelectric performance was investi- gated. Compared with polyselenophenes, PEBTE and PEBSeE in pressed pellets had higher electrical conductivity (10-1-101 S cm-1) but lower Seebeck coefficient (14.0 ?V K-1) at room temperature. Future work may focus on treatment of these donor-acceptor-donor polymers to improve their electrical conductivity and Seebeck coefficient, and further investigation of their thermoelectric performance.

  3. Beneficial effects on the renal function of both recipients and donors in living donor kidney transplantation.

    PubMed

    Oh, C K; Yoon, S N; Lee, B M; Kim, J H; Kim, S J; Kim, H; Shin, G T

    2008-09-01

    In living donor kidney transplantation, initial function of the donor kidneys is split into the remnant kidney and the recipient's implanted kidney. We addressed the questions whether the donor's remnant kidney function increases or decreases after donation and whether the donor's donated kidney is greater or less than that of the recipient's implanted kidney after transplantation. We measured and calculated the functional ratio of each kidney using technetium-99m diethylenetriaminepentaacetic acid (99mTcDTPA) as well as serum creatinine (Scr; mg/dL) and creatinine clearance (Ccr; mL/min/1.73 m2) using 24-hour urines from 100 donors. The Ccr was also calculated using the Cockcroft-Gault formula (Ccr-CG; mL/min/1.73 m2). Within 7 days postnephrectomy, we measured Scr, Ccr, and Ccr-CG of the remnant kidney. For recipients, the Scr, Ccr, and Ccr-CG of the implanted kidney with 24-hour urine were obtained within 7 days posttransplantation. The average Scr, Ccr, and Ccr-CG of the donors were 0.85 +/- 0.17 mg/dL, 110.4 +/- 20.8 mL/min/1.73 m2, and 82.8 +/- 17.3 mL/min/1.73 m2, respectively. After donation, the Ccr and Ccr-CG of remnant kidney increased from 54.5 +/- 11.4 and 40.8 +/- 9.3 mL/min/1.73 m2 to 68.0 +/- 14.3 and 53.6 +/- 11.6 mL/min/1.73 m2, respectively. The recipient Ccr and Ccr-CG of donated kidney also increased from 55.9 +/- 11.8 and 42.0 +/- 9.9 mL/min/1.73 m2 to 78.4 +/- 18.0 and 53.4 +/- 16.4 mL/min/1.73 m2, respectively. Kidney transplantation from a living donor should be encouraged with total functional benefit for both donors and recipients. PMID:18790221

  4. [Robotical laparoscopic nephrectomy for living donors].

    PubMed

    Pavard, D; Decock, A; Bruy鑢e, F

    2015-11-01

    Robotic surgery has been developed since the 2000s. Urology is the discipline that received the widest of the evolution of robotic surgery especially due to prostatectomy. Meanwhile kidney failure develops and kidney transplantation is unfortunately hampered by the number of cadaveric grafts available. Kidney living donation takes its rise, not including the development of minimally invasive surgery in which robotics has its place. We describe the environment of this development as well as the technique and the contribution of the operating nurse in the removal of kidney with the robotic approach in living donors. PMID:26471213

  5. Q-FISH measurement of hepatocyte telomere lengths in donor liver and graft after pediatric living-donor liver transplantation: donor age affects telomere length sustainability.

    PubMed

    Kawano, Youichi; Ishikawa, Naoshi; Aida, Junko; Sanada, Yukihiro; Izumiyama-Shimomura, Naotaka; Nakamura, Ken-ichi; Poon, Steven S S; Matsumoto, Koshi; Mizuta, Koichi; Uchida, Eiji; Tajiri, Takashi; Kawarasaki, Hideo; Takubo, Kaiyo

    2014-01-01

    Along with the increasing need for living-donor liver transplantation (LDLT), the issue of organ shortage has become a serious problem. Therefore, the use of organs from elderly donors has been increasing. While the short-term results of LDLT have greatly improved, problems affecting the long-term outcome of transplant patients remain unsolved. Furthermore, since contradictory data have been reported with regard to the relationship between donor age and LT/LDLT outcome, the question of whether the use of elderly donors influences the long-term outcome of a graft after LT/LDLT remains unsettled. To address whether hepatocyte telomere length reflects the outcome of LDLT, we analyzed the telomere lengths of hepatocytes in informative biopsy samples from 12 paired donors and recipients (grafts) of pediatric LDLT more than 5 years after adult-to-child LDLT because of primary biliary atresia, using quantitative fluorescence in situ hybridization (Q-FISH). The telomere lengths in the paired samples showed a robust relationship between the donor and grafted hepatocytes (r?=?0.765, p?=?0.0038), demonstrating the feasibility of our Q-FISH method for cell-specific evaluation. While 8 pairs showed no significant difference between the telomere lengths for the donor and the recipient, the other 4 pairs showed significantly shorter telomeres in the recipient than in the donor. Multiple regression analysis revealed that the donors in the latter group were older than those in the former (p?=?0.001). Despite the small number of subjects, this pilot study indicates that donor age is a crucial factor affecting telomere length sustainability in hepatocytes after pediatric LDLT, and that the telomeres in grafted livers may be elongated somewhat longer when the grafts are immunologically well controlled. PMID:24727734

  6. A brown dwarf mass donor in an accreting binary.

    PubMed

    Littlefair, S P; Dhillon, V S; Marsh, T R; G鋘sicke, Boris T; Southworth, John; Watson, C A

    2006-12-01

    A long-standing and unverified prediction of binary star evolution theory is the existence of a population of white dwarfs accreting from substellar donor stars. Such systems ought to be common, but the difficulty of finding them, combined with the challenge of detecting the donor against the light from accretion, means that no donor star to date has a measured mass below the hydrogen burning limit. We applied a technique that allowed us to reliably measure the mass of the unseen donor star in eclipsing systems. We were able to identify a brown dwarf donor star, with a mass of 0.052 +/- 0.002 solar mass. The relatively high mass of the donor star for its orbital period suggests that current evolutionary models may underestimate the radii of brown dwarfs. PMID:17158322

  7. A brown dwarf mass donor in an accreting binary

    E-print Network

    S. P. Littlefair; V. S. Dhillon; T. R. Marsh; Boris T. Gaensicke; John Southworth; C. A. Watson

    2006-12-08

    A long standing and unverified prediction of binary star evolution theory is the existence of a population of white dwarfs accreting from sub-stellar donor stars. Such systems ought to be common, but the difficulty of finding them, combined with the challenge of detecting the donor against the light from accretion means that no donor star to date has a measured mass below the hydrogen burning limit. Here we apply a technique which allows us to reliably measure the mass of the unseen donor star in eclipsing systems. We are able to identify a brown dwarf donor star, with a mass of 0.052+/-0.002 Msun. The relatively high mass of the donor star for its orbital period suggests that current evolutionary models may underestimate the radii of brown dwarfs.

  8. Consequences of dietary methyl donor supplements: Is more always better?

    PubMed

    Shorter, Kimberly R; Felder, Michael R; Vrana, Paul B

    2015-07-01

    Epigenetic mechanisms are now recognized to play roles in disease etiology. Several diseases increasing in frequency are associated with altered DNA methylation. DNA methylation is accomplished through metabolism of methyl donors such as folate, vitamin B12, methionine, betaine (trimethylglycine), and choline. Increased intake of these compounds correlates with decreased neural tube defects, although this mechanism is not well understood. Consumption of these methyl donor pathway components has increased in recent years due to fortification of grains and high supplemental levels of these compounds (e.g. vitamins, energy drinks). Additionally, people with mutations in one of the enzymes that assists in the methyl donor pathway (5-MTHFR) are directed to consume higher amounts of methyl donors to compensate. Recent evidence suggests that high levels of methyl donor intake may also have detrimental effects. Individualized medicine may be necessary to determine the appropriate amounts of methyl donors to be consumed, particularly in women of child bearing age. PMID:25841986

  9. Non-heart-beating donors: an excellent choice to increase the donor pool.

    PubMed

    Nu馿z, J R; Del Rio, F; Lopez, E; Moreno, M A; Soria, A; Parra, D

    2005-11-01

    A specific program was adopted to obtain organs, for transplant purposes from people who die at home or in the street from sudden or unexpected death (type I non-heart-beating donors [NHBD] according to the Maastricht classification). The objective of our program was to increase the donor pool by obtaining organs from well-selected potential donors who die at home, work, or in the street and are maintained on advanced life support (ALS) until hospital arrival. The great number of people who die in a previously healthy situation constitute an excellent source of organs for transplant purposes. Our program includes pre- and in-hospital attendance. Prehospital attendance is based on application of cardiopulmonary resuscitation in situ and ALS until arrival at hospital. In hospital, specific preservation maneuvers must be performed and family assessment and judge permission obtained. In the last 15 years, we developed a kidney transplant program with better results than transplants performed with organs obtained from encephalic death donors (EDD). A specific NHBD subprogram for lung transplant was developed with excellent results as well. We are now improving the liver transplant program. NHBD are an important source of human tissues, including pancreas islets. It is clear that NHBD are a great source of organs and tissues for transplant, and that this kind of program must be established in all countries in which legal regulations allow it. PMID:16386494

  10. Benzochalcogenodiazole-based donor-acceptor-acceptor molecular donors for organic solar cells.

    PubMed

    Ting, Hao-Chun; Chen, Yi-Hong; Lin, Li-Yen; Chou, Shu-Hua; Liu, Yi-Hung; Lin, Hao-Wu; Wong, Ken-Tsung

    2014-02-01

    Four new molecules with a donor-acceptor-acceptor (D-A-A) configuration, in which 2,1,3-benzoxadiazole or 2,1,3-benzoselenodiazole were adopted as the central bridging acceptor, were synthesized as electron donors for small-molecule organic solar cells. In conjunction with two previously reported 2,1,3-benzothiadiazole-based compounds, the influences of the benzochalcogenodiazole acceptor unit and the ditolylarylamine donor moiety on the molecular structure, electrochemical behavior, and optical properties of the materials were investigated systematically to obtain a clear structure-property relationship. Vacuum-deposited hybrid planar mixed-heterojunction devices fabricated with the new donors and C70 as the acceptor showed power conversion efficiencies in the range of 2.9-4.3 % under 1 sun (100 mW cm(-2) ) AM 1.5 G simulated solar illumination. The current density-voltage characteristics of solar cells at various light intensities were measured, which revealed a high bimolecular recombination. PMID:24488678

  11. 75 FR 58400 - Donor Management Research: Improvements in Clinical Management of Deceased Organ Donors

    Federal Register 2010, 2011, 2012, 2013, 2014

    2010-09-24

    ...The Health Resources and Services Administration (HRSA), Division of Transplantation, is soliciting input, feedback, and suggestions from researchers and interested parties within the organ donation and transplant community regarding guidance for a possible grant or contract that focuses on improvements in clinical management of deceased organ donors. Given the continued imbalance between the......

  12. Mesoporous carbon-enriched palladium nanostructures with redox activity for enzyme-free electrochemical immunoassay of brevetoxin B.

    PubMed

    Lin, Youxiu; Zhou, Qian; Lin, Yuping; Lu, Minghua; Tang, Dianping

    2015-08-01

    A new signal amplification strategy based on mesoporous carbon-enriched palladium nanostructure (MSC-PdNS) was designed for enzyme-free electrochemical immunoassay of brevetoxin B (BTB) in marine toxins. The assay was carried out on a BTB-bovine serum albumin-functionalized electrode by using monoclonal mouse anti-BTB-labeling MSC-PdNS as the signal-transduction tag. A competitive-type assay protocol was successfully introduced to develop a high-efficiency enzyme-free immunoassay accompanying the doped palladium nanostructure into MSC-PdNS toward reduction of H2O2. Under the optimal conditions, the catalytic current decreased with the increment of BTB concentration in the range from 0.01 to 10 ng mL(-1) with a detection limit (LOD) of 5.0 pg mL(-1) BTB at the 3s(blank) criterion. The selectivity and precision were acceptable. In addition, the methodology was further validated for assaying spiked seafood samples, and consistent results between the electrochemical immunoassay and the referenced enzyme immunoassay were obtained. Importantly, the enzyme-free electrochemical immunoassay provides a promising approach for rapid screening of marine toxin because of its simplicity, low cost, sensitivity, specificity and without the need of sample pretreatment. PMID:26320787

  13. Immunoassay for Capsular Antigen of Bacillus anthracis Enables Rapid Diagnosis in a Rabbit Model of Inhalational Anthrax

    PubMed Central

    Gates-Hollingsworth, Marcellene A.; Perry, Mark R.; Chen, Hongjing; Needham, James; Houghton, Raymond L.; Raychaudhuri, Syamal; Hubbard, Mark A.; Kozel, Thomas R.

    2015-01-01

    Inhalational anthrax is a serious biothreat. Effective antibiotic treatment of inhalational anthrax requires early diagnosis; the further the disease has progressed, the less the likelihood for cure. Current means for diagnosis such as blood culture require several days to a result and require advanced laboratory infrastructure. An alternative approach to diagnosis is detection of a Bacillus anthracis antigen that is shed into blood and can be detected by rapid immunoassay. The goal of the study was to evaluate detection of poly-?-D-glutamic acid (PGA), the capsular antigen of B. anthracis, as a biomarker surrogate for blood culture in a rabbit model of inhalational anthrax. The mean time to a positive blood culture was 26 5.7 h (mean standard deviation), whereas the mean time to a positive ELISA was 22 4.2 h; P = 0.005 in comparison with blood culture. A lateral flow immunoassay was constructed for detection of PGA in plasma at concentrations of less than 1 ng PGA/ml. Use of the lateral flow immunoassay for detection of PGA in the rabbit model found that antigen was detected somewhat earlier than the earliest time point at which the blood culture became positive. The low cost, ease of use, and rapid time to result of the lateral flow immunoassay format make an immunoassay for PGA a viable surrogate for blood culture for detection of infection in individuals who have a likelihood of exposure to B. anthracis. PMID:25942409

  14. Counted Sb donors in Si quantum dots

    NASA Astrophysics Data System (ADS)

    Singh, Meenakshi; Pacheco, Jose; Bielejec, Edward; Perry, Daniel; Ten Eyck, Gregory; Bishop, Nathaniel; Wendt, Joel; Luhman, Dwight; Carroll, Malcolm; Lilly, Michael

    2015-03-01

    Deterministic control over the location and number of donors is critical for donor spin qubits in semiconductor based quantum computing. We have developed techniques using a focused ion beam and a diode detector integrated next to a silicon MOS single electron transistor to gain such control. With the diode detector operating in linear mode, the numbers of ions implanted have been counted and single ion implants have been detected. Poisson statistics in the number of ions implanted have been observed. Transport measurements performed on samples with counted number of implants have been performed and regular coulomb blockade and charge offsets observed. The capacitances to various gates are found to be in agreement with QCAD simulations for an electrostatically defined dot. This work was performed, in part, at the Center for Integrated Nanotechnologies, a U.S. DOE Office of Basic Energy Sciences user facility. The work was supported by Sandia National Laboratories Directed Research and Development Program. Sandia National Laboratories is a multi-program laboratory operated by Sandia Corporation, a Lockheed-Martin Company, for the U. S. Department of Energy under Contract No. DE-AC04-94AL85000.

  15. Lack of health insurance in living kidney donors.

    PubMed

    Casagrande, Lisette H; Collins, Sarah; Warren, Abigail T; Ommen, Elizabeth S

    2012-01-01

    Living donors are recommended to receive lifelong routine health maintenance after donation. There has been little examination of health insurance status among living donors, despite the fact that lack of health insurance is likely to impede donors' ability to obtain long-term healthcare post-donation. We performed a retrospective chart review for all living kidney donors at our institution between 2004 and 2008 to determine insurance status, demographic, socioeconomic, and basic health characteristics. Twenty-three percent of donors were uninsured at the time of donation. Odds of being uninsured were significantly lower in donors who were older than 40 yr of age or who had at least a college education, and significantly higher in donors who were non-white, non-English-speaking, or non-US citizens. Odds of being uninsured did not differ according to whether donors were obese, hypertensive, or smokers. On multivariate analysis, only non-white race, non-US citizenship, and education level less than a college degree were associated with lack of insurance. Lack of health insurance is more prevalent in living kidney donors than in the general US population. Its disproportionate impact on minorities, non-citizens, and the less well educated is greater than that in the general population. PMID:22050216

  16. Psychological strain in urgent indications for living donor liver transplantation.

    PubMed

    Erim, Yesim; Beckmann, Mingo; Kroencke, Sylvia; Valentin-Gamazo, Camino; Malago, Massimo; Broering, Dieter; Rogiers, Xavier; Frilling, Andrea; Broelsch, Christoph E; Schulz, Karl-Heinz

    2007-06-01

    The ethical soundness of living donor liver transplantation (LDLT) in urgent indications is still under discussion. The aim of the survey was to investigate the psychological distress of donors in cases of hepatocellular carcinoma (HCC) or acute liver failure (ALF). In a prospective multicenter study (n = 123), health-related quality of life (QOL), anxiety, and depression were measured. The psychological distress of donors was correlated to the degree of urgency of the recipients' indication, which was classified as nonurgent, HCC, or ALF. During the donor evaluation prior to LDLT, the donors with recipients for HCC and ALF demonstrated significantly reduced mental QOL in comparison to donors for a nonurgent indication and to the German normative sample. Compared to healthy controls, anxiety and depression were significantly increased in donors for ALF. Three months after the transplantation, scores for mental QOL as well as for anxiety and depression improved and were within the normal range for the whole group as well as for the ALF donors. In conclusion, the psychological burden was temporary in nature. Our findings can be considered as arguments for the current practice to address family members as donors in cases of HCC and ALF. PMID:17539009

  17. Implications of Donor Disseminated Intravascular Coagulation on Kidney Allograft Recipients

    PubMed Central

    Shafique, Shahzad; McCullagh, Johanna; Diederich, Dennis A.; Winklhofer, Franz T.; Wetmore, James B.

    2011-01-01

    Summary Background and objectives Disseminated intravascular coagulation (DIC) is common in deceased kidney donors and is considered a relative contraindication to donation. The significance of donor DIC on recipient kidney function is poorly understood. Additionally, the significance of thrombocytopenia in recipients of kidneys from DIC-positive donors is understudied. Design, setting, participants, & measurements In a retrospective cohort of 162 kidney transplants, the presence of DIC in donors, the occurrence of thrombocytopenia in recipients, and risk factors for delayed or slow graft function (DGF/SGF) were assessed. The effects of DIC donor status on DGF/SGF in the study sample as a whole, and of thrombocytopenia on DGF/SGF in recipients of DIC-positive kidneys specifically, were examined using multiple logistic regression. Results DIC donor status was not associated with occurrence of DGF/SGF, but thrombocytopenia was significantly associated with DIC-positive donor status (P = 0.008). Thrombocytopenia was independently associated with DGF/SGF only in the recipients of DIC-positive kidneys (P = 0.005). Patient and graft survival at 1 year were not affected by donor DIC status or by thrombocytopenia status. Conclusions Donor DIC was not associated with short-term suboptimal graft function, defined as DGF/SGF, nor with long-term patient or graft survival. However, thrombocytopenia appears to portend DGF/SGF in recipients of DIC-positive kidneys and may be a clinical sign on which the basis of therapeutic decisions could be undertaken. PMID:21372214

  18. AGU donors and supporting members play role at Fall Meeting

    NASA Astrophysics Data System (ADS)

    Berwager, Brent

    2012-02-01

    More than 1800 of AGU's top donors attended the 2011 Fall Meeting, and many of them sought refuge in the newly improved donor lounges. To quietly recognize supporting members, the lounges offered quiet space to relax between sessions, enjoy refreshments, meet with other donors, or work. Supporting members and AGU donors of all levels played a bigger role than ever at the Fall Meeting. Through the generous support of longtime members and AGU supporters Bruce Tsurutani and Olga Verkhoglyadova, the Space Weather and Nonlinear Waves and Processes Prize was announced at the meeting and will be awarded for the first time at the 2013 Fall Meeting.

  19. Survivability of immunoassay reagents exposed to the space radiation environment on board the ESA BIOPAN-6 platform as a prelude to performing immunoassays on Mars.

    PubMed

    Derveni, Mariliza; Allen, Marjorie; Sawakuchi, Gabriel O; Yukihara, Eduardo G; Richter, Lutz; Sims, Mark R; Cullen, David C

    2013-01-01

    The Life Marker Chip (LMC) instrument is an immunoassay-based sensor that will attempt to detect signatures of life in the subsurface of Mars. The molecular reagents at the core of the LMC have no heritage of interplanetary mission use; therefore, the design of such an instrument must take into account a number of risk factors, including the radiation environment that will be encountered during a mission to Mars. To study the effects of space radiation on immunoassay reagents, primarily antibodies, a space study was performed on the European Space Agency's 2007 BIOPAN-6 low-Earth orbit (LEO) space exposure platform to complement a set of ground-based radiation studies. Two antibodies were used in the study, which were lyophilized and packaged in the intended LMC format and loaded into a custom-made sample holder unit that was mounted on the BIOPAN-6 platform. The BIOPAN mission went into LEO for 12 days, after which all samples were recovered and the antibody binding performance was measured via enzyme-linked immunosorbent assays (ELISA). The factors expected to affect antibody performance were the physical conditions of a space mission and the exposure to space conditions, primarily the radiation environment in LEO. Both antibodies survived inactivation by these factors, as concluded from the comparison between the flight samples and a number of shipping and storage controls. This work, in combination with the ground-based radiation tests on representative LMC antibodies, has helped to reduce the risk of using antibodies in a planetary exploration mission context. PMID:23286207

  20. Increasing the Supply of Kidneys for Transplantation by Making Living Donors the Preferred Source of Donor Kidneys

    PubMed Central

    Testa, Giuliano; Siegler, Mark

    2014-01-01

    Abstract At the present time, increasing the use of living donors offers the best solution to the organ shortage problem. The clinical questions raised when the first living donor kidney transplant was performed, involving donor risk, informed consent, donor protection, and organ quality, have been largely answered. We strongly encourage a wider utilization of living donation and recommend that living donation, rather than deceased donation, become the first choice for kidney transplantation. We believe that it is ethically sound to have living kidney donation as the primary source for organs when the mortality and morbidity risks to the donor are known and kept extremely low, when the donor is properly informed and protected from coercion, and when accepted national and local guidelines for living donation are followed. PMID:25546677

  1. The donor-conceived child's "Right to Personal Identity": the public debate on donor anonymity in the United Kingdom.

    PubMed

    Turkmendag, Ilke

    2012-01-01

    On 1 April 2005, with the implementation of the Human Fertilisation and Embryology Authority (Disclosure of Donor Information) Regulations 2004, United Kingdom law was changed to allow children born through gamete donation to access details identifying the donor. Drawing on trends in adoption law, the decision to abolish donor anonymity was strongly influenced by a discourse that asserted the 慶hild's right to personal identity. Through examination of the donor anonymity debate in the public realm, while adopting a social constructionist approach, this article discusses how donor anonymity has been defined as a social problem that requires a regulative response. It focuses on the child's 憆ight to personal identity claims, and discusses the genetic essentialism behind these claims. By basing its assumptions on an adoption analogy, United Kingdom law ascribes a social meaning to the genetic relatedness between gamete donors and the offspring. PMID:22530247

  2. Simultaneous screening analysis of 3-methyl-quinoxaline-2-carboxylic acid and quinoxaline-2-carboxylic acid residues in edible animal tissues by a competitive indirect immunoassay

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Immunoassays contribute greatly to veterinary drug residue analysis and food safety, but there are no reported immunoassays on simultaneously detecting MQCA and QCA, the marker residues for carbadox and olaquindox. It is extremely difficult to produce broad-specificity antibodies that bind both res...

  3. Effect of donor and recipient variables on the long-term live-donor renal allograft survival in children

    PubMed Central

    Wafa, Ehab W.; Shokeir, Ahmed A.; Akl, Ahmed; Hassan, Nabil; Fouda, Mohamed A.; El Dahshan, Kalid; Ghoneim, Mohamed A.

    2011-01-01

    Objective We aimed to analyse donor and recipient predictors of graft survival in children who received live-donor renal grafts. Patients and methods The study comprised 273 children who received live-donor renal transplants at our center between March 1976 and October 2010. The follow-up ranged from 6爉onths to 25爕ears. Donor variables included donor age, gender, donor/recipient body weight ratio (DR BWR), ABO blood groups, human leukocyte antigen, and DR mismatching. Donor-specific problems, e.g., ischemia time during surgery and number of renal arteries, were included. Recipient variables included recipient age, sex, original kidney disease, ischemia time, acute tubular necrosis (ATN) after transplantation, immunosuppression, number of acute rejection episodes, re-transplantation, and development of hypertension. Results Independent variables with a sustained effect on the 5- and 10-year graft survival on multivariate analysis were: ATN after transplant, number of acute rejections, hypertension, and DR BWR. At the last follow-up, 185 patients (67.8%) had a functioning graft, while 82 (30.0%) had graft failure. Only six patients (0.02%) were lost to follow-up. Conclusion Donor and recipient variables that affect short- and long-term graft survival in children with a live-donor renal allograft are DR BWR, number of acute rejections, ATN and hypertension after transplant. Considering these variables provides a better outcome.

  4. Psychosocial impact of pediatric living-donor kidney and liver transplantation on recipients, donors, and the family: a systematic review.

    PubMed

    Thys, Kristof; Schwering, Karl-Leo; Siebelink, Marion; Dobbels, Fabienne; Borry, Pascal; Schotsmans, Paul; Aujoulat, Isabelle

    2015-03-01

    Living-donor kidney and liver transplantation intend to improve pediatric recipients' psychosocial well-being, but psychosocial impact in recipients strongly depends upon the impact on the donor and the quality of family relations. We systematically reviewed quantitative and qualitative studies addressing the psychosocial impact of pediatric living-donor kidney and liver transplantation in recipients, donors, and the family. In accordance with the PRISMA guidelines, we systematically searched the databases Medline, Web of Knowledge, Cinahl, Embase, ERIC, and Google Scholar. We identified 23 studies that satisfied our inclusion criteria. Recipients had improved coping skills and satisfactory peer relationships, but also reported anxiety and depressive symptoms, worried about the future, and had a negative body image. Similarly, donors experienced increased self-esteem, empowerment, and community awareness, but also complained of postoperative pain and a lack of emotional support. With respect to family impact, transplantation generated a special bond between the donor and the recipient, characterized by gratitude and admiration, but also raised new expectations concerning the recipient's lifestyle. As psychological problems in recipients were sometimes induced by feelings of guilt and indebtedness toward the donor, we recommend more research on how gift exchange dynamics function within donor-recipient relationships, enrolling donors and recipients within the same study. PMID:25363518

  5. A Novel Europium Chelate Coated Nanosphere for Time-Resolved Fluorescence Immunoassay.

    PubMed

    Shen, Yifeng; Xu, Shaohan; He, Donghua

    2015-01-01

    A novel europium ligand 2,2',2'',2'''-(4,7-diphenyl-1,10-phenanthroline-2,9-diyl) bis (methylene) bis (azanetriyl) tetra acetic acid (BC-EDTA) was synthesized and characterized. It shows an emission spectrum peak at 610 nm when it is excited at 360 nm, with a large Stock shift (250 nm). It is covalently coated on the surface of a bare silica nanosphere containi free amino groups, using 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide hydrochloride and N-Hydroxysuccinimide. We also observed an interesting phenomenon that when BC-EDTA is labeled with a silica nanosphere, the chelate shows different excitation spectrum peaks of about 295 nm. We speculate that the carboxyl has a significant influence on its excitation spectrum. The BC-EDTA/Eu3+coated nanosphere could be used as a fluorescent probe for time-resolved fluorescence immunoassay. We labeled the antibody with the fluorescent nanosphere to develop a nanosphere based hepatitis B surface antigen as a time-resolved fluorescence immunoassay reagent, which is very easy to operate and eliminates potential contamination of Eu3+ contained in the environment. The analytical and functional sensitivities are 0.0037 ?g/L and 0.08 ?g/L (S/N?2.0) respectively. The detection range is 0.08-166.67 ?g/L, which is much wider than that of ELISA (0.2-5 ?g/L). It is comparable to the commercial dissociation-enhanced lanthanide fluoro-immunoassay system (DELFIA) reagents (0.2-145 ?g/L). We propose that it can fulfill clinical applications. PMID:26056826

  6. Capillary nano-immunoassays: advancing quantitative proteomics analysis, biomarker assessment, and molecular diagnostics.

    PubMed

    Chen, Jin-Qiu; Wakefield, Lalage M; Goldstein, David J

    2015-01-01

    There is an emerging demand for the use of molecular profiling to facilitate biomarker identification and development, and to stratify patients for more efficient treatment decisions with reduced adverse effects. In the past decade, great strides have been made to advance genomic, transcriptomic and proteomic approaches to address these demands. While there has been much progress with these large scale approaches, profiling at the protein level still faces challenges due to limitations in clinical sample size, poor reproducibility, unreliable quantitation, and lack of assay robustness. A novel automated capillary nano-immunoassay (CNIA) technology has been developed. This technology offers precise and accurate measurement of proteins and their post-translational modifications using either charge-based or size-based separation formats. The system not only uses ultralow nanogram levels of protein but also allows multi-analyte analysis using a parallel single-analyte format for increased sensitivity and specificity. The high sensitivity and excellent reproducibility of this technology make it particularly powerful for analysis of clinical samples. Furthermore, the system can distinguish and detect specific protein post-translational modifications that conventional Western blot and other immunoassays cannot easily capture. This review will summarize and evaluate the latest progress to optimize the CNIA system for comprehensive, quantitative protein and signaling event characterization. It will also discuss how the technology has been successfully applied in both discovery research and clinical studies, for signaling pathway dissection, proteomic biomarker assessment, targeted treatment evaluation and quantitative proteomic analysis. Lastly, a comparison of this novel system with other conventional immuno-assay platforms is performed. PMID:26048678

  7. A Novel Europium Chelate Coated Nanosphere for Time-Resolved Fluorescence Immunoassay

    PubMed Central

    Shen, Yifeng; Xu, Shaohan; He, Donghua

    2015-01-01

    A novel europium ligand 2, 2, 2拻, 2拻-(4, 7-diphenyl-1, 10-phenanthroline-2, 9-diyl) bis (methylene) bis (azanetriyl) tetra acetic acid (BC-EDTA) was synthesized and characterized. It shows an emission spectrum peak at 610 nm when it is excited at 360 nm, with a large Stock shift (250 nm). It is covalently coated on the surface of a bare silica nanosphere containi free amino groups, using 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide hydrochloride and N-Hydroxysuccinimide. We also observed an interesting phenomenon that when BC-EDTA is labeled with a silica nanosphere, the chelate shows different excitation spectrum peaks of about 295 nm. We speculate that the carboxyl has a significant influence on its excitation spectrum. The BC-EDTA/Eu3+coated nanosphere could be used as a fluorescent probe for time-resolved fluorescence immunoassay. We labeled the antibody with the fluorescent nanosphere to develop a nanosphere based hepatitis B surface antigen as a time-resolved fluorescence immunoassay reagent, which is very easy to operate and eliminates potential contamination of Eu3+ contained in the environment. The analytical and functional sensitivities are 0.0037 ?g/L and 0.08 ?g/L (S/N?2.0) respectively. The detection range is 0.08-166.67 ?g/L, which is much wider than that of ELISA (0.2-5?g/L). It is comparable to the commercial dissociation-enhanced lanthanide fluoro-immunoassay system (DELFIA) reagents (0.2-145?g/L). We propose that it can fulfill clinical applications. PMID:26056826

  8. A rapid, noninvasive immunoassay for frataxin: Utility in assessment of Friedreich ataxia

    PubMed Central

    Deutsch, Eric C.; Santani, Avni B.; Perlman, Susan L.; Farmer, Jennifer M.; Stolle, Catherine A.; Marusich, Michael F.; Lynch, David R.

    2010-01-01

    Friedreich ataxia (FRDA) is an autosomal recessive neurodegenerative disorder caused by reduced amounts of the mitochondrial protein frataxin. Frataxin levels in research studies are typically measured via Western blot analysis from patient fibroblasts, lymphocytes, or muscle biopsies; none of these is ideal for rapid detection in large scale clinical studies. Recently, a rapid, non-invasive lateral-flow immunoassay was developed to accurately measure picogram levels of frataxin protein and shown to distinguish lymphoblastoid cells from FRDA carriers, patients and controls. We expanded the immunoassay to measure frataxin directly in buccal cells and whole blood from a large cohort of controls, known carriers and patients typical of a clinical trial population. The assay in buccal cells shared a similar degree of variability with previous studies conducted in lymphoblastoid cells (~10% coefficient of variation in controls). Significant differences in frataxin protein quantity were seen between the mean group values of controls, carriers, and patient buccal cells (100, 50.2, and 20.9% of control, respectively) and in protein extracted from whole blood (100, 75.3, and 32.2%, respectively), although there was some overlap between the groups. In addition, frataxin levels were inversely related to GAA repeat length and correlated directly with age of onset. Subjects with one expanded GAA repeat and an identified frataxin point mutation also carried frataxin levels in the disease range. Some patients displaying an FRDA phenotype but carrying only a single identifiable mutation had frataxin levels in the FRDA patient range. One patient from this group has a novel deletion that included exons 2 and 3 of the FXN gene based on multiplex ligation-dependent probe amplification (MLPA) analysis of the FXN gene. The lateral flow immunoassay may be a useful means to non-invasively assess frataxin levels repetitively with minimal discomfort in FRDA patients in specific situations such as clinical trials, and as a complementary diagnostic tool to aid in identification and characterization of atypical patients. PMID:20675166

  9. Preparation and quality control of silver nanoparticle-antibody conjugate for use in electrochemical immunoassays.

    PubMed

    Szymanski, Mateusz S; Porter, Robert A

    2013-01-31

    Metal nanoparticle-antibody conjugates are often used as optical or electrochemical markers in applications like immunohistochemistry, lateral flow tests, biosensors and immunoassays. In order to serve that role, an antibody needs to be immobilized on the surface of the nanoparticle. This is easily done, as proteins bind to gold and silver nanoparticles spontaneously. However, this immobilization process might result in nanoparticle aggregation or the loss of the bioactivity of the conjugated antibodies. In this work the optimization of antibody immobilization on silver colloid in order to obtain conjugates with the best possible activity is investigated. The parameters investigated were the type of immobilization buffer, its molarity and pH, the nanoparticle/antibody ratio and also blocking and washing protocols to reduce non-specific binding. The functionality of the obtained conjugates was tested with electrochemical immunoassay. It was found out that the optimum environment for immobilization of an anti-myoglobin antibody on silver nanoparticles was 0.2M boric acid pH 6.5 with 10 ?g of antibody loading per 1 mL of silver colloid. For an anti-troponin antibody it was 0.1M boric acid pH 7.5 also with 10 ?g/mL of antibody loading. The main problem for silver conjugation was the tendency of silver nanoparticles to aggregate during the immobilization process, but by choosing the optimum conditions the aggregation problem was completely removed. Here it is demonstrated that by using the conjugates prepared with an optimized protocol an increase in the sensitivity of the assay 10 times can be achieved. The electrochemical immunoassay described here can be used as a test for quality control of conjugates and for the estimation of batch-to-batch variability. PMID:23153725

  10. Sequential Injection/Electrochemical Immunoassay for Quantifying the Pesticide Metabolite 3, 5, 6-Trichloro-2-Pyridinol

    SciTech Connect

    Liu, Guodong; Riechers, Shawn L.; Timchalk, Chuck; Lin, Yuehe

    2005-12-04

    An automated and sensitive sequential injection electrochemical immunoassay was developed to monitor a potential insecticide biomarker, 3, 5, 6-trichloro-2-pyridinol. The current method involved a sequential injection analysis (SIA) system equipped with a thin-layer electrochemical flow cell and permanent magnet, which was used to fix 3,5,6-trichloro-2-pyridinol (TCP) antibody coated magnetic beads (TCP-Ab-MBs) in the reaction zone. After competitive immunoreactions among TCP-Ab-MBs, TCP analyte, and horseradish peroxidase (HRP) labeled TCP, a 3, 3?, 5, 5?-tetramethylbenzidine dihydrochloride and hydrogen peroxide (TMB-H2O2) substrate solution was injected to produce an electroactive enzymatic product. The activity of HRP tracers was monitored by a square wave voltammetric scanning electroactive enzymatic product in the thin-layer flow cell. The voltammetric characteristics of the substrate and the enzymatic product were investigated under batch conditions, and the parameters of the immunoassay were optimized in the SIA system. Under the optimal conditions, the system was used to measure as low as 6 ng L-1 (ppt) TCP, which is around 50-fold lower than the value indicated by the manufacturer of the TCP RaPID Assay? kit (0.25 ug/L, colorimetric detection). The performance of the developed immunoassay system was successfully evaluated on tap water and river water samples spiked with TCP. This technique could be readily used for detecting other environmental contaminants by developing specific antibodies against contaminants and is expected to open new opportunities for environmental and biological monitoring.

  11. CCQM-P58.1: Immunoassay Quantitation of Human Cardiac Troponin I.

    NASA Astrophysics Data System (ADS)

    Bunk, David; Noble, James; Knight, Alex E.; Wang, Lili; Klauenberg, Katy; Walzel, Monika; Elster, Clemens

    2015-01-01

    The CCQM study P58.1 assessed the equivalence of immunoassay measurements between participating NMIs. The aim of P58.1 was to demonstrate the equivalence of immunoassay measurements to determine the mass concentration of the clinically-relevant protein human cardiac troponin I (cTnI) present at low concentration relative to the protein concentration of the sample matrix. The measurement equivalence was assessed using traceability to a common certified reference material. To quantify cTnI, participants used a homogeneous sandwich-based immunoassay with an enzymatic amplification step. The antibody format consisted of a single capture and single detection antibody (referred to as 1 + 1), both were supplied to study participants. In the previous P58 study, ELISA measurement results were compared between laboratories which all used common ELISA reagents (including 96-well plates), samples, a standard for the production of calibrants, and a detailed ELISA protocol, which were supplied by a single laboratory. The P58.1 study only utilized common samples, a standard of the production of calibrants, and a set of monoclonal antibodies (mAbs). Because much of the experimental procedure for the P58 study was essentially standardized across participating labs, the study primarily highlighted between-laboratory differences in plate sampling designs and in plate reader response. As the participants in the P58.1 study had to produce most of their own analytical reagents and develop their own measurement procedure, the study provides a better evaluation of the equivalence of ELISA measurements between the participating laboratories. Main text. To reach the main text of this paper, click on Final Report The final report has been peer-reviewed and approved for publication by CCQM.

  12. Dual-Mode SERS-Fluorescence Immunoassay Using Graphene Quantum Dot Labeling on One-Dimensional Aligned Magnetoplasmonic Nanoparticles.

    PubMed

    Zou, Fengming; Zhou, Hongjian; Tan, Tran Van; Kim, Jeonghyo; Koh, Kwangnak; Lee, Jaebeom

    2015-06-10

    A novel dual-mode immunoassay based on surface-enhanced Raman scattering (SERS) and fluorescence was designed using graphene quantum dot (GQD) labels to detect a tuberculosis (TB) antigen, CFP-10, via a newly developed sensing platform of linearly aligned magnetoplasmonic (MagPlas) nanoparticles (NPs). The GQDs were excellent bilabeling materials for simultaneous Raman scattering and photoluminescence (PL). The one-dimensional (1D) alignment of MagPlas NPs simplified the immunoassay process and enabled fast, enhanced signal transduction. With a sandwich-type immunoassay using dual-mode nanoprobes, both SERS signals and fluorescence images were recognized in a highly sensitive and selective manner with a detection limit of 0.0511 pg mL(-1). PMID:26006156

  13. Sensitive monoclonal antibody-based immunoassays for kresoxim-methyl analysis in QuEChERS-based food extracts.

    PubMed

    Mercader, Josep V; L髉ez-Moreno, Rosario; Esteve-Turrillas, Francesc A; Agull, Consuelo; Abad-Somovilla, Antonio; Abad-Fuentes, Antonio

    2014-04-01

    Kresoxim-methyl is nowadays widely used to combat a diversity of common diseases affecting high-value crops. In this article, we report the development and characterization of two novel immunoassays for the analysis of this pioneer strobilurin fungicide, and for the first time, a validation study with food samples was performed. A direct and an indirect competitive immunoassay based on a new anti-kresoxim-methyl monoclonal antibody were developed for sensitive and specific chemical analysis. Optimized assays showed limits of detection of 0.1 ?g/L. Fruit and vegetable samples were extracted with acetonitrile by the QuEChERS procedure and analyzed by the developed immunoassays after a simple dilution in buffer, affording limits of quantification below US and European maximum residue limits. Immunochemical results of samples from kresoxim-methyl-sprayed strawberry fields demonstrated good statistical agreement with gas chromatography coupled to mass spectrometry as reference technique. PMID:24611510

  14. Silver nanocube-mediated sensitive immunoassay based on surface-enhanced Raman scattering assisted by etched silicon nanowire arrays.

    PubMed

    Jiang, Tao; Zhang, Li; Zhou, Jun

    2014-11-21

    An immunoassay based on surface enhanced Raman scattering (SERS) has been developed using immuno-silver nanocubes (NCs) and silicon nanowires with high sensitivity. The features of the samples were characterized by XRD profiles, absorption spectra, SEM, TEM, and AFM images, EDS analyses, and SERS spectra. It was found that the obtained silver NCs maintained higher SERS activity than silver nanospheres. After the silver NCs were modified by an antibody and employed in immunoassay with silicon nanowire arrays as the substrate, the antigen concentration-dependent SERS spectra and dose-response calibration curves were obtained. The detection limit of a prostate-specific antigen in this immunosystem was as low as 2 10(-14) g ml(-1), attributing to the significant electromagnetic coupling effect generated between the densely packed silver NCs and the SERS-active silicon nanowire arrays. It can be inferred that this proposed strategy will show great advantages in sensitive immunoassay and other biochemical examinations. PMID:25243249

  15. Platelet donation drives: a novel initiative to recruit platelet donors.

    PubMed

    Tendulkar, Anita; Shah, Sneha; Patil, Dipali; Tambe, Manisha

    2014-06-01

    The most important strategy to ensure a safe and an adequate supply of blood and blood products is motivation, recruitment, selection and retention of voluntary non remunerated blood donors. With a view of the increased platelet necessity in our oncology setup, the first platelet donation drive in the city and to the best of our knowledge, in India was conducted by our hospital in November 2009. The aim was to identify target groups and expand our donor database. It was also essential that the donor's contribution is acknowledged and appropriately felicitated. A campaign called "Save a Life" was initiated and publicized locally. A core team consisting of Transfusion Medicine specialists, clinicians and an NGO (nongovernment organization) was formed. The best suitable date and venue were finalized for the platelet camp. The audience was addressed and willing donors were registered as volunteer platelet donors with our institute. In a span of 40 months, 15 platelet camps were organized in colleges, social organizations, and corporate offices. A total of 1035 donors were registered out of which, 382 (37%) donated platelets in our hospital. 125/382 (33.2%) donated Single Donor Platelets (SDP) more than once. The largest number of platelet donations by a single camp donor was 24 times. Due to multiple donations from donors, the SDP number was enhanced considerably and lead to addition of 699 SDP units to our inventory. The annual indoor and camp voluntary platelet donor numbers increased from 142 in 2006 to 631 in 2012 due to platelet drives. All platelet donations were altruistic as no incentives were offered to the donors. Ready availability of platelets and planning SDP inventory as per patient blood group requirements had a positive impact on clinical services. PMID:24793425

  16. Laser Surface Treatment of Plastics for a Single-Channel Multiple Immunoassays Chip

    NASA Astrophysics Data System (ADS)

    Ooie, Toshihiko; Tanaka, Masato; Yamachoshi, Yuji; Nakahara, Tomonori; Abe, Kaori; Kataoka, Masatoshi

    A novel micro immunoassay chip for detecting biomarkers in blood plasma has been developed by using an injector-based antibody immobilization. Since multiple detectors were made in a single channel, the chip was able to detect various biomarkers simultaneously. UV Laser processing of the substrate was performed to enhance immobilization of the antibody. In P1CP, which is a biomarker for osteoporosis, this simple plastic chip requires only 1 micro liter per lane of blood plasma and 30 min of reaction time, one twentieth the sample volume required for the ELISA-kit and six times the through put.

  17. An enzyme immunoassay for rat growth hormone - Applications to the study of growth hormone variants

    NASA Technical Reports Server (NTRS)

    Farrington, Marianne A.; Hymer, W. C.

    1987-01-01

    A sensitive and specific competitive enzyme immunoassay for rat growth hormone (GH) is described and its use in the detection of GH variants is demonstrated. In the present assay, soluble GH and GH adsorbed to a solid-phase support compete for monkey anti-GH antibody binding sites. The immobilized antibody-GH complex is detected and quantified using goat antimonkey immunoglobin G covalently conjugated to horseradish peroxidase. It is noted that the assay can be performed in 27 hours and that sensitivities in the range of 0.19 to 25 ng can be obtained in the region of 10 to 90 percent binding.

  18. Analysis of drugs in human tissues by supercritical fluid extraction/immunoassay

    NASA Astrophysics Data System (ADS)

    Furton, Kenneth G.; Sabucedo, Alberta; Rein, Joseph; Hearn, W. L.

    1997-02-01

    A rapid, readily automated method has been developed for the quantitative analysis of phenobarbital from human liver tissues based on supercritical carbon dioxide extraction followed by fluorescence enzyme immunoassay. The method developed significantly reduces sample handling and utilizes the entire liver homogenate. The current method yields comparable recoveries and precision and does not require the use of an internal standard, although traditional GC/MS confirmation can still be performed on sample extracts. Additionally, the proposed method uses non-toxic, inexpensive carbon dioxide, thus eliminating the use of halogenated organic solvents.

  19. An ultra-sensitive microfluidic immunoassay using living radical polymerization and porous polymer monoliths.

    SciTech Connect

    Abhyankar, Vinay V.; Singh, Anup K.; Hatch, Anson V.

    2010-07-01

    We present a platform that combines patterned photopolymerized polymer monoliths with living radical polymerization (LRP) to develop a low cost microfluidic based immunoassay capable of sensitive (low to sub pM) and rapid (<30 minute) detection of protein in 100 {micro}L sample. The introduction of LRP functionality to the porous monolith allows one step grafting of functionalized affinity probes from the monolith surface while the composition of the hydrophilic graft chain reduces non-specific interactions and helps to significantly improve the limit of detection.

  20. Enzyme immunoassay versus latex agglutination cryptococcal antigen assays in adults with non-HIV-related cryptococcosis.

    PubMed

    Panackal, Anil A; Dekker, John P; Proschan, Michael; Beri, Andrea; Williamson, Peter R

    2014-12-01

    We compared paired enzyme immunoassay (EIA) and latex agglutination (LA) assay results with 185 blood and 164 cerebrospinal fluid (CSF) samples from 44 and 33 non-HIV cryptococcosis patients, respectively. The LA assay cutoff of 1:256 in the blood and 1:32 in the CSF was most highly predictive of a positive EIA result. The EIA missed 18.4% detected by the LA assay in the blood samples and 7.8% detected by the LA assay in the CSF samples. We note here the improved sensitivity of the LA assay over the EIA in non-HIV patients. PMID:25253799

  1. Immunoassay and Nb2 lymphoma bioassay prolactin levels and mammographic density in premenopausal and postmenopausal women the Nurses' Health Studies.

    PubMed

    Rice, Megan S; Tworoger, Shelley S; Bertrand, Kimberly A; Hankinson, Susan E; Rosner, Bernard A; Feeney, Yvonne B; Clevenger, Charles V; Tamimi, Rulla M

    2015-01-01

    Higher circulating prolactin levels have been associated with higher percent mammographic density among postmenopausal women in some, but not all studies. However, few studies have examined associations with dense area and non-dense breast area breast or considered associations with prolactin Nb2 lymphoma cell bioassay levels. We conducted a cross-sectional study among 1,124 premenopausal and 890 postmenopausal women who were controls in breast cancer case-control studies nested in the Nurses' Health Study (NHS) and NHSII. Participants provided blood samples in 1989-1990 (NHS) or 1996-1999 (NHSII) and mammograms were obtained from around the time of blood draw. Multivariable linear models were used to assess the associations between prolactin levels (measured by immunoassay or bioassay) with percent density, dense area, and non-dense area. Among 1,124 premenopausal women, percent density, dense area, and non-dense area were not associated with prolactin immunoassay levels in multivariable models (p trends = 0.10, 0.18, and 0.69, respectively). Among 890 postmenopausal women, those with prolactin immunoassay levels in the highest versus lowest quartile had modestly, though significantly, higher percent density (difference = 3.01 percentage points, 95 % CI 0.22, 5.80) as well as lower non-dense area (p trend = 0.02). Among women with both immunoassay and bioassay levels, there were no consistent differences in the associations with percent density between bioassay and immunoassay levels. Postmenopausal women with prolactin immunoassay levels in the highest quartile had significantly higher percent density as well as lower non-dense area compared to those in the lowest quartile. Future studies should examine the underlying biologic mechanisms, particularly for non-dense area. PMID:25503962

  2. Rare blood donors: a personal approach.

    PubMed

    Levene, C; Asher, O; Shinar, E; Yahalom, V

    2006-01-01

    The National Blood Group Reference Laboratory (NBGRL) in Israel was established in Jerusalem in 1971 and transferred to Magen David Adom (MDA), National Blood Services in 1995. This laboratory was the inspiration of the first author of this article for over 30 years. The realization of this vision was made possible by the cooperation of colleagues and laboratory workers in blood transfusion services throughout the country. The aim of the service was to provide diagnostic help in resolving immunohematologic problems found in the blood banks and clinics in Israel. In the beginning, only a part-time technician performed the work and testing was done using very limited reagents. The service was expanded by personal visits to all of the 22 blood banks in Israel to explain the aim of this new service and to educate them about the importance of resolving each and every case. One major issue was the cost involved in referring problems but it was decided at the outset that these would be covered by the government to ensure that a workup would be performed for all referred cases. The expansion of the service could not have been achieved without the help of the SCARF program. This voluntary service enabled us to identify the first rare donors in Israel, resolve complex cases, and find compatible blood for our patients. To illustrate the importance of the NBGRL in Israel and the rapid resolution of cases referred, several individual stories are described. The purpose of this review is to show the importance of the NBGRL in identifying rare blood groups and in providing and coordinating services and the importance of keeping in close contact with the rare donors to encourage and promote their donations, which may save lives. PMID:16813467

  3. [Skin allograft, from the donor to the burn patient].

    PubMed

    Gaucher, Sonia; Marchix, Thierry; Jarraya, Mohamed

    2012-01-01

    Severely burned patients are treated through the use of donor skin or skin allografts. It is a vital treatment. From the donor to the recipient, the process involves numerous procedures which are performed in accordance with ethical and medical safety rules. PMID:22916399

  4. Digging up Classroom Dollars on DonorsChoose

    ERIC Educational Resources Information Center

    Curriculum Review, 2006

    2006-01-01

    Back in 2000, Charles Best was teaching at Wings Academy, an alternative high school in the Bronx, when he got the idea for a Web site where teachers could solicit donations for class projects. With help from his students, DonorsChoose.org soon was born. Last year, the site won Amazon.com's Nonprofit Innovation Award. So far, DonorsChoose has

  5. Optical Control of Donor Spin Qubits in Silicon

    E-print Network

    M. J. Gullans; J. M. Taylor

    2015-10-26

    We show how to achieve optical, spin-selective transitions from the ground state to excited orbital states of group-V donors (P, As, Sb, Bi) in silicon. We consider two approaches based on either resonant, far-infrared (IR) transitions of the neutral donor or resonant, near-IR excitonic transitions. For far-IR light, we calculate the dipole matrix elements between the valley-orbit and spin-orbit split states for all the goup-V donors using effective mass theory. We then calculate the maximum rate and amount of electron-nuclear spin-polarization achievable through optical pumping with circularly polarized light. We find this approach is most promising for Bi donors due to their large spin-orbit and valley-orbit interactions. Using near-IR light, spin-selective excitation is possible for all the donors by driving a two-photon $\\Lambda$-transition from the ground state to higher orbitals with even parity. We show that externally applied electric fields or strain allow similar, spin-selective $\\Lambda$-transition to odd-parity excited states. We anticipate these results will be useful for future spectroscopic investigations of donors, quantum control and state preparation of donor spin qubits, and for developing a coherent interface between donor spin qubits and single photons.

  6. Optical control of donor spin qubits in silicon

    NASA Astrophysics Data System (ADS)

    Gullans, M. J.; Taylor, J. M.

    2015-11-01

    We show how to achieve optical, spin-selective transitions from the ground state to excited orbital states of group-V donors (P, As, Sb, and Bi) in silicon. We consider two approaches based on either resonant, far-infrared (IR) transitions of the neutral donor or resonant, near-IR excitonic transitions. For far-IR light, we calculate the dipole matrix elements between the valley-orbit and spin-orbit split states for all the group-V donors using effective mass theory. We then calculate the maximum rate and amount of electron-nuclear spin-polarization achievable through optical pumping with circularly polarized light. We find this approach is most promising for Bi donors due to their large spin-orbit and valley-orbit interactions. Using near-IR light, spin-selective excitation is possible for all the donors by driving a two-photon ? transition from the ground state to higher orbitals with even parity. We show that externally applied electric fields or strain allow similar, spin-selective ? transition to odd-parity excited states. We anticipate these results will be useful for future spectroscopic investigations of donors, quantum control and state preparation of donor spin qubits, and for developing a coherent interface between donor spin qubits and single photons.

  7. Lung procurement for transplantation: new criteria for lung donor selection.

    PubMed

    Moretti, M P; Betto, C; Gambacorta, M; Vesconi, S; Scalamogna, M; Benazzi, E; Ravini, M

    2010-05-01

    In Italy, like everywhere in the world, the organ shortage for transplantation is a real problem. It is well known that lung donors (LD) are particularly difficult to procure and that management of the organ do not care during the diagnosis of cerebral death represents a difficult challenge. In this context, the salvage of the so-called "marginal donors" may increase the pool of donors, favoring organ retrieval. To increase lung procurement, the intensivist must recognize "marginal donors," optimizing organ selection and function. The aim of our study was to review LD procured in 2008, as identified by the unrestricted criteria, of the Nord Italian Transplant program Center (NITp). Particularly, the age and habits of donors and the presence of a parenchyma contusion were not sufficient per se to exclude donation. We revisited lung ventilation and monitoring modalities during cerebral death before retrieval. In 2008, the application of enlarged criteria for LD enabled us to collect 21 LD, namely 33% of all cerebral deaths, versus 13% in 2007. Seeking to maintain good gas exchange and lung function, we implemented a safe ventilation program avoided high peak pressures, and fluid therapy properly guided by the cardiac index and extravascular lung water index monitoring. Specific actions to improve LD procurement may help cope with the organ-donor shortage. Although our series was small, our results were encouraging; they underline the necessity to continuously review donor criteria and care, allowing good donor/recipient matching. PMID:20534222

  8. Understanding Philanthropic Motivations of Northeast State Community College Donors

    ERIC Educational Resources Information Center

    Cook, Heather J.

    2012-01-01

    At Northeast State Community College (NeSCC) nearly 70% of students need some form of financial aid to attend. State support is flattening or decreasing and the gap is filled by private donors' support (Northeast State Community College, 2011). Hundreds of donors have made significant contributions to aid in the education of those in the

  9. Games Universities Play: And How Donors Can Avoid Them

    ERIC Educational Resources Information Center

    Wooster, Martin Morse

    2011-01-01

    What responsibilities do universities have to donors, many of whom are alumni? Presumably, one responsibility is to respect their wishes when they provide gifts with specific purposes. Yet Martin Morse Wooster shows in this report that universities often neglect the wishes of contributors. "Games Universities Play: And How Donors Can Avoid Them"

  10. Multivalley effective mass theory simulation of donors in silicon

    E-print Network

    John King Gamble; N. Tobias Jacobson; Erik Nielsen; Andrew D. Baczewski; Jonathan E. Moussa; In鑣 Monta駉; Richard P. Muller

    2015-11-02

    Last year, Salfi et al. made the first direct measurements of a donor wave function and found extremely good theoretical agreement with atomistic tight-binding [Salfi et al., Nat. Mater. 13, 605 (2014)]. Here, we show that multi-valley effective mass theory, applied properly, does achieve close agreement with tight-binding and hence gives reliable predictions. To demonstrate this, we variationally solve the coupled six-valley Shindo-Nara equations, including silicon's full Bloch functions. Surprisingly, we find that including the full Bloch functions necessitates a tetrahedral, rather than spherical, donor central cell correction to accurately reproduce the experimental energy spectrum of a phosphorus impurity in silicon. We cross-validate this method against atomistic tight-binding calculations, showing that the two theories agree well for the calculation of donor-donor tunnel coupling. Further, we benchmark our results by performing a statistical uncertainty analysis, confirming that derived quantities such as the wave function profile and tunnel couplings are robust with respect to variational energy fluctuations. Finally, we apply this method to exhaustively enumerate the tunnel coupling for all donor-donor configurations within a large search volume, demonstrating conclusively that the tunnel coupling has no spatially stable regions. Though this instability is problematic for reliably coupling donor pairs for two-qubit operations, we identify specific target locations where donor qubits can be placed with scanning tunneling microscopy technology to achieve reliably large tunnel couplings.

  11. Multivalley effective mass theory simulation of donors in silicon

    NASA Astrophysics Data System (ADS)

    Gamble, John King; Jacobson, N. Tobias; Nielsen, Erik; Baczewski, Andrew D.; Moussa, Jonathan E.; Monta駉, In鑣; Muller, Richard P.

    2015-06-01

    Last year, Salfi et al. made the first direct measurements of a donor wave function and found extremely good theoretical agreement with atomistic tight-binding theory results [Salfi et al., Nat. Mater. 13, 605 (2014), 10.1038/nmat3941]. Here, we show that multivalley effective mass theory, applied properly, does achieve close agreement with tight-binding results and hence gives reliable predictions. To demonstrate this, we variationally solve the coupled six-valley Shindo-Nara equations, including silicon's full Bloch functions. Surprisingly, we find that including the full Bloch functions necessitates a tetrahedral, rather than spherical, donor central cell correction to accurately reproduce the experimental energy spectrum of a phosphorus impurity in silicon. We cross-validate this method against atomistic tight-binding calculations, showing that the two theories agree well for the calculation of donor-donor tunnel coupling. Further, we benchmark our results by performing a statistical uncertainty analysis, confirming that derived quantities such as the wave function profile and tunnel couplings are robust with respect to variational energy fluctuations. Finally, we apply this method to exhaustively enumerate the tunnel coupling for all donor-donor configurations within a large search volume, demonstrating conclusively that the tunnel coupling has no spatially stable regions. Although this instability is problematic for reliably coupling donor pairs for two-qubit operations, we identify specific target locations where donor qubits can be placed with scanning tunneling microscopy technology to achieve reliably large tunnel couplings.

  12. 21 CFR 640.66 - Immunization of donors.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... 21 Food and Drugs 7 2010-04-01 2010-04-01 false Immunization of donors. 640.66 Section 640.66 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) BIOLOGICS ADDITIONAL STANDARDS FOR HUMAN BLOOD AND BLOOD PRODUCTS Source Plasma 640.66 Immunization of donors....

  13. 21 CFR 640.31 - Suitability of donors.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... 21 Food and Drugs 7 2010-04-01 2010-04-01 false Suitability of donors. 640.31 Section 640.31 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) BIOLOGICS ADDITIONAL STANDARDS FOR HUMAN BLOOD AND BLOOD PRODUCTS Plasma 640.31 Suitability of donors. (a)...

  14. 21 CFR 640.31 - Suitability of donors.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... 21 Food and Drugs 7 2011-04-01 2010-04-01 true Suitability of donors. 640.31 Section 640.31 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) BIOLOGICS ADDITIONAL STANDARDS FOR HUMAN BLOOD AND BLOOD PRODUCTS Plasma 640.31 Suitability of donors. (a)...

  15. 21 CFR 640.66 - Immunization of donors.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... 21 Food and Drugs 7 2011-04-01 2010-04-01 true Immunization of donors. 640.66 Section 640.66 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) BIOLOGICS ADDITIONAL STANDARDS FOR HUMAN BLOOD AND BLOOD PRODUCTS Source Plasma 640.66 Immunization of donors....

  16. 21 CFR 640.31 - Suitability of donors.

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ... 21 Food and Drugs 7 2012-04-01 2012-04-01 false Suitability of donors. 640.31 Section 640.31 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) BIOLOGICS ADDITIONAL STANDARDS FOR HUMAN BLOOD AND BLOOD PRODUCTS Plasma 640.31 Suitability of donors. (a)...

  17. 21 CFR 640.66 - Immunization of donors.

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ... 21 Food and Drugs 7 2012-04-01 2012-04-01 false Immunization of donors. 640.66 Section 640.66 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) BIOLOGICS ADDITIONAL STANDARDS FOR HUMAN BLOOD AND BLOOD PRODUCTS Source Plasma 640.66 Immunization of donors....

  18. 21 CFR 640.66 - Immunization of donors.

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ... 21 Food and Drugs 7 2013-04-01 2013-04-01 false Immunization of donors. 640.66 Section 640.66 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) BIOLOGICS ADDITIONAL STANDARDS FOR HUMAN BLOOD AND BLOOD PRODUCTS Source Plasma 640.66 Immunization of donors....

  19. 21 CFR 640.31 - Suitability of donors.

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ... 21 Food and Drugs 7 2013-04-01 2013-04-01 false Suitability of donors. 640.31 Section 640.31 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) BIOLOGICS ADDITIONAL STANDARDS FOR HUMAN BLOOD AND BLOOD PRODUCTS Plasma 640.31 Suitability of donors. (a)...

  20. 21 CFR 640.66 - Immunization of donors.

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ... 21 Food and Drugs 7 2014-04-01 2014-04-01 false Immunization of donors. 640.66 Section 640.66 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) BIOLOGICS ADDITIONAL STANDARDS FOR HUMAN BLOOD AND BLOOD PRODUCTS Source Plasma 640.66 Immunization of donors....

  1. 21 CFR 640.31 - Suitability of donors.

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ... 21 Food and Drugs 7 2014-04-01 2014-04-01 false Suitability of donors. 640.31 Section 640.31 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) BIOLOGICS ADDITIONAL STANDARDS FOR HUMAN BLOOD AND BLOOD PRODUCTS Plasma 640.31 Suitability of donors. (a)...

  2. Crowd Around: Expanding Your Donor Pool with Crowdfunding

    ERIC Educational Resources Information Center

    Jarrell, Andrea

    2013-01-01

    At most institutions, annual fund-giving is down. Crowdfunding sites allow people with a great idea or worthy cause to bypass traditional funding methods and take their case directly to web-savvy investors and donors. This article describes how higher education institutions are expanding their donor pool through such crowdfunding sites as USEED,

  3. University of Washington Medical Center Living Donor Program

    E-print Network

    Borenstein, Elhanan

    with important information to help evaluate your potential as living kidney donor also to help and support you of your kidneys? 2. How long have you been contemplating this decision? 3. What do you know at this time about being a living kidney donor? Where did you obtain this information? #12;University of Washington

  4. Fathers Anonymous: Beyond the Best Interests of the Sperm Donor.

    ERIC Educational Resources Information Center

    Annas, George J.

    1981-01-01

    Reviews the rationale for the practice of Artificial Insemination Donor (AID) practices, the manner whereby donors are selected, and how records are kept. The importance of developing standards which serve the best interests of the AID child is stressed. (Author/DB)

  5. 21 CFR 640.63 - Suitability of donor.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ...HUMAN BLOOD AND BLOOD PRODUCTS Source Plasma 640.63 Suitability of donor... The suitability of a donor for Source Plasma shall be determined by a qualified...immunized for the production of high-titer plasma shall be examined by a qualified...

  6. 21 CFR 640.63 - Suitability of donor.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ...HUMAN BLOOD AND BLOOD PRODUCTS Source Plasma 640.63 Suitability of donor... The suitability of a donor for Source Plasma shall be determined by a qualified...immunized for the production of high-titer plasma shall be examined by a qualified...

  7. 21 CFR 640.63 - Suitability of donor.

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ...HUMAN BLOOD AND BLOOD PRODUCTS Source Plasma 640.63 Suitability of donor... The suitability of a donor for Source Plasma shall be determined by a qualified...immunized for the production of high-titer plasma shall be examined by a qualified...

  8. 21 CFR 640.63 - Suitability of donor.

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ...HUMAN BLOOD AND BLOOD PRODUCTS Source Plasma 640.63 Suitability of donor... The suitability of a donor for Source Plasma shall be determined by a qualified...immunized for the production of high-titer plasma shall be examined by a qualified...

  9. 21 CFR 640.63 - Suitability of donor.

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ...HUMAN BLOOD AND BLOOD PRODUCTS Source Plasma 640.63 Suitability of donor... The suitability of a donor for Source Plasma shall be determined by a qualified...immunized for the production of high-titer plasma shall be examined by a qualified...

  10. You...as Blood Donor: Teacher Strategies and Student Worksheets.

    ERIC Educational Resources Information Center

    Degroat, Diane Zagare; And Others

    This curriculum guide for teaching about blood donation was prepared to improve school-community participation in the New York City Blood Donor Program. It contains plans for five lessons on the following topics: (1) the nature of blood; (2) blood and technology--modern-day advances; (3) blood and your personal health; (4) the blood donor as good

  11. Hyperfine Stark effect of shallow donors in silicon

    NASA Astrophysics Data System (ADS)

    Pica, Giuseppe; Wolfowicz, Gary; Urdampilleta, Matias; Thewalt, Mike L. W.; Riemann, Helge; Abrosimov, Nikolai V.; Becker, Peter; Pohl, Hans-Joachim; Morton, John J. L.; Bhatt, R. N.; Lyon, S. A.; Lovett, Brendon W.

    2014-11-01

    We present a complete theoretical treatment of Stark effects in bulk doped silicon, whose predictions are supported by experimental measurements. A multivalley effective mass theory, dealing nonperturbatively with valley-orbit interactions induced by a donor-dependent central cell potential, allows us to obtain a very reliable picture of the donor wave function within a relatively simple framework. Variational optimization of the 1 s donor binding energies calculated with a new trial wave function, in a pseudopotential with two fitting parameters, allows an accurate match of the experimentally determined donor energy levels, while the correct limiting behavior for the electronic density, both close to and far from each impurity nucleus, is captured by fitting the measured contact hyperfine coupling between the donor nuclear and electron spin. We go on to include an external uniform electric field in order to model Stark physics: with no extra ad hoc parameters, variational minimization of the complete donor ground energy allows a quantitative description of the field-induced reduction of electronic density at each impurity nucleus. Detailed comparisons with experimental values for the shifts of the contact hyperfine coupling reveal very close agreement for all the donors measured (P, As, Sb, and Bi). Finally, we estimate field ionization thresholds for the donor ground states, thus setting upper limits to the gate manipulation times for single qubit operations in Kane-like architectures: the Si:Bi system is shown to allow for A gates as fast as ?10 MHz.

  12. Non-local coupling of two donor-bound electrons

    NASA Astrophysics Data System (ADS)

    Verduijn, J.; Agundez, R. R.; Blaauboer, M.; Rogge, S.

    2013-03-01

    We report the results of an experiment investigating coherence and correlation effects in a system of coupled donors. Two donors are strongly coupled to two leads in a parallel configuration within a nano-wire field effect transistor. By applying a magnetic field we observe interference between two donor-induced Kondo channels, which depends on the Aharonov-Bohm phase picked up by electrons traversing the structure. This results in a non-monotonic conductance as a function of magnetic field and clearly demonstrates that donors can be coupled through a many-body state in a coherent manner. We present a model which shows good qualitative agreement with our data. The presented results add to the general understanding of interference effects in a donor-based correlated system which may allow us to create artificial lattices that exhibit exotic many-body excitations.

  13. Using fertile couples as embryo donors: An ethical dilemma

    PubMed Central

    Alizadeh, Leila; Omani Samani, Reza

    2014-01-01

    The use of donated embryos has offered hope for infertile couples who have no other means to have children. In Iran, fertility centers use fertile couples as embryo donors. In this paper, the advantages and disadvantages of this procedure will be discussed. We conclude that embryo-donation should be performed with frozen embryos thus preventing healthy donors from being harmed by fertility drugs. There must be guidelines for choosing the appropriate donor families. In countries where commercial egg donation is acceptable, fertile couples can be procured as embryo donors thus fulfilling the possible shortage of good quality embryos. Using frozen embryos seems to have less ethical, religious and legal problems when compared to the use of fertile embryo donors. PMID:24799876

  14. Adiabatic Charge Control in a Single Donor Atom Transistor

    E-print Network

    Enrico Prati; Matteo Belli; Simone Cocco; Guido Petretto; Marco Fanciulli

    2010-08-10

    We charge an individual donor with electrons stored in a quantum dot in its proximity. A Silicon quantum device containing a single Arsenic donor and an electrostatic quantum dot in parallel is realized in a nanometric field effect transistor. The different coupling capacitances of the donor and the quantum dot with the control and the back gates are exploited to generate a relative rigid shift of their energy spectrum as a function of the back gate voltage, causing the crossing of the energy levels. We observe the sequential tunneling through the $D^{2-}$ and the $D^{3-}$ energy levels of the donor hybridized at the oxide interface at 4.2 K. Their respective states form an honeycomb pattern with the quantum dot states. It is therefore possible to control the exchange coupling of an electron of the quantum dot with the electrons bound to the donor, thus realizing a physical qubit for quantum information processing applications.

  15. The Significance of Evalution of Haematocrit in Plateletpheresis Donors

    PubMed Central

    Jagtap, Pratap Eknath; Nagane, Nitin Sopan; Dhonde, Sushma Prakash; Belwalkar, Gajanan J.; Mane, Vaibhav Pandurang

    2015-01-01

    Background: The collection of platelets by apheresis is considered as a very great progress in transfusion medicine. In present era, many automated cell separation are available each model has tried to improve productivity, quality of plateletpheresis. Further various studies have been done to correlate the quality of platelet concentrates. Also, various biochemical studies have been done on plateletpheresis donors. However, safety issue with regards to post procedure levels of biochemical parameters decreased in donors undergoing plateletpheresis have been only minimally explored. Objectives: Investigating Haematological and Biochemical parameters (Hematocrit value and Serum Calcium levels) pre and post in plateletpheresis donors. Materials and Methods: Sixty two healthy first time voluntary plateletpheresis donors at Apheresis unit in blood bank Bharati Vidyapeeth Deemed University Medical College & Hospital, Sangli, Maharashtra, India. Hematocrit value of plateletpheresis donors were analysed and based on mean value 43.2% considering this as standard in the present study. We categorized plateletpheresis donors in two groups (A) these having value less than 43.2% (n = 36) and Group (B) having haematocrit more than 43.3% (n = 26). Volume of ACD required for donors from both group were noted. Result: We observed mean of ACD infused in group A plateletpheresis donors was 347.7 ml 35.75 SD while group 態 donors required mean volume ACD to be infused was 379.6 ml 46.24 S.D. was statistically significant (p <0.005). Conclusion: Plateletpheresis induces marked metabolic effects, with sustained changes in serum calcium and haematocrit after ACD infusion, the results show, before procedure (Plateletpheresis) one must consider the haematocrit value along with serum calcium levels in Plateletpheresis donor to avoid severe symptoms of hypocalcaemia. PMID:26023547

  16. Pushing the frontiers of living donor right hepatectomy

    PubMed Central

    Kim, Seong Hoon; Lee, Seung Duk; Kim, Young Kyu; Park, Sang-Jae

    2014-01-01

    Living donor right hepatectomy (LDRH) is currently the most common donor surgery in adult-to-adult living donor liver transplantation although the morbidity and mortality reported in living donors still contradicts the Hippocratic tenet of 揹o no harm. Achieving low complication rates in LDRH remains a matter of major concern. Living donor surgery is performed worldwide as an established solution to the donor shortage. The aim of this study was to assess the current status of LDRH and comment on the future of the procedure; assessment was made from the standpoint of optimizing the donor selection criteria and reducing morbidity based on both the authors 8-year institutional experience and a literature review. New possibilities have been explored regarding selection criteria. The safety of living donors with unfavorable conditions, such as low remnant liver volume, fatty change, or old age, should also be considered. Abdominal incisions have become shorter, even without laparoscopic assistance; upper midline laparotomy is the primary incision used in more than 400 consecutive LDRHs in the authors institution. Various surgical techniques based on preoperative imaging technology of vascular and biliary anomalies have decreased the anatomical barriers in LDRH. Operative time has been reduced, with low blood loss. Laparoscopic or robotic LDRH has been tried in only a few selected donors. The LDRH-specific, long-term outcomes remain to be addressed. The follow-up duration of these studies should be long enough to address possible late complications. Donor safety, which is the highest priority, is ensured by three factors: preoperative selection, intraoperative surgical technique, and postoperative management. These three focus areas should be continuously refined, with the ultimate goal of zero morbidity. PMID:25561777

  17. Update on donor assessment, resuscitation, and acceptance criteria, including novel techniques--non-heart-beating donor lung retrieval and ex vivo donor lung perfusion.

    PubMed

    Yeung, Jonathan C; Cypel, Marcelo; Waddell, Thomas K; van Raemdonck, Dirk; Keshavjee, Shaf

    2009-05-01

    The shortage of adequate organ donors remains a great challenge in clinical lung transplantation. With increasing experience in the medical management and surgical technique of lung transplantation, gradual expansion of the criteria for lung donor selection has occurred with beneficial effects on the donor pool. Interest in donation after cardiac death also is increasing as the gap increases between donors and the needs of listed patients. Successful use of these new sources of lungs depends on the accurate assessment and prediction of transplanted lung function. Promising techniques for lung assessment and diagnostics include investigating key genes associated with graft failure or good graft performance using molecular approaches, and ex vivo evaluation. Further studies are needed to answer remaining questions about the best technique and solution to reperfuse human lungs for several hours without edema formation. As the predictive ability to discern good from injured donor lungs improves, strategies to repair donor lungs become increasingly important. Prolonged normothermic EVLP seems to be a platform on which many reparative strategies can be realized. With these new methods for assessing and resuscitating lungs accurately, it is hoped that inroads will be made toward providing every listed patient a chance for successful lung transplantation. PMID:19662970

  18. Highly sensitive immunoassay of protein molecules based on single nanoparticle fluorescence detection in a nanowell

    NASA Astrophysics Data System (ADS)

    Han, Jin-Hee; Kim, Hee-Joo; Lakshmana, Sudheendra; Gee, Shirley J.; Hammock, Bruce D.; Kennedy, Ian M.

    2011-03-01

    A nanoarray based-single molecule detection system was developed for detecting proteins with extremely high sensitivity. The nanoarray was able to effectively trap nanoparticles conjugated with biological sample into nanowells by integrating with an electrophoretic particle entrapment system (EPES). The nanoarray/EPES is superior to other biosensor using immunoassays in terms of saving the amounts of biological solution and enhancing kinetics of antibody binding due to reduced steric hindrance from the neighboring biological molecules. The nanoarray patterned onto a layer of PMMA and LOL on conductive and transparent indium tin oxide (ITO)-glass slide by using e-beam lithography. The suspension of 500 nm-fluorescent (green emission)-carboxylated polystyrene (PS) particles coated with protein-A followed by BDE 47 polyclonal antibody was added to the chip that was connected to the positive voltage. The droplet was covered by another ITO-coated-glass slide and connected to a ground terminal. After trapping the particles into the nanowells, the solution of different concentrations of anti-rabbit- IgG labeled with Alexa 532 was added for an immunoassay. A single molecule detection system could quantify the anti-rabbit IgG down to atto-mole level by counting photons emitted from the fluorescent dye bound to a single nanoparticle in a nanowell.

  19. Ultrasensitive on-chip immunoassays with a nanoparticle-assembled photonic crystal.

    PubMed

    Han, Jin-Hee; Sudheendra, L; Kim, Hee-Joo; Gee, Shirley J; Hammock, Bruce D; Kennedy, Ian M

    2012-10-23

    Electrophoretic particle entrapment system (EPES) is employed to generate 2D array of nanoparticles coated with biological molecules (i.e., antibodies). Phase matching of the excitation and the emission in the 2D arrays with particles produces a highly enhanced fluorescence signal that was shown to improve the limit of detection in immunoassays. The phase matching is achieved when the particle are in the sub-100 nm range. A comparison between different size particles shows that the sensitivity of an immunoassay is extended to a range that is difficult to achieve with standard technology (e.g., enzyme-linked immunosorbent assay-ELISA). The effectiveness of this novel configuration of particle-in-a-well was demonstrated with an assay for human epidermal growth factor receptor 2 (HER2; breast cancer biomarker), with a detection limit as low as 10 attomolar (aM) in less than 10 ?L of serum-based sample. The limit of detection of HER2 indicated far superior assay performance compared to the corresponding standard 96-well plate-based ELISA. The particle-based photonic platform reduces the reagent volume and the time for performing an assay in comparison to competing methods. The simplicity of operation and the level of sensitivity demonstrated here can be used for rapid and early stage detection of biomarkers. PMID:22957818

  20. Development of sensitive direct chemiluminescent enzyme immunoassay for the determination of dihydroartemisinin in plasma.

    PubMed

    Zehnacker, Laura; Nevers, Marie-Claire; Sinou, V閞onique; Parzy, Dominique; Cr閙inon, Christophe; Parzy, Daniel; Azoulay, St閜hane

    2015-10-01

    Despite significant progress in prevention and therapy, malaria is still one of the world's leading major diseases due to its high morbidity and mortality. Recommended treatments by the World Health Organization include the use of artemisinin and artemisinin derivative-based combination therapies. To allow efficient patient monitoring during antimalarial therapy without the use of expensive apparatus, we developed a sensitive direct chemiluminescent enzyme immunoassay for the determination of dihydroartemisinin in biological fluids. To produce specific antibodies against dihydroartemisinin (DHA), a synthetic DHA derivative was coupled to bovine serum albumin as the immunogen. In parallel, a new, rapid, and efficient procedure to covalently link glycoprotein to all amine-containing molecules has been established and the enzyme tracer was prepared by chemically coupling the DHA derivative in combination with SBP rather than the more commonly used HRP. It allowed us to develop, after optimization of the luminescent reagent, a sensitive and stable luminescent EIA, with a LLOQ of 90 pg mL(-1). This assay compares favorably with the most efficient HPLC methods previously reported with a LLOQ close to 1 ng mL(-1) and shows good precision and efficiency since recovery from human plasma spiked with DHA ranged between 91 and 103%, with coefficients of variation of <13%. To date, no immunoassay for DHA has been applied to plasma analysis and this EIA should be very useful in all clinical laboratories for rapid and cost-effective analysis. PMID:26280205

  1. Establishment of immunoassay for detecting HPV16 E6 and E7 RNA

    PubMed Central

    Ding, Sen; Qian, Steven Y.; Zhang, Yang; Wu, Wenlei; Lu, Gensheng; Lu, Yan; Feng, Xiujing; Li, Li; Shen, Pingping

    2015-01-01

    Cervical carcinoma is the most prevalent malignancy second only to breast cancer among women worldwide. Since more than 99% of cervical cancers are caused by human papilloma virus (HPV), measurement of HPV (HPV test) was commonly used in screening risk and/or early stage of cervical cancer as well as assessing the efficacies of the treatments that can decrease the incidence of cervical cancer. Many approaches that diagnose HPV infections have been developed, while most of them have distinct shortcomings. We here established a novel immunoassay method in which the pairs of unlabeled DNA probes firstly bind to HPV16 E6 and E7 RNAs to form the DNA-RNA hybrids, and the hybrids will subsequently be identified by S9.6 antibody. The sensitivity of this highly specific method can reach ~0.923?pg/mL and ~0.424?pg/mL of in vitro transcribed HPV16 E6 and E7 RNA, respectively, and reverse transcription and polymerase chain reaction (PCR) amplification were no longer needed. Thus, our immunoassay approaches can precisely reflect the actually viral load that is related to the course of HPV infection. In addition, it has also fast and low cost characteristic feature. PMID:26333509

  2. Magnetic Bead-Based Colorimetric Immunoassay for Aflatoxin B1 Using Gold Nanoparticles

    PubMed Central

    Wang, Xu; Niessner, Reinhard; Knopp, Dietmar

    2014-01-01

    A competitive colorimetric immunoassay for the detection of aflatoxin B1 (AFB) has been established using biofunctionalized magnetic beads (MBs) and gold nanoparticles (GNPs). Aflatoxin B1-bovine serum albumin conjugates (AFB-BSA) modified MBs were employed as capture probe, which could specifically bind with GNP-labeled anti-AFB antibodies through immunoreaction, while such specific binding was competitively inhibited by the addition of AFB. After magnetic separation, the supernatant solution containing unbound GNPs was directly tested by UV-Vis spectroscopy. The absorption intensity was directly proportional to the AFB concentration. The influence of GNP size, incubation time and pH was investigated in detail. After optimization, the developed method could detect AFB in a linear range from 20 to 800 ng/L, with the limit of detection at 12 ng/L. The recoveries for spiked maize samples ranged from 92.8% to 122.0%. The proposed immunoassay provides a promising approach for simple, rapid, specific and cost-effective detection of toxins in the field of food safety. PMID:25405511

  3. Detection of inflammatory cytokines using a fiber optic microsphere immunoassay array

    NASA Astrophysics Data System (ADS)

    Blicharz, Timothy M.; Walt, David R.

    2006-10-01

    A multiplexed fiber optic microsphere-based immunoassay array capable of simultaneously measuring five inflammatory cytokines has been developed. Five groups of amine-functionalized 3.1 micron microspheres were internally encoded with five distinct concentrations of a europium dye and converted to cytokine probes by covalently coupling monoclonal capture antibodies specific for human VEGF, IFN-gamma, RANTES, IP-10, and Eotaxin-3 to the microspheres via glutaraldehyde chemistry. The microspheres were pooled and loaded into a 1 mm diameter fiber optic bundle containing ~50,000 individual etched microwells, producing the multiplexed cytokine immunoassay array. Multiple arrays can be created from a single microsphere pool for high throughput sample analysis. Sandwich fluoroimmunoassays were performed by incubating the probe array in a sample, followed by incubation in a mixture of biotin-labeled detection antibodies that are complementary to the five cytokines. Finally, universal detection of each protein was performed using a fluorescence imaging system after briefly immersing the array in a solution of fluorophore-labeled streptavidin. The multiplexed cytokine array has been shown to respond selectively to VEGF, IFNgamma, RANTES, IP-10, and Eotaxin-3, permitting multiplexed quantitative analysis. Ultimately, the multiplexed cytokine array will be utilized to evaluate the potential of using saliva as a noninvasive diagnostic fluid for pulmonary inflammatory diseases such as asthma.

  4. Detection of one milliattomole of ferritin by novel and ultrasensitive enzyme immunoassay.

    PubMed

    Hashida, S; Ishikawa, E

    1990-12-01

    A novel and ultrasensitive enzyme immunoassay (immune complex transfer two-site enzyme immunoassay) for ferritin is described. Ferritin was reacted simultaneously with affinity-purified dinitrophenyl biotinyl anti-ferritin IgG and affinity-purified anti-ferritin Fab'-beta-D-galactosidase conjugate. The complex formed of the three components was trapped onto affinity-purified (anti-dinitrophenyl group) IgG-coated polystyrene balls. After eliminating excess conjugate by washing, the complex was eluted from the polystyrene balls with an excess of epsilon N-dinitrophenyl-L-lysine and transferred to streptavidin-coated polystyrene balls. The beta-D-galactosidase activity bound to streptavidin-coated polystyrene balls was assayed by fluorometry. Nonspecifically bound beta-D-galactosidase activity was remarkably lowered but there was much less decrease in specifically bound beta-D-galactosidase activity. As a result, the detection limit of ferritin was lowered to 1 milliattomole (1 x 10(-21) mol, 600 molecules as calculated from Avogadro's number). This technique will be useful for measuring, for example, antigens in single cells. PMID:2128491

  5. Protein immunoassay methods for detection of biotech crops: applications, limitations, and practical considerations.

    PubMed

    Stave, James W

    2002-01-01

    Immunoassay methods are available for detection and quantitation of proteins expressed by most biotechnology-derived crops in commercial production. The 2 most common test formats are enzyme-linked immunosorbent assay (ELISA) and immunochromatographic (lateral flow) strip tests. Two ELISA methods, one for Roundup Ready soybeans and one for MON810 CrylAb corn, were the subject of large international collaborative studies and were demonstrated to quantitatively determine the concentrations of biotech crops in samples of ground grain. Quantitative ELISA methods are also useful for analysis of processed fractions of agricultural commodities such as soybean toasted meal or corn flour. Both strip tests and ELISAs for biotech crops are currently being used on a large scale in the United States to manage the sale and distribution of grain. In these applications, tests are used to determine if the concentration of biotech grain is above or below specified threshold limits. Using existing U.S. Department of Agriculture sampling techniques, the reliability of the threshold determination is expressed in terms of statistical confidence rather than analytical precision. Combining the use of protein immunoassays with Identity Preservation systems provides an effective means of characterizing the raw and processed agricultural inputs to the food production system in a way that allows food producers to comply with labeling laws. PMID:12083275

  6. Photoelectrochemical detection of enzymatically generated CdS nanoparticles: Application to development of immunoassay.

    PubMed

    Barroso, Javier; Saa, Laura; Grinyte, Ruta; Pavlov, Valeri

    2016-03-15

    We report an innovative photoelectrochemical process (PEC) based on graphite electrode modified with electroactive polyvinylpyridine bearing osmium complex (Os-PVP). The system relies on the in situ enzymatic generation of CdS quantum dots (QDs). Alkaline phosphatase (ALP) catalyzes the hydrolisis of sodium thiophosphate (TP) to hydrogen sulfide (H2S) which in the presence Cd(2+) ions yields CdS semiconductor nanoparticles (SNPs). Irradiation of SNPs with the standard laboratory UV-illuminator (wavelength of 365nm) results in photooxidation of 1-thioglycerol (TG) mediated by Os-PVP complex on the surface of graphite electrode at applied potential of 0.31V vs. Ag/AgCl. A novel immunoassay based on specific enzyme linked immunosorbent assay (ELISA) combined with the PEC methodology was developed. Having selected the affinity interaction between bovine serum albumine (BSA) with anti-BSA antibody (AB) as a model system, we built the PEC immunoassay for AB. The new assay displays a linear range up to 20ngmL(-1) and a detection limit (DL) of 2ngmL(-1) (S/N=3) which is lower 5 times that of the traditional chromogenic ELISA test employing p-nitro-phenyl phosphate (pNPP). PMID:26432195

  7. Multiplexed In-cell Immunoassay for Same-sample Protein Expression Profiling

    PubMed Central

    Shang, Jing; Zrazhevskiy, Pavel; Postupna, Nadia; Keene, C. Dirk; Montine, Thomas J.; Gao, Xiaohu

    2015-01-01

    In-cell immunoassays have become a valuable tool for protein expression analysis complementary to established assay formats. However, comprehensive molecular characterization of individual specimens has proven challenging and impractical due to, in part, a singleplex nature of reporter enzymes and technical complexity of alternative assay formats. Herein, we describe a simple and robust methodology for multiplexed protein expression profiling on the same intact specimen, employing a well-characterized enzyme alkaline phosphatase for accurate quantification of all targets of interest, while overcoming fundamental limitations of enzyme-based techniques by implementing the DNA-programmed release mechanism for segregation of sub-sets of target-bound reporters. In essence, this methodology converts same-sample multi-target labeling into a set of isolated singleplex measurements performed in a parallel self-consistent fashion. For a proof-of-principle, multiplexed detection of three model proteins was demonstrated on cultured HeLa cells, and two clinically-relevant markers of dementia, ?-amyloid and PHF-tau, were profiled in formalin-fixed paraffin embedded brain tissue sections, uncovering correlated increase in abundance of both markers in the 揂lzheimer抯 disease cohort. Featuring an analytically powerful yet technically simple and robust methodology, multiplexed in-cell immunoassay is expected to enable insightful same-sample protein profiling studies and become broadly adopted in biomedical research and clinical diagnostics. PMID:26328896

  8. Multiple immunoassay systems are negatively interfered by circulating cardiac troponin I autoantibodies.

    PubMed

    Tang, Gusheng; Wu, Yu; Zhao, Weiguo; Shen, Qian

    2012-03-01

    Circulating cardiac troponin I (cTnI) autoantibodies have recently been detected in more and more patients with myocardial injury. In the present study, a total of 121 patients with acute myocardial infarction (AMI) were screened for cTnI autoantibodies using an indirect ELISA. Positive results were further confirmed by Western blot analysis. As a result, 13 autoantibody-positive sera were identified, in which cTnI values detected by different immunoassay systems are very different. Further evaluation revealed low recovery in one of the 13 samples with the Access 2 system (Beckman Coulter, 2^ generation), one low and one moderate recovery sample with Architect i2000 (Abbott), one low and two moderate with AxSYM (Abbott), two low and three moderate with Dimension Xpand (Dade Behring, 2^ generation), and four low and one moderate with Vidas (bioM閞ieux). Our work demonstrates that circulating cTnI autoantibodies occur in part of patients with AMI and, for the first time to our knowledge, shows that these autoantibodies can result in considerable negative interference in all the five commonly used cTnI immunoassay systems, which may lead to incorrect diagnoses and following treatments. The indirect ELISA established in our laboratory is suitable for a rapid preliminary screening for cTnI autoantibody in clinical work. PMID:21655961

  9. Clinical performance evaluation of four automated chemiluminescence immunoassays for hepatitis C virus antibody detection.

    PubMed

    Kim, Sinyoung; Kim, Jeong-Ho; Yoon, Seoyoung; Park, Youn-Hee; Kim, Hyon-Suk

    2008-12-01

    Various automated chemiluminescence immunoassay (CLIA) analyzers for the detection of antibodies to hepatitis C virus (HCV) are now commercially available in clinical laboratories and are replacing conventional enzyme immunoassays. We investigated the performance of four anti-HCV CLIAs (the Architect Anti-HCV assay on the Architect i2000 system, the Vitros Anti-HCV assay on the Vitros ECiQ Immunodiagnostic System, the Access HCV Ab PLUS assay on the UniCel DxI 800 analyzer, and the newly developed Elecsys Anti-HCV assay on the Cobas e 411 analyzer). The total percent coefficient of variation values of imprecision were 3.5 to 5.7% with positive control materials and 7.2 to 10.2% with negative control materials. The agreement between the results of the Elecsys, Architect, Vitros, and Access CLIAs ranged from 94.5 to 98.1%. The clinical sensitivity of all CLIAs was 100%. Each CLIA showed excellent reproducibility and clinical sensitivity. The Elecsys, Architect, Vitros, and Access CLIAs showed clinical specificities of 98.2, 98.8, 96.5, and 98.2%. PMID:18945839

  10. Plasma exchange to remove HIT antibodies: dissociation between enzyme-immunoassay and platelet activation test reactivities.

    PubMed

    Warkentin, Theodore E; Sheppard, Jo-Ann I; Chu, F Victor; Kapoor, Anil; Crowther, Mark A; Gangji, Azim

    2015-01-01

    Repeated therapeutic plasma exchange (TPE) has been advocated to remove heparin-induced thrombocytopenia (HIT) IgG antibodies before cardiac/vascular surgery in patients who have serologically-confirmed acute or subacute HIT; for this situation, a negative platelet activation assay (eg, platelet serotonin-release assay [SRA]) has been recommended as the target serological end point to permit safe surgery. We compared reactivities in the SRA and an anti-PF4/heparin IgG-specific enzyme immunoassay (EIA), testing serial serum samples in a patient with recent (subacute) HIT who underwent serial TPE precardiac surgery, as well as for 15 other serially-diluted HIT sera. We observed that post-TPE/diluted HIT sera-when first testing SRA-negative-continue to test strongly positive by EIA-IgG. This dissociation between the platelet activation assay and a PF4-dependent immunoassay for HIT antibodies indicates that patients with subacute HIT undergoing repeated TPE before heparin reexposure should be tested by serial platelet activation assays even when their EIAs remain strongly positive. PMID:25406354

  11. Immunoassay for detection of antibodies to simian immunodeficiency virus and human immunodeficiency virus in serum.

    PubMed

    Otsyula, M G; Yee, J A; Suleman, M A; Marx, P A; Jennings, M B

    1996-04-01

    We developed a simple, inexpensive, rapid assay for the detection of antibodies to simian immunodeficiency virus (SIV) and human immunodeficiency virus type 1 (HIV-1) in serum. The immunoassay uses inactivated SIV and HIV-1 gp41 transmembrane recombinant protein as antigenic adsorbents on a nitrocellulose filter membrane. Diluted serum, with the addition of Protein-A-Gold, is gravity-filtered through the filter membrane, blocked, and buffer-washed. Antibodies to HIV or SIV or both in serum bind to the appropriate antigen, and the resulting antigen-antibody complex reacts with Protein-A-Gold to produce a readable pink color. Field evaluation of the test on 30 human and 70 nonhuman primate sera in Kenya and Zaire indicated that the test had at least 93 and 90% correlation with Western blot sensitivity and specificity respectively. Prior refrigeration of the test kit and incubation of sera during testing were not required. This result indicates that the test may be a rapid, economical, and simple test for detecting HIV, SIV, or both in serum. This immunoassay can be useful for carrying out HIV and SIV serosurveys in countries with limited or no laboratory facilities. PMID:8723237

  12. REVIEW ARTICLE: Trends in interfacial design for surface plasmon resonance based immunoassays

    NASA Astrophysics Data System (ADS)

    Shankaran, Dhesingh Ravi; Miura, Norio

    2007-12-01

    Immunosensors based on surface plasmon resonance (SPR) have become a promising tool in sensor technology for biomedical, food, environmental, industrial and homeland security applications. SPR is a surface sensitive optical technique, suitable for real-time and label-free analysis of biorecognition events at functional transducer surfaces. Fabrication of highly active and robust sensing surfaces is an important part in immunoassays because the quality, quantity, chemistry and topography of the interfacial biomembranes play a major role in immunosensor performance. Eventually, a variety of immobilization methods such as physical adsorption, covalent coupling, Langmuir Blodgett film, polymer thin film, self-assembly, sol gel, etc, have been introduced over the years for the immobilization of biomolecules (antibody or antigen) on the transducer surfaces. The selection of an immobilization method for an immunoassay is governed by several factors such as nature and stability of the biomolecules, target analyte, application, detection principle, mode of signal transduction, matrix complexity, etc. This paper provides an overview of the various surface modification methods for SPR based immunosensor fabrication. The preparation, structure and application of different functional interfacial surfaces have been discussed along with a brief introduction to the SPR technology, biomolecules and detection principles.

  13. Ultrasensitive On-Chip Immunoassays with a Nanoparticle-Assembled Photonic Crystal

    PubMed Central

    Han, Jin-Hee; Sudheendra, L.; Kim, Hee-Joo; Gee, Shirley J.; Hammock, Bruce D.; Kennedy, Ian M.

    2012-01-01

    Electrophoretic particle entrapment system (EPES) is employed to generate 2D array of nanoparticles coated with biological molecules (i.e. antibodies). Phase matching of the excitation and the emission in the 2D arrays with particles produces a highly enhanced fluorescence signal that was shown to improve the limit of detection in immunoassays. The phase matching is achieved when the particle are in the sub-100 nm range. A comparison between different size particles shows that the sensitivity of an immunoassay is extended to a range that is difficult to achieve with standard technology (e.g. Enzyme-linked immunosorbent assay-ELISA). The effectiveness of this novel configuration of particle-in-a well was demonstrated with an assay for human epidermal growth factor receptor 2 (HER2; breast cancer biomarker), with a detection limit as low as 10 aM in less than 10 ?l of serum-based sample. The limit of detection of HER2 indicated far superior assay performance compared to the corresponding standard 96-well plate-based ELISA. The particle-based photonic platform reduces the reagent volume and the time for performing an assay in comparison to competing methods. The simplicity of operation and the level of sensitivity demonstrated here can be used for rapid and early-stage detection of biomarkers. PMID:22957818

  14. Validation of multiplex microbead immunoassay for simultaneous serodetection of multiple infectious agents in laboratory mouse.

    PubMed

    Ravindran, Resmi; Khan, Imran H; Krishnan, Viswanathan V; Ziman, Melanie; Kendall, Lon V; Frasier, Janelle M; Bates, Rachel; Griffey, Steve M; Fahey, James R; Luciw, Paul A

    2010-12-15

    Multiplex methodologies enable simultaneous detection of antibodies against several infectious agents allowing sample conservation, cost effectiveness, and amenability to high-throughput/automation. We have previously described a multiplex microbead immunoassay for serodetection of ten, high-priority mouse infectious pathogens. Here, we present a validation of this multiplex diagnostic system using approximately four hundred serum samples from different groups of mice. Computer assisted multivariate analysis of the resulting high volume data (8000 data points) was performed. This computational approach enabled presentation of data in a variety of easily interpretable formats (e.g., correlation tables and heat maps). Importantly, this computer aided approach was instrumental for the evaluation of assay accuracy, sensitivity, specificity, and robustness during the study. Crucial pieces of information were obtained to make timely adjustments for assay refinement. This progressive approach to developing an implementation-ready clinical assay, facilitated by computational analysis, produced a highly efficient, accurate and dependable serodiagnostics system. This system has effectively replaced the current state-of-the-art methodology (ELISA) used in mouse colony health management at the University of California and the Jackson Laboratory. A pathway to develop multiplex serology tests for infectious disease diagnosis described here serves as a model for multiplex immunoassay design, clinical validation, refinement and implementation. PMID:20965193

  15. Accelerated immunoassays based on magnetic particle dynamics in a rotating capillary tube with stationary magnetic field

    PubMed Central

    Lee, Jun-Tae; Sudheendra, L.; Kennedy, Ian M.

    2012-01-01

    A rapid and simple magnetic particle-based immunoassay has been demonstrated in a capillary mixing system. Antibody-coated micrometer size super-paramagnetic polystyrene (SPP) particles were used in an assay for rabbit IgG in a sandwich (non-competitive) format. The kinetics of the assay was compared between a plate based system and a single capillary tube. The interaction between the antigen (R-IgG) and the antibody (anti-R-IgG) that was carried by the SPP particles in a rotating capillary was tested under a stationary magnetic field. Competing magnetic and viscous drag forces helped to enhance the interaction between the analyte and the capture antibodies on the particles. The dimensionless Mason number (Mn) was employed to characterize the magnetic particle dynamics a previously determined critical Mason number (Mnc) was employed as a guide to the appropriate experimental conditions of magnetic field strength and rotational speed of the capillary. The advantage of the rotating capillary system included a short assay time and a reduced reactive volume (20?l). The results show that the immunoassay kinetics were improved by the formation of chains of the SPP particles for the conditions that corresponded to the critical Mason number. PMID:22931580

  16. A reusable biosensor chip for SERS-fluorescence dual mode immunoassay

    NASA Astrophysics Data System (ADS)

    Wu, Lei; Wang, Zhuyuan; Fan, Kequan; Zong, Shenfei; Cui, Yiping

    2015-05-01

    Research continues in an effort to develop a versatile platform for clinical diagnosis with easy operation and low cost. In the present study, a biosensor chip has been designed and fabricated for surface enhanced Raman scattering (SERS)- fluorescence dual mode immunoassay. Here, a dual channel microfluidic chip was employed for simultaneous SERS and fluorescence detection. Unlike previously reported microfluidic immunoassays using fluorescence or SERS method independently, the proposed dual mode biosensor combined the advantages of these two optical detection techniques. The fluorescence mode can be used for fast screening of biomolecules while the SERS mode can be employed for accurate and sensitive quantitative analysis. In addition, the chip-based microfluidic platform greatly reduced the reagents cost and complicated operation. The whole detection process from sample preparation to optical detection can be finished in 90 min. Moreover, the reversibly bonded biosensor chip could be reused after cleaning, which further reduced the cost for each assay. All these merits make it a potential powerful tool for practical clinical diagnosis.

  17. Highly sensitive detection of clenbuterol using competitive surface-enhanced Raman scattering immunoassay.

    PubMed

    Zhu, Guichi; Hu, Yongjun; Gao, Jiao; Zhong, Liang

    2011-07-01

    In this report, we present a novel approach to detect clenbuterol based on competitive surface-enhanced Raman scattering (SERS) immunoassay. Herein, a SERS nanoprobe that relies on gold nanoparticle (GNP) is labeled by 4,4'-dipyridyl (DP) and clenbuterol antibody, respectively. The detection of clenbuterol is carried out by competitive binding between free clenbuterol and clenbuterol-BSA fastened on the substrate with their antibody labeled on SERS nanoprobes. The present method allows us to detect clenbuterol over a much wider concentration range (0.1-100 pg mL(-1)) with a lower limit of detection (ca. 0.1 pg mL(-1)) than the conventional methods. Furthermore, by the use of this new competitive SERS immunoassay, the clenbuterol-BSA (antigen) is chosen to fasten on the substrate instead of the clenbuterol antibody, which could reduce the cost of the assay. Results demonstrate that the proposed method has the wide potential applications in food safety and agonist control. PMID:21641419

  18. 42 CFR 482.102 - Condition of participation: Patient and living donor rights.

    Code of Federal Regulations, 2011 CFR

    2011-10-01

    ...Patient and living donor rights...CONTINUED) STANDARDS AND CERTIFICATION...patient's and living donor's rights. (a) Standard: Informed consent...Part B. (b) Standard: Informed consent for living donors....

  19. 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

    ...Hemoglobin Standards and Maintaining Adequate Iron Stores in Blood Donors; Public Workshop...Hemoglobin Standards and Maintaining Adequate Iron Stores in Blood Donors.'' The purpose...potential measures to maintain adequate iron stores in blood donors. The public...

  20. Will removal of anonymity influence the recruitment of egg donors? A survey of past donors and recipients.

    PubMed

    Craft, Ian; Flyckt, Sofia; Heeley, Gosia; Layland, Sarah; Thornhill, Alan; Kelada, Ehab

    2005-03-01

    The UK Government has determined that children born from egg donation have the same right to know their biological parent as adopted children on reaching the age of 18. There is concern as to the effect of loss of anonymity on egg donor recruitment, since a wait of 1 or 2 years is not unusual for couples awaiting treatment. Some fertility programmes have introduced egg-sharing to circumvent these delays. However, egg-sharing may involve a sub-fertile donor and thus is likely to be less successful overall than egg donation. Recently, the Human Fertilisation and Embryology Authority announced a consultation exercise to consider whether egg donors should receive increased payment or benefits in kind for their donation. To investigate whether or not removal of anonymity would have influenced or would influence their future participation, past egg donors and recipients have been surveyed. Questionnaires were sent to 867 former egg donors (n=504) and recipients (n=363), who were anonymous to each other. Response rates were similar between donors (32.7%) and recipients (39.1%). Of donors, 36.4% would not have participated had donor anonymity been waived, but 69.1% would donate anonymously again. Of recipients, 53.5% would not have proceeded had donor anonymity been waived, whereas 96.5% would receive anonymously donated eggs again. The results indicate that removal of anonymity for egg donors is likely to lead to a further restriction of an already unsatisfactory service to patients requiring donated eggs unless fundamental new initiatives are implemented. PMID:15820037

  1. 76 FR 44013 - Draft Guidance for Industry: Implementation of Acceptable Full-Length and Abbreviated Donor...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2011-07-22

    ...consistent with FDA's requirements and recommendations for collecting Source Plasma donor history information. The Plasma Protein Therapeutics Association (PPTA) Source Plasma donor history questionnaires and accompanying materials (SPDHQ...

  2. Improvements in kidney transplantation from donors after cardiac death.

    PubMed

    Hoogland, E R Pieter; Snoeijs, Maarten G J; Habets, Margot A W; Brandsma, D Steven; Peutz-Kootstra, Carine J; Christiaans, Maarten H L; van Heurn, L W Ernest

    2013-01-01

    To reduce the growing waiting list for kidney transplantation, we explored the limits of kidney transplantation from donors after cardiac death by liberally accepting marginal donor kidneys for transplantation. As the percentage of primary non-function (PNF) increased, we evaluated our transplantation program and implemented changes to reduce the high percentage of PNF in 2005, followed by a second evaluation over the period 2006-2009. Recipients of a kidney from a donor after cardiac death between 1998 and 2005 were analyzed, with PNF as outcome measure. During the period 2002-2005, the percentage of PNF increased and crossed the upper control limits of 12% which was considered as unacceptably high. After implementation of changes, this percentage was reduced to 5%, without changing the number of kidney transplantations from donors after cardiac death. Continuous monitoring of the quality of care is essential as the boundaries of organ donation and transplantation are sought. Meticulous donor, preservation, and recipient management make extension of the donor potential possible, with good results for the individual recipient. Liberal use of kidneys from donors after cardiac death may contribute to a reduction in the waiting list for kidney transplantation and dialysis associated mortality. PMID:23464536

  3. Progress in donor assisted coal liquefaction: Hydroaromatic compound formation

    SciTech Connect

    Kottenstette, R.J.; Stephens, H.P.

    1993-12-31

    The role of hydrogen donor compounds in coal liquefaction has been extensively investigated since the mid 1960`s using model compounds and process derived hydrogen donor solvents. Our recent research and that of other investigators have shown that two model compounds in particular have great efficacy in solvating low rank coals. 1,2,3,10b tetrahydrofluoranthene (H{sub 4}Fl) and 1,2,3,6,7,8 hexahydropyrene (H{sub 6}Py) have been used to dissolve Wyodak coal to > 95% soluble material as measured by tetrahydrofuran (THF). Although these hydrogen donors are very effective, they may not be found in any significant concentrations in actual liquefaction process recycle solvents. Therefore, studies with process derived recycle materials are necessary to understand donor solvent chemistry. The objective of this paper is to present results of solvent hydrogenation experiments using heavy distillate solvents produced during testing at the Wilsonville Advanced Coal Liquefaction Test Facility. We evaluated the impact of hydrogenation conditions upon hydrogen donor formation in process derived distillates and compared these process derived solvents with the highly effective H{sub 4}Fl and H{sub 6}Py donors in coal liquefaction tests. This paper presents data on reaction conditions used for distillate hydrotreating and subsequent coal liquefaction, with an aim toward understanding the relationship between reaction conditions and donor solvent quality in recycle distillates.

  4. Identifying potential kidney donors using social networking web sites.

    PubMed

    Chang, Alexander; Anderson, Emily E; Turner, Hang T; Shoham, David; Hou, Susan H; Grams, Morgan

    2013-01-01

    Social networking sites like Facebook may be a powerful tool for increasing rates of live kidney donation. They allow for wide dissemination of information and discussion and could lessen anxiety associated with a face-to-face request for donation. However, sparse data exist on the use of social media for this purpose. We searched Facebook, the most popular social networking site, for publicly available English-language pages seeking kidney donors for a specific individual, abstracting information on the potential recipient, characteristics of the page itself, and whether potential donors were tested. In the 91 pages meeting inclusion criteria, the mean age of potential recipients was 37 (range: 2-69); 88% were US residents. Other posted information included the individual's photograph (76%), blood type (64%), cause of kidney disease (43%), and location (71%). Thirty-two percent of pages reported having potential donors tested, and 10% reported receiving a live-donor kidney transplant. Those reporting donor testing shared more potential recipient characteristics, provided more information about transplantation, and had higher page traffic. Facebook is already being used to identify potential kidney donors. Future studies should focus on how to safely, ethically, and effectively use social networking sites to inform potential donors and potentially expand live kidney donation. PMID:23600791

  5. Efficacy and Side Effects of Granulocyte Collection in Healthy Donors

    PubMed Central

    Brockmann, Franziska; Kramer, Michael; Bornh鋟ser, Martin; Ehninger, Gerhard; H鰈ig, Kristina

    2013-01-01

    Summary Background We report on the efficacy and side effects of granulocyte collection, which is comparatively infrequently performed in Germany. Methods Data from 378 healthy donors who underwent 914 granulocyte collections between 1999 and 2007 were retrospectively analyzed. Donors received G-CSF (lenograstim) at a median dose of 5.58 (3.257.36) ?g/kg body weight with (n = 243) or without (n = 57) 4 mg dexamethasone. Side effects were recorded by donor monitoring and interview (questionnaire). Results The median granulocyte yield in apheresis products was 8.47 1010 (3.0714.92 1010). Granulocyte yields correlated significantly with gender, baseline WBC, PMN and PLT counts, and nicotine consumption. Dexamethasone and lenograstim administration was more effective than lenograstim administration alone (p < 0.001). Side effects of granulocyte mobilization were generally mild: bone pain in 31.4%, headache in 19.6%, and fatigue in 15.7% of donors. During follow-up (4 weeks), pruritus and/or exanthema were reported in 17.6% of donors. Conclusions Granulocyte mobilization with lenograstim with or without dexamethasone was a safe and effective regimen for granulocyte mobilization. Side effects were tolerable and milder than those seen in peripheral blood stem cell donors. Long-term monitoring of granulocyte donors is important to establish optimal standards for the procedure. PMID:24179474

  6. Robotic-Assisted Laparoscopic Donor Nephrectomy: Decreasing Length of Stay

    PubMed Central

    Cohen, Ari J.; Williams, Darin S.; Bohorquez, Humberto; Bruce, David S.; Carmody, Ian C.; Reichman, Trevor; Loss, George E.

    2015-01-01

    Background The number of robotic operations performed with the da Vinci Surgical System has increased during the past decade. This system allows for greater maneuverability and control than hand-assisted laparoscopic procedures, resulting in less tissue manipulation and irritation. Methods We retrospectively analyzed the results of 100 consecutive robotic-assisted laparoscopic donor nephrectomies and compared them to our most recent 20 hand-assisted laparoscopic donor nephrectomies. Results Between May 2008 and June 2012, 120 laparoscopic donor nephrectomies were performed at Ochsner Clinic Foundation. Of those, 100 live kidney donors underwent robotic-assisted laparoscopic donor nephrectomies. Surgical time and hospital length of stay improved after the first 20 patients receiving robotic-assisted laparoscopic nephrectomies, which was considered the learning curve. Sixty percent of patients who underwent robotic-assisted laparoscopic donor nephrectomies were released on postoperative day 1 compared to 45% of patients who underwent hand-assisted laparoscopic techniques. Conclusion In our experience, robotic-assisted laparoscopic donor nephrectomy resulted in decreased postoperative length of stay that decreased the global cost of the procedure and allowed our institution to admit more patients. PMID:25829876

  7. Serological responses to experimental Norwalk virus infection measured using a quantitative duplex time-resolved fluorescence immunoassay.

    PubMed

    Kavanagh, Owen; Estes, Mary K; Reeck, Amanda; Raju, Ravikiran M; Opekun, Antone R; Gilger, Mark A; Graham, David Y; Atmar, Robert L

    2011-07-01

    A quantitative duplex time-resolved fluorescence assay, dissociation-enhanced lanthanide fluorescent immunoassay (DELFIA), was developed to measure Norwalk virus (NV)-specific IgA and IgG antibodies simultaneously. The duplex assay showed superior performance by detecting seroconversion following experimental NV infection at an earlier time point than a reference total immunoglobulin enzyme-linked immunosorbent assay (ELISA). PMID:21593238

  8. Development of a Luminex Immunoassay for Simultaneous Detection of Shiga toxins and Escherichia coli O157 in Ground Beef

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Shiga toxin-producing Escherichia coli (STEC) are costly foodborne pathogens. As the global prevalence of STEC in beef products ranges from 0.01% to 62.5%, there is a need for developing a rapid screening assay. Here, we developed a Luminex immunoassay to screen for Shiga toxin 1 (Stx1), Shiga toxi...

  9. Strip-based immunoassay for the simultaneous detection of the neonicotinoid insecticides imidacloprid and thiamethoxam in agricultural products

    Technology Transfer Automated Retrieval System (TEKTRAN)

    A semiquantitative strip immunoassay was developed for the rapid detection of imidacloprid and thiamethoxam in agricultural products using specific nanocolloidal gold-labeled monoclonal antibodies. The conjugates of imidacloprid-BSA and thiamethoxam-BSA and goat anti-mouse IgG were coated on the ni...

  10. A solid-phase immunoassay of protease-resistant prion protein with filtration blotting involving sodium dodecyl sulfate.

    PubMed

    Kobayashi, Yoshiteru; Kohno, Naoyuki; Wanibe, Shoko; Hirayasu, Kazunari; Uemori, Hitoshi; Tagawa, Yuichi; Yokoyama, Takashi; Shinagawa, Morikazu

    2006-02-15

    The precise diagnosis for bovine spongiform encephalopathy (BSE) is crucial for preventing new transmission to humans. Several testing procedures are reported for determining protease-resistant prion protein in various tissues as a major hallmark of prion diseases such as BSE, scrapie, and Creutzfeldt-Jakob disease. However, contamination of materials from tissues or degradation of the specimens sometimes disturbs the accuracy of the assay. Here, we have developed a novel method for solid-phase immunoassay of the disease-specific conformational isoform, PrP(Sc), using filtration blotting of protein in the presence of sodium dodecyl sulfate (SDS) followed by a filtration-based immunoassay with a single anti-prion protein antibody, together with the improved fractionation procedure involving high concentrations of surfactant/detergent. The SDS/heat treatment renders unfolded PrP(Sc) quantitative retention on a polyvinylidene difluoride filter and allows enhancement of the analyte signal with immunodetection; thus, all of the tested specimens are determined with 100% accuracy. In addition, the immunoassay is completed in approximately 1h, indicating its usefulness not only for the screening of BSE specimens but probably also for the postmortem BSE diagnosis of fallen stock as the antibody recognizes the core part of PrP(Sc). The solid-phase immunoassay method, including the filtration blotting with SDS, would be applicable to determining even more sensitively proteins other than PrP(Sc), especially those having rigid conformations. PMID:16289444

  11. DEVELOPMENT OF A MAGNETIC PARTICLE IMMUNOASSAY FOR POLYBROMINATED DIPHENYL ETHERS AND APPLICATION TO ENVIRONMENTAL AND FOOD MATRICES.

    Technology Transfer Automated Retrieval System (TEKTRAN)

    A sensitive magnetic particle enzyme-linked immunoassay (ELISA) was developed to analyze polybrominated diphenyl ethers (PBDEs) in water, milk, fish, and soil samples. The assay was rapid and can be used to analyze fifty samples in about one hour after sample cleanup. The assay has a limit of det...

  12. EVALUATION OF IMMUNOASSAY METHODS FOR DETERMINATION OF 3,5,6-TRICHLORO-2-PYRIDINOL IN MULTIPLE SAMPLE MEDIA

    EPA Science Inventory

    Two enzyme-linked immunosorbent assay (ELISA) methods were evaluated for the determination of 3,5,6-trichloro-2-pyridinol (3,5,6-TCP) in multiple sample media (dust, soil, food, and urine). The dust and soil samples were analyzed by the RaPID (TM) commercial immunoassay testing ...

  13. DETECTION OF HEAVY METALS BY IMMUNOASSAY: OPTIMIZATION AND VALIDATION OF A RAPID, PORTABLE ASSAY FOR IONIC CADMIUM (R824029)

    EPA Science Inventory

    An immunoassay is described that measured Cd(II) in aqueous samples at
    concentrations from approximately 7 to 500 ppb. The assay utilized a monoclonal
    antibody that bound tightly to a cadmium-ethylenediaminetetraacetic acid (EDTA)
    complex but not to metal-free EDTA...

  14. Development of a multiplex microsphere immunoassay for the quantitation of salivary antibody responses to selected waterborne pathogens

    EPA Science Inventory

    Saliva has an important advantage over serum as a medium for antibody detection due to non-invasive sampling, which is critical for community-based epidemiological surveys. The development of a Luminex multiplex immunoassay for measurement of salivary IgG and IgA responses to pot...

  15. DEVELOPMENT OF A MAGNETIC PARTICLE ENZYME IMMUNOASSAY AND ITS APPLICATION TO THE MEASUREMENT OF TRICLOSAN IN WATER AND WASTEWATER

    Technology Transfer Automated Retrieval System (TEKTRAN)

    A sensitive magnetic particle-based immunoassay for triclosan (5-chloro-2-(2,4-dichlorophenoxy)phenol) was developed. Rabbit antiserum was produced by immunizing the rabbit with 6-(5-chloro-2-(2,4-dichlorophenoxy)phenoxy)hexanoic acid-keyhole limpet hemocyanin. The triclosan ligand and horse radis...

  16. Application of Rapid Dot Blot Immunoassay for Detection of Salmonella enterica Serovar Enteritidis in Eggs, Poultry, and Other Foods

    PubMed Central

    Yoshimasu, Mark Akira; Zawistowski, Jerzy

    2001-01-01

    Salmonella enterica serovar Enteritidis was detected in artificially inoculated eggs within 24 h through a rapid monoclonal antibody-based dot blot immunoassay. Detection in poultry and other products required 28 h. Samples were directly enriched in homogenized egg without the need for pre- or postenrichment steps. Serovar Enteritidis was detected in the presence of other bacteria when outcompeted 1:400. PMID:11133480

  17. Sensitive immunoassay detection of multiple environmental chemicals on protein microarrays using DNA/dye conjugate as a fluorescent label

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Indirect competitive immunoassays were developed on protein microarrays for the sensitive and simultaneous detection of multiple environmental chemicals in one sample. In this assay, a DNA/SYTOX Orange conjugate was employed as antibody labels to increase the fluorescence signal and sensitivity. Ep...

  18. Application of computer-assisted molecular modeling (CAMM) for immunoassay of low molecular weight food contaminants: A review

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Immunoassay for low molecular weight food contaminants, such as pesticides, veterinary drugs, and mycotoxins is now a well-established technique which meets the demands for a rapid, reliable, and cost-effective analytical method. However, due to limited understanding of the fundamental aspects of i...

  19. Bartonella spp. Bacteremia in Blood Donors from Campinas, Brazil

    PubMed Central

    Pitassi, Luiza Helena Urso; de Paiva Diniz, Pedro Paulo Vissotto; Scorpio, Diana Gerardi; Drummond, Marina Rovani; Lania, Bruno Grosselli; Barjas-Castro, Maria Lourdes; Gilioli, Rovilson; Colombo, Silvia; Sowy, Stanley; Breitschwerdt, Edward B.; Nicholson, William L.; Velho, Paulo Eduardo Neves Ferreira

    2015-01-01

    Bartonella species are blood-borne, re-emerging organisms, capable of causing prolonged infection with diverse disease manifestations, from asymptomatic bacteremia to chronic debilitating disease and death. This pathogen can survive for over a month in stored blood. However, its prevalence among blood donors is unknown, and screening of blood supplies for this pathogen is not routinely performed. We investigated Bartonella spp. prevalence in 500 blood donors from Campinas, Brazil, based on a cross-sectional design. Blood samples were inoculated into an enrichment liquid growth medium and sub-inoculated onto blood agar. Liquid culture samples and Gram-negative isolates were tested using a genus specific ITS PCR with amplicons sequenced for species identification. Bartonella henselae and Bartonella quintana antibodies were assayed by indirect immunofluorescence. B. henselae was isolated from six donors (1.2%). Sixteen donors (3.2%) were Bartonella-PCR positive after culture in liquid or on solid media, with 15 donors infected with B. henselae and one donor infected with Bartonella clarridgeiae. Antibodies against B. henselae or B. quintana were found in 16% and 32% of 500 blood donors, respectively. Serology was not associated with infection, with only three of 16 Bartonella-infected subjects seropositive for B. henselae or B. quintana. Bartonella DNA was present in the bloodstream of approximately one out of 30 donors from a major blood bank in South America. Negative serology does not rule out Bartonella spp. infection in healthy subjects. Using a combination of liquid and solid cultures, PCR, and DNA sequencing, this study documents for the first time that Bartonella spp. bacteremia occurs in asymptomatic blood donors. Our findings support further evaluation of Bartonella spp. transmission which can occur through blood transfusions. PMID:25590435

  20. Ethical issues relating to renal transplantation from prediabetic living donor

    PubMed Central

    2014-01-01

    Background In Mexico, diabetes mellitus is the main cause of end???stage kidney disease, and some patients may be transplant candidates. Organ supply is limited because of cultural issues. And, there is a lack of standardized clinical guidelines regarding organ donation. These issues highlight the tension surrounding the fact that living donors are being selected despite being prediabetic. This article presents, examines and discusses using the principles of non-maleficience, autonomy, justice and the constitutionally guaranteed right to health, the ethical considerations that arise from considering a prediabetic person as a potential kidney donor. Discussion Diabetes is an absolute contraindication for donating a kidney. However, the transplant protocols most frequently used in Mexico do not consider prediabetes as exclusion criteria. In prediabetic persons there are well known metabolic alterations that may compromise the long???term outcomes of the transplant if such donors are accepted. Even so, many of them are finally included because there are not enough donor candidates. Both, families and hospitals face the need to rapidly accept prediabetic donors before the clinical conditions of the recipient and the evolution of the disease exclude him/her as a transplant candidate; however, when using a kidney potentially damaged by prediabetes, neither the donor抯 nor the recipient抯 long term health is usually considered. Considering the ethical implication as well as the clinical and epidemiological evidence, we conclude that prediabetic persons are not suitable candidates for kidney donation. This recommendation should be taken into consideration by Mexican health institutions who should rewrite their transplant protocols. Summary We argue that the decision to use a kidney from a living donor known to be pre-diabetic or from those persons with family history of T2DM, obesity, hypertension, or renal failure, should be considered unethical in Mexico if the donor bases the decision to donate on socially acceptable norms rather than informed consent as understood in modern medicine. PMID:24935278

  1. Shallow-donor lasers in uniaxially stressed silicon

    SciTech Connect

    Kovalevsky, K. A. Zhukavin, R. Kh.; Tsyplenkov, V. V.; Shastin, V. N.; Abrosimov, N. V.; Riemann, H.; Pavlov, S. G.; Huebers, H.-W.

    2013-02-15

    The effects of the terahertz-stimulated emission of Group-V donors (phosphorus, antimony, arsenic, bismuth) in uniaxially stressed silicon, excited by CO{sub 2} laser radiation are experimentally studied. It is shown that uniaxial compressive stress of the crystal along the [100] direction increases the gain and efficiency of stimulated radiation, significantly decreasing the threshold pump intensity. The donor frequencies are measured and active transitions are identified in stressed silicon. The dependence of the residual population of active donor states on the uniaxial compressive stress along the [100] direction is theoretically estimated.

  2. Recruiting gamete donors: response to Craft and Thornhill.

    PubMed

    Daniels, Ken

    2005-04-01

    This response challenges the notion that paying more money, in whatever form, will overcome the anxieties of gamete donors regarding the removal of anonymity. An overview of the literature on payment and donor recruitment indicates that this is a complex issue. That literature does provide 'pointers' to the types of persons who should be 'targeted' in an information sharing and non-payment system. The experience in Sweden, both in relation to donor recruitment and parents' information-sharing with their children, is clarified, as the evidence is not always presented accurately. PMID:15901446

  3. Hydrogen-accelerated thermal donor formation in Czochralski silicon

    SciTech Connect

    Stein, H.J. ); Hahn, S.K. )

    1990-01-01

    Acceleration of thermal donor formation at 400 {degree}C in Czochralski Si by a hydrogen plasma has been observed using low-temperature infrared absorption and spreading resistance probe measurements. The accelerated formation in as-grown Si is attributed to hydrogen diffusion and catalyzed conversion of electrically inactive nuclei to thermal donors. When the nuclei concentration is small such as in rapid thermal annealed Si, hydrogen interaction at SiO bonds is suggested as the rate-limiting step in thermal donor formation.

  4. Whole genome comparison of donor and cloned dogs

    PubMed Central

    Kim, Hak-Min; Cho, Yun Sung; Kim, Hyunmin; Jho, Sungwoong; Son, Bongjun; Choi, Joung Yoon; Kim, Sangsoo; Lee, Byeong Chun; Bhak, Jong; Jang, Goo

    2013-01-01

    Cloning is a process that produces genetically identical organisms. However, the genomic degree of genetic resemblance in clones needs to be determined. In this report, the genomes of a cloned dog and its donor were compared. Compared with a human monozygotic twin, the genome of the cloned dog showed little difference from the genome of the nuclear donor dog in terms of single nucleotide variations, chromosomal instability, and telomere lengths. These findings suggest that cloning by somatic cell nuclear transfer produced an almost identical genome. The whole genome sequence data of donor and cloned dogs can provide a resource for further investigations on epigenetic contributions in phenotypic differences. PMID:24141358

  5. Vasectomy as a reason for donor insemination.

    PubMed

    Humphrey, M; Humphrey, H

    1993-07-01

    Among 200 couples referred for counselling with a view to donor insemination (DI) were 21 whose request arose from an earlier vasectomy. This had been judged irreversible, apart from one case where the man had been sterilised for genetic reasons and therefore had not sought reversal. An overall incidence of 10.5% indicates that vasectomy may lead to fertility problems in the event of remarriage, which has become more common with an increasing divorce rate. All 21 husbands but only 3 wives had been previously married; 3 former wives had died, otherwise all previous marriages had ended in divorce. Most husbands were considerably older than their second wives, especially as compared with the majority of men in the larger series who were still in a first marriage. Their relative maturity may have helped them to respond more sensitively to their wives' maternal needs when they already had children of their own. During a follow-up interval of up to 8 years, 2 couples had separated after a brief relationship but 9 of the other 12 contacted had become parents by DI. The 3 remaining couples included a wife who at the latest report was 5 months pregnant, and 2 who were still pursuing treatment. Despite the low failure rate in this small series we should bear in mind that DI is not an ideal solution to the problem of male infertility, nor always even acceptable.(ABSTRACT TRUNCATED AT 250 WORDS) PMID:8351539

  6. Antibodies to HLA-E may account for the non-donor-specific anti-HLA class-Ia antibodies in renal and liver transplant recipients.

    PubMed

    Ravindranath, Mepur H; Pham, Tho; Ozawa, Miyuki; Terasaki, Paul I

    2012-01-01

    The non-donor-specific anti-HLA-Ia antibodies correlate significantly with lower graft survival in organ transplant patients. Based on our earlier findings that anti-HLA-E murine monoclonal antibodies (MEM-E/02 and 3D12) reacted with different HLA-Ia alleles and the peptides shared by HLA-E and HLA class, Ia alleles inhibited the HLA-Ia reactivity of the anti-HLA-E antibodies in normal non-alloimmunized males, the possibility of that anti-HLA-E IgG may account for the non-donor-specific anti-HLA-Ia antibodies in the allograft recipients was examined by multiplex-Luminex-immunoassay. About 73% of renal and 53% of liver transplant patients' sera with high level of anti-HLA-E IgG showed reactivity to different non-donor HLA-Ia alleles. About 50% renal and 52% liver allograft recipients' sera with low level of anti-HLA-E IgG had no reactivity to any HLA-Ia alleles; however, the IgG isolated from the same sera with protein-G columns showed the presence of anti-HLA-E IgG with HLA-Ia reactivity. Furthermore, both recombinant HLA-E and the IgG-free serum containing soluble HLA-E (sHLA-E) inhibited HLA-Ia reactivity of anti-HLA-E murine monoclonal IgG significantly. The data suggest that the HLA-Ia reactivity of the anti-HLA-E antibody accounts for the non-donor-specific anti-HLA-Ia antibodies. It is proposed that the sHLA-E heavy chain, shed in circulation after organ transplantation, may expose cryptic epitopes of HLA-E to elicit anti-HLA-E IgG antibodies, which may cross react with HLA-Ia alleles due to the peptide sequences shared between them. This study provides a new explanation for the presence of non-donor-specific antibodies for non-existing HLA-Ia alleles, frequently observed and correlated with survival in organ transplant recipients. PMID:22190577

  7. Donor Funding for Newborn Survival: An Analysis of Donor-Reported Data, 20022010

    PubMed Central

    Pitt, Catherine; Lawn, Joy E.; Ranganathan, Meghna; Mills, Anne; Hanson, Kara

    2012-01-01

    Background Neonatal mortality accounts for 43% of global under-five deaths and is decreasing more slowly than maternal or child mortality. Donor funding has increased for maternal, newborn, and child health (MNCH), but no analysis to date has disaggregated aid for newborns. We evaluated if and how aid flows for newborn care can be tracked, examined changes in the last decade, and considered methodological implications for tracking funding for specific population groups or diseases. Methods and Findings We critically reviewed and categorised previous analyses of aid to specific populations, diseases, or types of activities. We then developed and refined key terms related to newborn survival in seven languages and searched titles and descriptions of donor disbursement records in the Organisation for Economic Co-operation and Development's Creditor Reporting System database, 20022010. We compared results with the Countdown to 2015 database of aid for MNCH (20032008) and the search strategy used by the Institute for Health Metrics and Evaluation. Prior to 2005, key terms related to newborns were rare in disbursement records but their frequency increased markedly thereafter. Only two mentions were found of 搒tillbirth and only nine references were found to 揻etus in any spelling variant or language. The total value of non-research disbursements mentioning any newborn search terms rose from US$38.4 million in 2002 to US$717.1 million in 2010 (constant 2010 US$). The value of non-research projects exclusively benefitting newborns fluctuated somewhat but remained low, at US$5.7 million in 2010. The United States and the United Nations Children's Fund (UNICEF) provided the largest value of non-research funding mentioning and exclusively benefitting newborns, respectively. Conclusions Donor attention to newborn survival has increased since 2002, but it appears unlikely that donor aid is commensurate with the 3.0 million newborn deaths and 2.7 million stillbirths each year. We recommend that those tracking funding for other specific population groups, diseases, or activities consider a key term search approach in the Creditor Reporting System along with a detailed review of their data, but that they develop their search terms and interpretations carefully, taking into account the limitations described. Please see later in the article for the Editors' Summary PMID:23118619

  8. Pressure dependence of donor excitation spectra in AlSb

    SciTech Connect

    Hsu, L.; McCluskey, M.D.; Haller, E.E.

    2002-01-16

    We have investigated the behavior of ground to bound excited-state electronic transitions of Se and Te donors in AlSb as a function of hydrostatic pressure. Using broadband far-infrared Fourier transform spectroscopy, we observe qualitatively different behaviors of the electronic transition energies of the two donors. While the pressure derivative of the Te transition energy is small and constant, as might be expected for a shallow donor, the pressure derivatives of the Se transition energies are quadratic and large at low pressures, indicating that Se is actually a deep donor. In addition, at pressures between 30 and 50 kbar, we observe evidence of an anti-crossing between one of the selenium electronic transitions and a two-phonon mode.

  9. 21 CFR 640.51 - Suitability of donors.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ...Plasmapheresis donors shall meet the criteria for suitability prescribed in 640.63, excluding the phrase 搊ther than malaria in paragraph (c) (9) of that section. Informed consent shall be required as prescribed in 640.61. [42 FR...

  10. 21 CFR 640.51 - Suitability of donors.

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ...Plasmapheresis donors shall meet the criteria for suitability prescribed in 640.63, excluding the phrase 搊ther than malaria in paragraph (c) (9) of that section. Informed consent shall be required as prescribed in 640.61. [42 FR...

  11. 21 CFR 640.51 - Suitability of donors.

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ...Plasmapheresis donors shall meet the criteria for suitability prescribed in 640.63, excluding the phrase 搊ther than malaria in paragraph (c) (9) of that section. Informed consent shall be required as prescribed in 640.61. [42 FR...

  12. 21 CFR 640.51 - Suitability of donors.

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ...Plasmapheresis donors shall meet the criteria for suitability prescribed in 640.63, excluding the phrase 搊ther than malaria in paragraph (c) (9) of that section. Informed consent shall be required as prescribed in 640.61. [42 FR...

  13. 21 CFR 640.51 - Suitability of donors.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ...Plasmapheresis donors shall meet the criteria for suitability prescribed in 640.63, excluding the phrase 搊ther than malaria in paragraph (c) (9) of that section. Informed consent shall be required as prescribed in 640.61. [42 FR...

  14. Donor characterization in ZnO by thermally stimulated luminescence

    SciTech Connect

    Ji, jianfeng; Boatner, Lynn A; Selim, F. A.

    2014-01-01

    Low temperature thermo-luminescence (TL) was applied to measurements of the ionization energy of donors in ZnO. Three hydrogen-related donors were characterized with ionization energies of 36, 47 and 55 meV - values that are in complete agreement with previous reports. The donor types can be switched by relevant thermal treatments. These measurements also revealed the presence of two distinct sources for the green luminescence in ZnO. This work indicates that TL can be used to measure the donor energies in luminescent semiconductors. This approach can be particularly useful for thin-film investigations when the results of Hall-effect measurements are obscured by contributions from conductive interfaces or substrates.

  15. Errors and Omissions: Donor Compensation Policies and Richard Titmuss.

    PubMed

    Penrod, Joshua; Farrugia, Albert

    2015-12-01

    Many global and national systems of regulation of blood donors and donor compensation rely for intellectual support on Richard Titmuss's views, represented in The Gift Relationship. Based on selective interpretation of data from the 1960s, Titmuss engineered an ethical view pertaining to donors and, in so doing, created not only ongoing stereotypes, but created a cause for followers to perpetuate misunderstandings about the nature of such donations. In many cases, donors are, in fact compensated, but regulatory systems persevere in using definitional fig leaves in order to perpetuate an ongoing political goal of diminishing private sector participation in health care. However, in more recent works, including new views of critical sociology and evolutionary psychology, the Titmuss worldview has been turned upside-down. Evidence readily available today proves the safety of compensated donation and the lives saved by encouraging policies for both compensated and non-compensated donation. PMID:25680344

  16. Retrovirus Epidemiology Donor Study-II (REDS-II)

    ClinicalTrials.gov

    2015-11-09

    Acquired Immunodeficiency Syndrome; Blood Donors; Blood Transfusion; HIV Infections; HIV-1; HIV-2; HTLV-I; HTLV-II; Retroviridae Infections; Hepatitis, Viral, Human; Hepatitis B; Hepacivirus; West Nile Virus

  17. High precision quantum control of single donor spins in silicon

    E-print Network

    Rajib Rahman; Cameron J. Wellard; Forrest R. Bradbury; Marta Prada; Jared H. Cole; Gerhard Klimeck; Lloyd C. L. Hollenberg

    2007-05-15

    The Stark shift of the hyperfine coupling constant is investigated for a P donor in Si far below the ionization regime in the presence of interfaces using Tight-binding and Band Minima Basis approaches and compared to the recent precision measurements. The TB electronic structure calculations included over 3 million atoms. In contrast to previous effective mass based results, the quadratic Stark coefficient obtained from both theories agrees closely with the experiments. This work represents the most sensitive and precise comparison between theory and experiment for single donor spin control. It is also shown that there is a significant linear Stark effect for an impurity near the interface, whereas, far from the interface, the quadratic Stark effect dominates. Such precise control of single donor spin states is required particularly in quantum computing applications of single donor electronics, which forms the driving motivation of this work.

  18. Improvement of an enzyme immunoassay for the determination of mercury (II)

    SciTech Connect

    Marx, A.; Kroetz, E.; Hock, B.

    1998-07-01

    Three systems were tested for the optimization of a heterogeneous noncompetitive enzyme immunoassay (EIA) for the determination of Hg (II). The sensitivity of the nonoptimized Hg-EIA with a detection limit of 2.1 {micro}g/L Hg (II) was improved by an avidin-biotin-complex (ABC) amplification system to a 2-fold lower detection limit (1.1 {micro}g/L Hg (II)). A conventional competitive EIA with the competition reaction between bound and free Hg (II) for antibody (ab) binding sites yielded a detection limit of 1.0 {micro}g/L Hg (II). Further improvement of sensitivity could be achieved by a competitive displacement EIA. In this case ab molecules bound to immobilized haptens are displaced in the next step by free Hg (II). The detection limit of the displacement approach is 0.4 {micro}g/L Hg (II).

  19. Noninvasive analysis of fecal reproductive hormone metabolites in female veiled chameleons (Chamaeleo calyptratus) by enzyme immunoassay.

    PubMed

    Kummrow, Maya S; Gilman, Christine; Mackie, Paula; Smith, Dale A; Mastromonaco, Gabriela F

    2011-01-01

    The noninvasive technique of gonadal steroid metabolite measurement in feces for evaluation of reproductive activity has proven an effective and important tool for population management in various captive species, but has not yet been validated and used in reptile species. In this study, enzyme immunoassays (EIAs) were validated for the analysis of fecal samples from female veiled chameleons (Chamaeleo calyptratus) for estrogen (E2), testosterone (T), and progesterone (P) and their metabolites. High performance liquid chromatography and physiological methods (GnRH stimulation) were used for the validation of the assays. Biological events, such as skin color changes indicative of ovarian activity and oviposition, correlated with the cyclical pattern of E2, T and P metabolites in feces over a period of two reproductive cycles. This is the first study to report frequent longitudinal measurements of fecal hormone levels by EIA in a reptile species. PMID:21319212

  20. Rapid Simultaneous Detection of Anti-protozoan Drugs Using a Lateral-Flow Immunoassay Format.

    PubMed

    Fitzgerald, Jenny; Leonard, Paul; Danaher, Martin; O'Kennedy, Richard

    2015-05-01

    This research describes the development of a multi-analyte lateral-flow immunoassay intended for the simultaneous detection of three anti-protozoan drugs (coccidiostats). These drugs, namely, halofuginone, toltrazuril and diclazuril, are used in the treatment of Eimeria spp. infections in cattle, pigs, chickens and turkeys. Coloured carboxylated microspheres were coated with each of the detection antibodies and employed in a lateral-flow assay format for detection of these residues in eggs. Using this approach, halofuginone was detectable at a limit of 10 ng/mL or greater, toltrazuril at 100 ng/mL and, similarly, diclazuril had a detection limit of 100 ng/mL, which is below the maximum allowed residue limit for all three as outlined by EU regulation. This simple cost-efficient assay and analysis method could pave the way for more efficient simultaneous monitoring of small-molecule residues in the future. PMID:25832180