Science.gov

Sample records for disc susceptibility testing

  1. 21 CFR 866.1620 - Antimicrobial susceptibility test disc.

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ... 21 Food and Drugs 8 2013-04-01 2013-04-01 false Antimicrobial susceptibility test disc. 866.1620 Section 866.1620 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) MEDICAL DEVICES IMMUNOLOGY AND MICROBIOLOGY DEVICES Diagnostic Devices § 866.1620...

  2. 21 CFR 866.1620 - Antimicrobial susceptibility test disc.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... 21 Food and Drugs 8 2010-04-01 2010-04-01 false Antimicrobial susceptibility test disc. 866.1620 Section 866.1620 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) MEDICAL DEVICES IMMUNOLOGY AND MICROBIOLOGY DEVICES Diagnostic Devices § 866.1620...

  3. 21 CFR 866.1620 - Antimicrobial susceptibility test disc.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... 21 Food and Drugs 8 2011-04-01 2011-04-01 false Antimicrobial susceptibility test disc. 866.1620 Section 866.1620 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) MEDICAL DEVICES IMMUNOLOGY AND MICROBIOLOGY DEVICES Diagnostic Devices § 866.1620...

  4. 21 CFR 866.1620 - Antimicrobial susceptibility test disc.

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ... 21 Food and Drugs 8 2014-04-01 2014-04-01 false Antimicrobial susceptibility test disc. 866.1620 Section 866.1620 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) MEDICAL DEVICES IMMUNOLOGY AND MICROBIOLOGY DEVICES Diagnostic Devices § 866.1620...

  5. 21 CFR 866.1620 - Antimicrobial susceptibility test disc.

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ... 21 Food and Drugs 8 2012-04-01 2012-04-01 false Antimicrobial susceptibility test disc. 866.1620 Section 866.1620 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) MEDICAL DEVICES IMMUNOLOGY AND MICROBIOLOGY DEVICES Diagnostic Devices § 866.1620...

  6. BSAC standardized disc susceptibility testing method (version 8).

    PubMed

    Andrews, J M

    2009-09-01

    azithromycin, clarithromycin, erythromycin and telithromycin (Table 15); clarithromycin and erythromycin MIC and zone diameter BPs for Moraxella catarrhalis (Table 16); azithromycin MIC BPs for Neisseria gonorrhoeae (Table 17); chloramphenicol and rifampicin MIC BPs for Neisseria meningitidis (Table 18); azithromycin MIC BPs for Haemophilus influenzae (Table 19); MIC BPs for metronidazole for Bacteroides fragilis, Bacteroides thetaiotaomicron and Clostridium perfringens (Tables 23-25, respectively); susceptibility testing of Listeria spp. (Appendix 3); the acceptable range for ATCC 25923 to a 10 microg tobramycin disc (Table 26). PMID:19587067

  7. Rifaximin disc diffusion test for in vitro susceptibility testing of Clostridium difficile

    PubMed Central

    Huhulescu, Steliana; Sagel, Ulrich; Fiedler, Anita; Pecavar, Verena; Blaschitz, Marion; Wewalka, Guenther; Allerberger, Franz

    2011-01-01

    Rifaximin is a rifampicin derivative, poorly absorbed by the gastro-intestinal tract. We studied the in vitro susceptibility to rifamixin of 1082 Clostridium difficile isolates; among these,184 isolates from a strain collection were tested by an in-house rifaximin disc (40 µg) diffusion test, by an in-house rifaximin broth microdilution test, by rifampicin Etest and by rpoB gene sequencing. In the absence of respective CLSI or EUCAST MIC breakpoints for rifaximin and rifampicin against C. difficile we chose MIC ≥32 µg ml−1 as criterion for reduced in vitro susceptibility. To further validate the disc diffusion test 898 consecutive clinical isolates were analysed using the disc diffusion test, the Etest and rpoB gene sequence analysis for all resistant strains. Rifaximin broth microdilution tests of the 184 reference strains yielded rifaximin MICs ranging from 0.001 (n = 1) to ≥1024 µg ml−1 (n = 61); 62 isolates showed a reduced susceptibility (MIC ≥32 µg ml−1). All of these 62 strains showed rpoB gene mutations producing amino acid substitutions; the rifampicin- and rifaximin-susceptible strains showed either a wild-type sequence or silent amino acid substitutions (19 strains). For 11 arbitrarily chosen isolates with rifaximin MICs of >1024 µg ml−1, rifaximin end-point MICs were determined by broth dilution: 4096 µg ml−1 (n = 2), 8192 µg ml−1 (n = 6), 16 384 µg ml−1 (n = 2) and 32 678 µg ml−1 (n = 1). Rifampicin Etests on the 184 C. difficile reference strains yielded MICs ranging from ≤0.002 (n = 117) to ≥32 µg ml−1 (n = 59). Using a 38 mm inhibition zone as breakpoint for reduced susceptibility the use of rifaximin disc diffusion yielded 59 results correlating with those obtained by use of rifaximin broth microdilution in 98.4 % of the 184 strains tested. Rifampicin Etests performed on the 898 clinical isolates revealed that 67 isolates had MICs of ≥32 µg ml−1. There were no

  8. Rifaximin disc diffusion test for in vitro susceptibility testing of Clostridium difficile.

    PubMed

    Huhulescu, Steliana; Sagel, Ulrich; Fiedler, Anita; Pecavar, Verena; Blaschitz, Marion; Wewalka, Guenther; Allerberger, Franz; Indra, Alexander

    2011-08-01

    Rifaximin is a rifampicin derivative, poorly absorbed by the gastro-intestinal tract. We studied the in vitro susceptibility to rifamixin of 1082 Clostridium difficile isolates; among these, 184 isolates from a strain collection were tested by an in-house rifaximin disc (40 µg) diffusion test, by an in-house rifaximin broth microdilution test, by rifampicin Etest and by rpoB gene sequencing. In the absence of respective CLSI or EUCAST MIC breakpoints for rifaximin and rifampicin against C. difficile we chose MIC ≥32 µg ml(-1) as criterion for reduced in vitro susceptibility. To further validate the disc diffusion test 898 consecutive clinical isolates were analysed using the disc diffusion test, the Etest and rpoB gene sequence analysis for all resistant strains. Rifaximin broth microdilution tests of the 184 reference strains yielded rifaximin MICs ranging from 0.001 (n = 1) to ≥1024 µg ml(-1) (n = 61); 62 isolates showed a reduced susceptibility (MIC ≥32 µg ml(-1)). All of these 62 strains showed rpoB gene mutations producing amino acid substitutions; the rifampicin- and rifaximin-susceptible strains showed either a wild-type sequence or silent amino acid substitutions (19 strains). For 11 arbitrarily chosen isolates with rifaximin MICs of >1024 µg ml(-1), rifaximin end-point MICs were determined by broth dilution: 4096 µg ml(-1) (n = 2), 8192 µg ml(-1) (n = 6), 16,384 µg ml(-1) (n = 2) and 32,678 µg ml(-1) (n = 1). Rifampicin Etests on the 184 C. difficile reference strains yielded MICs ranging from ≤0.002 (n = 117) to ≥32 µg ml(-1) (n = 59). Using a 38 mm inhibition zone as breakpoint for reduced susceptibility the use of rifaximin disc diffusion yielded 59 results correlating with those obtained by use of rifaximin broth microdilution in 98.4 % of the 184 strains tested. Rifampicin Etests performed on the 898 clinical isolates revealed that 67 isolates had MICs of ≥32 µg ml(-1). There were no discordant

  9. Susceptibility Testing

    MedlinePlus

    ... page helpful? Also known as: Sensitivity Testing; Drug Resistance Testing; Culture and Sensitivity; C & S; Antimicrobial Susceptibility Formal name: Bacterial and Fungal Susceptibility Testing Related tests: Urine Culture ; ...

  10. Comparison of disc diffusion, Etest and broth microdilution for testing susceptibility of carbapenem-resistant P. aeruginosa to polymyxins

    PubMed Central

    van der Heijden, Inneke M; Levin, Anna S; De Pedri, Ewerton H; Fung, Liang; Rossi, Flavia; Duboc, Gisele; Barone, Antonio A; Costa, Silvia F

    2007-01-01

    Background Considering the increasing use of polymyxins to treat infections due to multidrug resistant Gram-negative in many countries, it is important to evaluate different susceptibility testing methods to this class of antibiotic. Methods Susceptibility of 109 carbapenem-resistant P. aeruginosa to polymyxins was tested comparing broth microdilution (reference method), disc diffusion, and Etest using the new interpretative breakpoints of Clinical and Laboratory Standards Institute. Results Twenty-nine percent of isolates belonged to endemic clone and thus, these strains were excluded of analysis. Among 78 strains evaluated, only one isolate was resistant to polymyxin B by the reference method (MIC: 8.0 μg/mL). Very major and major error rates of 1.2% and 11.5% were detected comparing polymyxin B disc diffusion with the broth microdilution (reference method). Agreement within 1 twofold dilution between Etest and the broth microdilution were 33% for polymyxin B and 79.5% for colistin. One major error and 48.7% minor errors were found comparing polymyxin B Etest with broth microdilution and only 6.4% minor errors with colistin. The concordance between Etest and the broth microdilution (reference method) was respectively 100% for colistin and 90% for polymyxin B. Conclusion Resistance to polymyxins seems to be rare among hospital carbapenem-resistant P. aeruginosa isolates over a six-year period. Our results showed, using the new CLSI criteria, that the disc diffusion susceptibility does not report major errors (false-resistant results) for colistin. On the other hand, showed a high frequency of minor errors and 1 very major error for polymyxin B. Etest presented better results for colistin than polymyxin B. Until these results are reproduced with a large number of polymyxins-resistant P. aeruginosa isolates, susceptibility to polymyxins should be confirmed by a reference method. PMID:17697363

  11. Comparative assessment of CDS, CLSI disc diffusion and Etest techniques for antimicrobial susceptibility testing of Neisseria gonorrhoeae: a 6-year study

    PubMed Central

    Singh, Vikram; Kakran, Monika; Ramesh, V

    2012-01-01

    Background A variety of techniques are available for antimicrobial susceptibility testing of Neisseria gonorrhoeae. Objective The aim of this study was to find a cost-effective, reliable and easily applicable microbiological method to detect antimicrobial susceptibilities of N. gonorrhoeae in resource-poor countries. Design Prospective study. Setting Male and female STD clinic of Regional STD Teaching, Training and Research Centre, New Delhi, India. Participants N. gonorrhoeae isolates from all male and female patients presenting with acute gonococcal urethritis and cervical discharge. Material and methods A total of 295 consecutive N. gonorrhoeae isolates during 2005–2010 was used to compare the Clinical and Laboratory Standards Institute (CLSI) and CDS disc diffusion technique with Etest by performing antimicrobial susceptibility testing in parallel for penicillin, tetracycline, ceftriaxone, ciprofloxacin and spectinomycin. WHO reference strains were used as controls. Results CDS disc diffusion zones of inhibition showed that complete percentage agreement for penicillin, ciprofloxacin and tetracycline was high with their analogous Etest minimal inhibitory concentrations in comparison to CLSI disc diffusion technique, that is, 91.5%, 92.9% and 99.3% versus 87.5%, 88.5% and 74.9%, respectively. CDS results had less number of major and minor category discrepancies in comparison to CLSI and CDS method showed excellent correlation coefficient (r=1) with Etest for all five antimicrobial agents tested in comparison to CLSI (r=0.92). It was very poor (r=0.61) by CLSI method for tetracycline. The correlation coefficients between the two methods and the Etest were identical if tetracycline was removed from the CLSI analysis. Conclusions The CDS technique is an attractive alternative for N. gonorrhoeae susceptibility testing and is recommended for monitoring the antimicrobial susceptibility in less developed and resource-poor settings to facilitate enhanced antimicrobial

  12. Comparative Analysis of Disc Diffusion and E-test with Broth Micro-dilution for Susceptibility Testing of Clinical Candida Isolates Against Amphotericin B, Fluconazole, Voriconazole and Caspofungin

    PubMed Central

    Bhattacharyya, Sayan; Gupta, Prashant; Banerjee, Gopa; Singh, Mastan

    2015-01-01

    Background Antifungal susceptibility testing remains an area of intense interest because of the increasing number of clinical isolates resistant to antifungal therapy. Clinical and Laboratory Standards Institute has proposed reference broth micro dilution (BMD) method for susceptibility testing. The reference method is time-consuming and poorly suited for the routine clinical laboratory setting. Agar-based susceptibility testing methods, disk diffusion (DD) method and the E-test method can be an easier, reliable and less time consuming alternative for the BMD method. Aim To compare the results of Amphotericin B, fluconazole, voriconazole, and Caspofungin susceptibility testing by DD, and the E-test method with the CLSI reference method for clinical Candida isolates. Materials and Methods Broth Microdilution (BMD), E-test and Disk diffusion testing of the various clinical Candida isolates was performed in accordance with CLSI documents. The results obtained were analysed and compared. Results The categorical agreement for Amphotericin B, fluconazole, voriconazole, and Caspofungin susceptibility results by E-test and DD method was 65.2%, 67.4%; 100%, 82.6%; 100%, 100%; 100%, 97.8% respectively. Conclusion The agar-based E-test and disk diffusion methods are reliable alternatives to the BMD method for Candida isolates when test susceptible to fluconazole, voriconazole, and Caspofungin, however the susceptibility testing results must be interpreted with caution in case of Amphotericin B. PMID:26675415

  13. Evaluation of the mastascanelite image analysis system for measuring zones of inhibition in disc diffusion susceptibility tests.

    PubMed

    Clarke, R E H; Winstanley, T G; Ridgway, E J

    2003-01-01

    In this evaluation a mastascanelite image analysis system is compared with manual measurement of disc diffusion inhibition zones. Data for 213 diverse organisms and a total of 1679 organism/antibiotic combinations gave an overall correlation coefficient of 0.988. The mean difference between readings was +0.425 mm, with 95% confidence limits of +/-2.94 mm, and the majority of scanned zones (97.51%) fell within +/-3 mm of the manual measurement. The mastascanelite system forms part of a laboratory suite and was found to be objective, accurate and rapid, reading and interpreting each plate in less than a second. Interfacing to the laboratory computer system facilitated data handling and performance control. PMID:14560787

  14. Real-time video imaging as a new and rapid tool for antibiotic susceptibility testing by the disc diffusion method: a paradigm for evaluating resistance to imipenem and identifying extended-spectrum β-lactamases.

    PubMed

    Le Page, Stéphanie; Raoult, Didier; Rolain, Jean-Marc

    2015-01-01

    The disc diffusion method has long been considered the standard technique for antibiotic susceptibility testing (AST) in clinical microbiology laboratories because of its simplicity, reproducibility and low cost compared with commercial automated microdilution systems that are usually more rapid but less sensitive for detecting important mechanisms of resistance. Here we measured reading zone diameters around antibiotics in a series of 25 well-characterised Gram-negative bacteria by the disc diffusion technique in real-time using an Advencis Bio-System instrument consisting of a real-time high-resolution video imager in a dedicated incubator. The susceptibility of wild-type Gram-negative bacteria to imipenem, determined by reading the diameter of inhibition, was detectable as early as 3.5h (mean time 3.7 ± 0.45 h), whereas carbapenemase-producing Gram-negative bacteria could be correctly categorised as early as 3h (mean time 4.2 ± 0.8 h) of incubation. Similarly, the characteristic champagne cork aspect of extended-spectrum β-lactamase (ESBL) could be detected by the system as early as 3.5 h. Moreover, we present here for the first time video movies of the appearance of the diameter of inhibition by disc diffusion in real-time. This preliminary study using a new and innovative technology provides for a renewed interest for microbiologists who wish to continue to use the disc diffusion method as a reference method for AST. New video imaging technology presents a proof of concept that could improve the real-time management of patients with AST within a very rapid turnaround time and can provide a large financial saving for hospitals. PMID:25455851

  15. Comparison of tablets and paper discs for antibiotic sensitivity testing.

    PubMed Central

    Brown, D F; Kothari, D

    1975-01-01

    The value of tablets and paper discs as reservoirs of antimicrobial agents for use in sensitivity testing was compared. Antibiotics that were unstable in paper discs showed no demonstrable loss of activity in tablets over a period of 50 days under adverse storage conditions. The antibiotic content of commercially prepared tablets is very high in comparison with the accepted content of paper discs used in Britain, but not all of the agent is released from tablets during tests. Comparison of the size of zones of inhibition around tablets and standard paper discs indicated that the amount of the various agents released from the tablets varied between 2-6% and 69% of the stated content. In tests of the sensitivity of a range of common pathogenic organisms, the results obtained with the tablet method--when interpreted as recommended by the manufacturer--were generally similar to those obtained with a paper disc method commonly used in British laboratories. In 47% of tests with aminoglycoside antibiotics, however, strains sensitive by the disc method were 'intermediate' or resistant by the tablet method. As with paper discs, it was necessary to press the tablets on to the medium. With adjustment of the 'effective antibiotic content of tablets to bring it into line with the accepted content in paper discs, the stability of antibiotics in the tablets might make them an acceptable alternative to paper discs. PMID:1206124

  16. Stress test for the intervertebra disc joint core

    NASA Astrophysics Data System (ADS)

    Yang, Guobiao; Fang, Ruhua

    2002-05-01

    The inter-vertebra disc is an important part of human lumbar spine, it is valuable to examine the lumbar spine and inter- vertebra disc by means of bio-mechanics. A new testing method is described in this paper for inner stress measuring, the test data is verified by the other test method, which is satisfying, which is satisfying. The results of this paper establish the foundation of the inter-vertebra disc core study, and it is helpful in the clinic and practical application.

  17. Testing hydrodynamics schemes in galaxy disc simulations

    NASA Astrophysics Data System (ADS)

    Few, C. G.; Dobbs, C.; Pettitt, A.; Konstandin, L.

    2016-08-01

    We examine how three fundamentally different numerical hydrodynamics codes follow the evolution of an isothermal galactic disc with an external spiral potential. We compare an adaptive mesh refinement code (RAMSES), a smoothed particle hydrodynamics code (SPHNG), and a volume-discretized mesh-less code (GIZMO). Using standard refinement criteria, we find that RAMSES produces a disc that is less vertically concentrated and does not reach such high densities as the SPHNG or GIZMO runs. The gas surface density in the spiral arms increases at a lower rate for the RAMSES simulations compared to the other codes. There is also a greater degree of substructure in the SPHNG and GIZMO runs and secondary spiral arms are more pronounced. By resolving the Jeans length with a greater number of grid cells, we achieve more similar results to the Lagrangian codes used in this study. Other alterations to the refinement scheme (adding extra levels of refinement and refining based on local density gradients) are less successful in reducing the disparity between RAMSES and SPHNG/GIZMO. Although more similar, SPHNG displays different density distributions and vertical mass profiles to all modes of GIZMO (including the smoothed particle hydrodynamics version). This suggests differences also arise which are not intrinsic to the particular method but rather due to its implementation. The discrepancies between codes (in particular, the densities reached in the spiral arms) could potentially result in differences in the locations and time-scales for gravitational collapse, and therefore impact star formation activity in more complex galaxy disc simulations.

  18. Testing hydrodynamics schemes in galaxy disc simulations

    NASA Astrophysics Data System (ADS)

    Few, C. G.; Dobbs, C.; Pettitt, A.; Konstandin, L.

    2016-08-01

    We examine how three fundamentally different numerical hydrodynamics codes follow the evolution of an isothermal galactic disc with an external spiral potential. We compare an adaptive mesh refinement code (RAMSES), a smoothed particle hydrodynamics code (sphNG), and a volume-discretised meshless code (GIZMO). Using standard refinement criteria, we find that RAMSES produces a disc that is less vertically concentrated and does not reach such high densities as the sphNG or GIZMO runs. The gas surface density in the spiral arms increases at a lower rate for the RAMSES simulations compared to the other codes. There is also a greater degree of substructure in the sphNG and GIZMO runs and secondary spiral arms are more pronounced. By resolving the Jeans' length with a greater number of grid cells we achieve more similar results to the Lagrangian codes used in this study. Other alterations to the refinement scheme (adding extra levels of refinement and refining based on local density gradients) are less successful in reducing the disparity between RAMSES and sphNG/GIZMO. Although more similar, sphNG displays different density distributions and vertical mass profiles to all modes of GIZMO (including the smoothed particle hydrodynamics version). This suggests differences also arise which are not intrinsic to the particular method but rather due to its implementation. The discrepancies between codes (in particular, the densities reached in the spiral arms) could potentially result in differences in the locations and timescales for gravitational collapse, and therefore impact star formation activity in more complex galaxy disc simulations.

  19. Drug susceptibility testing of nontuberculous mycobacteria.

    PubMed

    van Ingen, Jakko; Kuijper, Ed J

    2014-01-01

    Diseases caused by nontuberculous mycobacteria are emerging in many settings. With an increased number of patients needing treatment, the role of drug susceptibility testing is again in the spotlight. This articles covers the history and methodology of drug susceptibility tests for nontuberculous mycobacteria, but focuses on the correlations between in vitro drug susceptibility, pharmacokinetics and in vivo outcomes of treatment. Among slow-growing nontuberculous mycobacteria, clear correlations have been established for macrolides and amikacin (Mycobacterium avium complex) and for rifampicin (Mycobacterium kansasii). Among rapid-growing mycobacteria, correlations have been established in extrapulmonary disease for aminoglycosides, cefoxitin and co-trimoxazole. In pulmonary disease, correlations are less clear and outcomes of treatment are generally poor, especially for Mycobacterium abscessus. The clinical significance of inducible resistance to macrolides among rapid growers is an important topic. The true role of drug susceptibility testing for nontuberculous mycobacteria still needs to be addressed, preferably within clinical trials. PMID:25340838

  20. Many factors complicate EM susceptibility tests

    NASA Astrophysics Data System (ADS)

    Richards, R. J.

    1982-09-01

    Procedures and apparatus currently employed for assaying the EM susceptibility of communication, navigation, and EW equipment are described. Susceptibility is examined in either conducted susceptibility tests, where signals are introduced into the input port of the device under test, or in radiated modes, where the entire device is exposed to an EM field to test for component and system failure. Noting that military standards require up to 10 times the EM resistance as commercial standards, the use of shielded enclosures in both commercial and military testing facilities is explored. RF-tight enclosures are filled with a homogeneous EM field produced by, optimally, broadband generators which emit signals which are amplified to desired levels. Sweep functions permit testing under broadband conditions. Attention is given to radiator selection and antenna choice to produce satisfactory test conditions at all frequencies.

  1. Antianaerobic Antimicrobials: Spectrum and Susceptibility Testing

    PubMed Central

    Wexler, Hannah M.; Goldstein, Ellie J. C.

    2013-01-01

    SUMMARY Susceptibility testing of anaerobic bacteria recovered from selected cases can influence the choice of antimicrobial therapy. The Clinical and Laboratory Standards Institute (CLSI) has standardized many laboratory procedures, including anaerobic susceptibility testing (AST), and has published documents for AST. The standardization of testing methods by the CLSI allows comparisons of resistance trends among various laboratories. Susceptibility testing should be performed on organisms recovered from sterile body sites, those that are isolated in pure culture, or those that are clinically important and have variable or unique susceptibility patterns. Organisms that should be considered for individual isolate testing include highly virulent pathogens for which susceptibility cannot be predicted, such as Bacteroides, Prevotella, Fusobacterium, and Clostridium spp.; Bilophila wadsworthia; and Sutterella wadsworthensis. This review describes the current methods for AST in research and reference laboratories. These methods include the use of agar dilution, broth microdilution, Etest, and the spiral gradient endpoint system. The antimicrobials potentially effective against anaerobic bacteria include beta-lactams, combinations of beta-lactams and beta-lactamase inhibitors, metronidazole, chloramphenicol, clindamycin, macrolides, tetracyclines, and fluoroquinolones. The spectrum of efficacy, antimicrobial resistance mechanisms, and resistance patterns against these agents are described. PMID:23824372

  2. Antianaerobic antimicrobials: spectrum and susceptibility testing.

    PubMed

    Brook, Itzhak; Wexler, Hannah M; Goldstein, Ellie J C

    2013-07-01

    Susceptibility testing of anaerobic bacteria recovered from selected cases can influence the choice of antimicrobial therapy. The Clinical and Laboratory Standards Institute (CLSI) has standardized many laboratory procedures, including anaerobic susceptibility testing (AST), and has published documents for AST. The standardization of testing methods by the CLSI allows comparisons of resistance trends among various laboratories. Susceptibility testing should be performed on organisms recovered from sterile body sites, those that are isolated in pure culture, or those that are clinically important and have variable or unique susceptibility patterns. Organisms that should be considered for individual isolate testing include highly virulent pathogens for which susceptibility cannot be predicted, such as Bacteroides, Prevotella, Fusobacterium, and Clostridium spp.; Bilophila wadsworthia; and Sutterella wadsworthensis. This review describes the current methods for AST in research and reference laboratories. These methods include the use of agar dilution, broth microdilution, Etest, and the spiral gradient endpoint system. The antimicrobials potentially effective against anaerobic bacteria include beta-lactams, combinations of beta-lactams and beta-lactamase inhibitors, metronidazole, chloramphenicol, clindamycin, macrolides, tetracyclines, and fluoroquinolones. The spectrum of efficacy, antimicrobial resistance mechanisms, and resistance patterns against these agents are described. PMID:23824372

  3. Antimicrobial susceptibility testing for Mycobacterium sp.

    PubMed

    Blanco-Ruano, Delia; Roberts, David M; Gonzalez-Del-Rio, Ruben; Álvarez, Daniel; Rebollo, Ma José; Pérez-Herrán, Esther; Mendoza, Alfonso

    2015-01-01

    The concept of antimicrobial susceptibility testing is an essential part of clinical microbiology. Antimicrobial testing has played a central role in the identification of new antibiotics and defining their clinical uses. Here we describe different approaches to determine the activity of compounds in medium- or high-throughput format. PMID:25779321

  4. Susceptibility testing of Entamoeba histolytica

    SciTech Connect

    Cedeno, J.R.; Krogstad, D.J.

    1983-12-01

    The growth of Entamoeba histolytica in microtiter plates in vitro in a variety of environments with reduced oxygen tensions is reported. With 3% O/sub 2/, 3% CO/sub 2/, and 94% N/sub 2/, the parasite growth in microtiter plates was identical to that in screw-capped culture tubes, as measured by (/sup 3/H)thymidine incorporation and by quantitative parasite counts. There were no significant differences between the drug concentrations necessary to inhibit parasite growth by 50% based on (/sup 3/H)thymidine incorporation vs those defined by quantitative parasite counts for the 15 antimicrobial agents tested (including seven drugs used for the treatment of amebiasis). This technique provides a reproducible method to quantitate the activity of potential antiamebic agents in vitro. The isotopic method should be of particular value in defining the metabolism of the parasite and effects of antimicrobial agents on it, whereas the morphologic method may be more valuable for workers with limited resources available to them.

  5. [Antimicrobial susceptibility testing of anaerobic bacteria].

    PubMed

    García-Sánchez, José E; García-Sánchez, Enrique; García-García, María Inmaculada

    2014-02-01

    The anaerobic bacteria resistance to antibiotics is increasing, and even has appeared against the most active of those, like metronidazol and carbapenems. This fact forces to make and periodical sensibility tests -at least in the most aggressive and virulent species, in cases that they are isolated from life locations and in the absence of therapeutic response- to check the local sensibility and to establish suitable empiric therapies, all based on multicentric studies carried out in order to this or well to check the activity of new antibiotics. For the laboratory routine, the easiest sensibility method is the E-test/MIC evaluator. Another alternative is microdilution, that's only normalized for Bacteroides. There are preliminary facts that allow the use of disc diffusion method in some species of Bacteroides and Clostridium. For the temporal and multicentric studies, the procedure is dilution in agar plate, the reference method. PMID:24630580

  6. Hydrogen gas embrittlement and the disc pressure test

    NASA Technical Reports Server (NTRS)

    Bachelet, E. J.; Troiano, A. R.

    1973-01-01

    A disc pressure test has been used to study the influenced of a hydrogen gas environment on the mechanical properties of three high strength superalloys, Inconel 718, L-605 and A-286, in static and dynamic conditions. The influence of the hydrogen pressure, loading rate, temperature, mechanical and thermal fatigue has investigated. The permeation characteristics of Inconel 718 have been determined in collaboration with the French AEC. The results complemented by a fractographic study are consistent either with a stress-sorption or with an internal embrittlement type of mechanism.

  7. Disk Agar Diffusion Susceptibility Testing of Yeasts

    PubMed Central

    Saubolle, Michael A.; Hoeprich, Paul D.

    1978-01-01

    A disk agar diffusion method was developed for testing the susceptibility of rapidly growing yeasts in vitro. A totally defined, completely synthetic agar culture medium (synthetic amino acid medium, fungal) and clinical isolates of Candida spp. and Torulopsis glabrata were used. Turbidimetric adjustment of cell suspensions resulted in standard, reproducible inocula, which gave sharp, clear zones of inhibition when applied by an agar overlay method. Optimal disk loads were determined for amphotericin B, amphotericin B methyl ester, 5-fluorocytosine, clotrimazole, and miconazole. Disk potencies were stable over a 2-month period when stored in a vacuum desiccator at −30°C. Using an error ratebounded classification, the zones of inhibition were correlated with both broth dilution and agar dilution minimum inhibitory concentrations (MICs). With amphotericin B and amphotericin B methyl ester, all isolates were susceptible, yielding zone diameters which clustered within 5 mm. Overall correlations between zone diameters and broth dilution MICs with 5-fluorocytosine, miconazole, and clotrimazole were 97, 96, and 82% (excluding T. glabrata), respectively; correlations of zone diameters with agar dilution MICs were 96, 92, and 88%, respectively. Disk diffusion susceptibility testing of yeasts appears to be generally applicable. However, when results are equivocal, quantitative test methods should be used. PMID:568910

  8. [An update on antifungal susceptibility testing].

    PubMed

    Tapia P, Cecilia V

    2009-04-01

    Due to increasing of invasive fungal infections and emergeney of antifungal drugs resistant fungi, standardized methods of antifungal susceptibility testing (AST) have been developed. The Clinical Laboratory Standards Instutute (CLSI) and the European for Committee Antimicrobial Susceptibility Testing (EUCAST) have guidelines for susceptibility of yeasts by broth microdilution (M27-A2 and E. Dis. 7.1 documents, respectively). Both are equivalent, although they present methodological and interpretative breakpoints differences. In addition, the CLSI have the M38-A (for filamentous fungi) and M44-A (disk diffusion) documents, whereas EUCAST is developing a document for Aspergillus spp. Furthermore, commercial methods are available that display good correlation with the methods of reference such as E-test, Sensititre and Vitek2. The interpretation of the results must be careful because the determination of the minimum inhibitory concentration (CIM) is difficult for fungi, there are host factors involved and not always there is a correlation between MIC and clinical outcome. Due to these methods are laborious and require trained personnel, to ask for AST to a reference laboratory is recommendable. PMID:19621145

  9. In vitro susceptibility testing of Dientamoeba fragilis.

    PubMed

    Nagata, N; Marriott, D; Harkness, J; Ellis, J T; Stark, D

    2012-01-01

    Dientamoeba fragilis is a commonly encountered trichomonad which has been implicated as a cause of gastrointestinal disease in humans. Despite the frequency of reports recording infections with this parasite, little research has been undertaken in terms of antimicrobial susceptibility. The aim of this study was to evaluate the susceptibility of D. fragilis to several commonly used antiparasitic agents: diloxanide furoate, furazolidone, iodoquinol, metronidazole, nitazoxanide, ornidazole, paromomycin, secnidazole, ronidazole, tetracycline, and tinidazole. Antibiotic susceptibility testing was performed on four clinical strains of D. fragilis, designated A, E, M, and V, respectively. Molecular testing followed, and all strains were determined to be genotype 1. The activities of antiprotozoal compounds at concentrations ranging from 2 μg/ml to 500 μg/ml were determined via cell counts of D. fragilis trophozoites grown in dixenic culture. Minimum lethal concentrations (MLCs) were as follows: ornidazole, 8 to 16 μg/ml; ronidazole, 8 to 16 μg/ml; tinidazole, 31 μg/ml; metronidazole, 31 μg/ml; secnidazole, 31 to 63 μg/ml; nitazoxanide, 63 μg/ml; tetracycline, 250 μg/ml; furazolidone, 250 to 500 μg/ml; iodoquinol, 500 μg/ml; paromomycin, 500 μg/ml; and diloxanide furoate, >500 μg/ml. This is the first study to report the profiles of susceptibility to a wide range of commonly used treatments for clinical isolates of D. fragilis. Our study indicated 5-nitroimidazole derivatives to be the most active compounds in vitro against D. fragilis. PMID:22024820

  10. In Vitro Susceptibility Testing of Dientamoeba fragilis

    PubMed Central

    Nagata, N.; Marriott, D.; Harkness, J.; Ellis, J. T.

    2012-01-01

    Dientamoeba fragilis is a commonly encountered trichomonad which has been implicated as a cause of gastrointestinal disease in humans. Despite the frequency of reports recording infections with this parasite, little research has been undertaken in terms of antimicrobial susceptibility. The aim of this study was to evaluate the susceptibility of D. fragilis to several commonly used antiparasitic agents: diloxanide furoate, furazolidone, iodoquinol, metronidazole, nitazoxanide, ornidazole, paromomycin, secnidazole, ronidazole, tetracycline, and tinidazole. Antibiotic susceptibility testing was performed on four clinical strains of D. fragilis, designated A, E, M, and V, respectively. Molecular testing followed, and all strains were determined to be genotype 1. The activities of antiprotozoal compounds at concentrations ranging from 2 μg/ml to 500 μg/ml were determined via cell counts of D. fragilis trophozoites grown in dixenic culture. Minimum lethal concentrations (MLCs) were as follows: ornidazole, 8 to 16 μg/ml; ronidazole, 8 to 16 μg/ml; tinidazole, 31 μg/ml; metronidazole, 31 μg/ml; secnidazole, 31 to 63 μg/ml; nitazoxanide, 63 μg/ml; tetracycline, 250 μg/ml; furazolidone, 250 to 500 μg/ml; iodoquinol, 500 μg/ml; paromomycin, 500 μg/ml; and diloxanide furoate, >500 μg/ml. This is the first study to report the profiles of susceptibility to a wide range of commonly used treatments for clinical isolates of D. fragilis. Our study indicated 5-nitroimidazole derivatives to be the most active compounds in vitro against D. fragilis. PMID:22024820

  11. Tests and developments of the PANDA Endcap Disc DIRC

    NASA Astrophysics Data System (ADS)

    Etzelmüller, E.; Belias, A.; Dzhygadlo, R.; Gerhardt, A.; Götzen, K.; Kalicy, G.; Krebs, M.; Lehmann, D.; Nerling, F.; Patsyuk, M.; Peters, K.; Schepers, G.; Schmitt, L.; Schwarz, C.; Schwiening, J.; Traxler, M.; Zühlsdorf, M.; Britting, A.; Eyrich, W.; Lehmann, A.; Pfaffinger, M.; Uhlig, F.; Düren, M.; Föhl, K.; Hayrapetyan, A.; Kröck, B.; Merle, O.; Rieke, J.; Schmidt, M.; Cowie, E.; Keri, T.; Achenbach, P.; Cardinali, M.; Hoek, M.; Lauth, W.; Schlimme, S.; Sfienti, C.; Thiel, M.

    2016-04-01

    The PANDA experiment at the future Facility for Antiproton and Ion Research (FAIR) requires excellent particle identification. Two different DIRC detectors will utilize internally reflected Cherenkov light of charged particles to enable the separation of pions and kaons up to momenta of 4 GeV/c. The Endcap Disc DIRC will be placed in the forward endcap of PANDA's central spectrometer covering polar angles between 5° and 22°. Its final design is based on MCP-PMTs for the photon detection and an optical system made of fused silica. A new prototype has been investigated during a test beam at CERN in May 2015 and first results will be presented. In addition a new synthetic fused silica material by Nikon has been tested and was found to be radiation hard.

  12. Awareness of Cancer Susceptibility Genetic Testing

    PubMed Central

    Mai, Phuong L.; Vadaparampil, Susan Thomas; Breen, Nancy; McNeel, Timothy S.; Wideroff, Louise; Graubard, Barry I.

    2014-01-01

    Background Genetic testing for several cancer susceptibility syndromes is clinically available; however, existing data suggest limited population awareness of such tests. Purpose To examine awareness regarding cancer genetic testing in the U.S. population aged ≥25 years in the 2000, 2005, and 2010 National Health Interview Surveys. Methods The weighted percentages of respondents aware of cancer genetic tests, and percent changes from 2000–2005 and 2005–2010, overall and by demographic, family history, and healthcare factors were calculated. Interactions were used to evaluate the patterns of change in awareness between 2005 and 2010 among subgroups within each factor. To evaluate associations with awareness in 2005 and 2010, percentages were adjusted for covariates using multiple logistic regression. The analysis was performed in 2012. Results Awareness decreased from 44.4% to 41.5% (p<0.001) between 2000 and 2005, and increased to 47.0% (p<0.001) in 2010. Awareness increased between 2005 and 2010 in most subgroups, particularly among individuals in the South (p-interaction=0.03) or with a usual place of care (p-interaction=0.01). In 2005 and 2010, awareness was positively associated with personal or family cancer history and high perceived cancer risk, and inversely associated with racial/ethnic minorities, age 25–39 or ≥60 years, male gender, lower education and income levels, public or no health insurance, and no provider contact in 12 months. Conclusions Despite improvement from 2005 to 2010, ≤50% of the U.S. adult population was aware of cancer genetic testing in 2010. Notably, disparities persist for racial/ethnic minorities and individuals with limited health care access or income. PMID:24745633

  13. Reliability of provocative tests of motion sickness susceptibility

    NASA Technical Reports Server (NTRS)

    Calkins, D. S.; Reschke, M. F.; Kennedy, R. S.; Dunlop, W. P.

    1987-01-01

    Test-retest reliability values were derived from motion sickness susceptibility scores obtained from two successive exposures to each of three tests: (1) Coriolis sickness sensitivity test; (2) staircase velocity movement test; and (3) parabolic flight static chair test. The reliability of the three tests ranged from 0.70 to 0.88. Normalizing values from predictors with skewed distributions improved the reliability.

  14. Comparison of an Automated System with Conventional Identification and Antimicrobial Susceptibility Testing

    PubMed Central

    Duggal, Shalini; Gaind, Rajni; Tandon, Neha; Deb, Manorama; Chugh, Tulsi Das

    2012-01-01

    The present study was designed to compare a fully automated identification/antibiotic susceptibility testing (AST) system BD Phoenix (BD) for its efficacy in rapid and accurate identification and AST with conventional manual methods and to determine if the errors reported in AST, such as the (very major errors) VME (false susceptibility), (major errors) ME (false resistance), and (minor errors) MiE (intermediate category interpretation) were within the range certified by FDA. Identification and antimicrobial susceptibility test results of eighty-five clinical isolates including both gram-positive and negative were compared on Phoenix considering the results obtained from conventional manual methods of identification and disc diffusion testing of antibiotics as standards for comparison. Phoenix performed favorably well. There was 100% concordance in identification for gram-negative isolates and 94.83% for gram-positive isolates. In seven cases, Phoenix proved better than conventional identification. For antibiotic results, categorical agreement was 98.02% for gram-positive and 95.7% for gram-negative isolates. VME was 0.33%, ME 0.66%, MiE 0.99% for gram-positive isolates and 1.23% VME, 1.23% ME, and 1.85% MiE for gram-negative isolates. Therefore, this automated system can be used as a tool to facilitate early identification and susceptibility pattern of aerobic bacteria in routine microbiology laboratories. PMID:23762748

  15. [Should metal alloy discs be used for patch testing in suspected metal implant intolerance reaction?].

    PubMed

    Thomas, P; Geier, J; Dickel, H; Diepgen, T; Hillen, U; Kreft, B; Schnuch, A; Szliska, C; Mahler, V

    2015-11-01

    Intolerance reactions to metal implants may be caused by metal allergy. However, prior to implantation, patch testing should not be done in a prophylactic-prophetic approach. Pre-implant patch testing should only be performed to verify or exclude metal allergy in patients with a reported respective history. In the case of implant-in particular arthroplasty-related complications like, for example, pain, effusion, skin changes, reduced range of motion, or loosening, orthopedic-surgical differential diagnostics should be performed first. Allergological workup of suspected metal implant allergy should be done with the DKG baseline series which contains nickel-, cobalt- and chromium-preparations. Various studies assessing the usefulness of metal alloy discs for patch testing proved that this approach does not give reliable information about metal allergy. Positive patch test reactions to the discs cannot be assigned to a specific metal within the disc alloy components. Furthermore, availability of such metal discs might be an invitation to uncritical testing. Accordingly, due to lack of benefit in comparison to patch testing with standardized metal salt preparations, we do not recommend patch testing with metal alloy discs. PMID:26438196

  16. Automatic radiated susceptibility test system for payload equipment

    NASA Astrophysics Data System (ADS)

    Ngo, Hoai T.; Sturman, John C.; Sargent, Noel B.

    1995-09-01

    An automatic radiated susceptibility test system (ARSTS) was developed for NASA Lewis Research Center's Electro-magnetic Interference laboratory. According to MSFC-SPEC 521B, any electrical or electronic equipment that will be transported by the spacelab and space shuttle must be tested for susceptibility to electromagnetic interference. This state-of-the-art automatic test system performs necessary calculations; analyzes, processes, and records a great quantity of measured data; and monitors the equipment being tested in real-time and with minimal user intervention. ARSTS reduces costly test time, increases test accuracy, and provides reliable test results.

  17. Automatic Radiated Susceptibility Test System for Payload Equipment

    NASA Technical Reports Server (NTRS)

    Ngo, Hoai T.; Sturman, John C.; Sargent, Noel B.

    1995-01-01

    An automatic radiated susceptibility test system (ARSTS) was developed for NASA Lewis Research Center's Electro-magnetic Interference laboratory. According to MSFC-SPEC 521B, any electrical or electronic equipment that will be transported by the spacelab and space shuttle must be tested for susceptibility to electromagnetic interference. This state-of-the-art automatic test system performs necessary calculations; analyzes, processes, and records a great quantity of measured data; and monitors the equipment being tested in real-time and with minimal user intervention. ARSTS reduces costly test time, increases test accuracy, and provides reliable test results.

  18. Miniaturized rotating disc rheometer test for rapid screening of drag reducing marine coatings

    NASA Astrophysics Data System (ADS)

    Dennington, Simon; Mekkhunthod, Ponkrit; Rides, Martin; Gibbs, David; Salta, Maria; Stoodley, Victoria; Wharton, Julian; Stoodley, Paul

    2015-09-01

    Frictional drag from the submerged hull surface of a ship is a major component of the resistance experienced when moving through water. Techniques for measuring frictional drag on test surfaces include towing tanks, flow tunnels and rotating discs. These large-scale methods present practical difficulties that hinder their widespread adoption and they are not conducive to rapid throughput. In this study a miniaturized benchtop rotating disc method is described that uses test discs 25 mm in diameter. A highly sensitive analytical rheometer is used to measure the torque acting on the discs rotating in water. Frictional resistance changes are estimated by comparing momentum coefficients. Model rough surfaces were prepared by attaching different grades of sandpaper to the disc surface. Discs with experimental antifouling coatings applied were exposed in the marine environment for the accumulation of microbial fouling, and the rotor was capable of detecting the increased drag due to biofilm formation. The drag due to biofilm was related to an equivalent sand roughness.

  19. Use of a proposed antimicrobial susceptibility testing method for Haemophilus parasuis.

    PubMed

    Dayao, Denise Ann E; Kienzle, Marco; Gibson, Justine S; Blackall, Patrick J; Turni, Conny

    2014-08-27

    The aim of this study was to examine the antimicrobial susceptibility of 97 Haemophilus parasuis cultured from Australian pigs. As there is no existing standard antimicrobial susceptibility technique available for H. parasuis, methods utilising the supplemented media, BA/SN for disc diffusion and test medium broth (TMB) for a microdilution technique, were initially evaluated with the reference strains recommended by the Clinical and Laboratory Standards Institute. The results of the media evaluation suggested that BA/SN and TMB can be used as suitable media for susceptibility testing of H. parasuis. The proposed microdilution technique was then used with 97 H. parasuis isolates and nine antimicrobial agents. The study found that Australian isolates showed elevated minimum inhibitory concentrations (MICs) for ampicillin (1%), penicillin (2%), erythromycin (7%), tulathromycin (9%), tilmicosin (22%), tetracycline (31%) and trimethoprim-sulfamethoxazole (40%). This study has described potential antimicrobial susceptibility methods for H. parasuis and has detected a low percentage of Australian H. parasuis isolates with elevated antimicrobial MICs. PMID:24984947

  20. Estimation of friction under forging conditions by means of the ring-on-disc test

    SciTech Connect

    Buchner, Bernhard; Umgeher, Andreas; Buchmayr, Bruno

    2007-04-07

    In order to understand the tribological processes and interactions in the tool-workpiece-interface systematically, basic experiments that allow an independent variation of influencing parameters are necessary. The ring-on-disc test is a popular model experiment that is often used in tribological analyses at low normal contact pressures.The scope of the paper is an analysis of the applicability of the ring-on-disc test for high normal pressures as used in forging processes, using aluminium AA6082 as workpiece material. It turned out, that this test is a convenient method to measure friction under forging conditions.

  1. [Internal quality control for antimicrobial susceptibility test: an experience feedback].

    PubMed

    Vedy, Serge

    2012-01-01

    Internal quality control (IQC) position in antimicrobial susceptibility testing must be evaluated attentively before using. Ours S. aureus ATCC 25923 use during year 2011 has given precious information that can be useful for other laboratories. First, IQC should never be used before checking that all the realisation process steps are controlled. It will then appear that reagents are the most susceptible to give false results. That's what happens in 2.74% of antimicrobial susceptibility test. IQC is then useful to limit their clinical's impact. However, IQC use also shows that quality improvement will be difficult without industrial producer's involvement. PMID:22826843

  2. Are Creativity Tests Susceptible to Coaching?

    ERIC Educational Resources Information Center

    Fairweather, Elizabeth C.; Cramond, Bonnie; Landis, Rebecca Nordin

    2015-01-01

    Critics of creativity tests, some of which are used widely in determining student eligibility for special educational opportunities such as gifted programs, claim that scores on these tests can be improved with specific training on the scoring components of the tests. However, we could find no empirical evidence supporting this claim. So, we…

  3. Evaluation of five susceptibility test methods for detection of tobramycin resistance in a cluster of epidemiologically related Acinetobacter baumannii isolates.

    PubMed

    Moodley, V Mischka; Oliver, Stephen P; Shankland, Iva; Elisha, B Gay

    2013-08-01

    Acinetobacter baumannii is a major nosocomial pathogen causing infections in critically ill patients. This organism has acquired the propensity to rapidly develop resistance to most antibiotics. At several hospitals within Cape Town, South Africa, tobramycin and colistin are frequently the only therapeutic options. Vitek2 automated susceptibility testing (AST) is used in the clinical laboratory to determine selected susceptibility profiles. The suspicion of a possible AST-related technical error when testing for susceptibility to tobramycin in A. baumannii precipitated this study. Thirty-nine A. baumannii strains isolated from clinical specimens (June to December 2006) were included in this prospective study. Tobramycin susceptibility testing results obtained by AST, disc diffusion, the epsilometer test (Etest), and agar dilution were compared to those for broth microdilution (BMD), the reference method. The tobramycin susceptibility results revealed errors in 25/39 (64%) isolates (10 very major and 15 minor errors) when AST was compared to BMD, 12/39 (31%) (2 very major and 10 minor errors) when Etest was compared to BMD, 16/39 (41%) (3 very major and 13 minor errors) when disc diffusion was compared to BMD, and 21/39 (54%) (10 very major and 11 minor errors) when agar dilution was compared to BMD. Using PCR, we detected aac(3)-IIa, which is associated with tobramycin resistance, in 21/25 of the discrepant isolates. Molecular typing (using pulsed-field gel electrophoresis and repetitive sequence-based PCR [rep-PCR]) showed that these isolates were genetically related. Clinical laboratories that routinely use the Vitek2 system should consider an alternative testing method for determining susceptibility to tobramycin. PMID:23698528

  4. Antifungal Susceptibility Testing of Ascomycetous Yeasts Isolated from Animals.

    PubMed

    Álvarez-Pérez, Sergio; García, Marta E; Peláez, Teresa; Martínez-Nevado, Eva; Blanco, José L

    2016-08-01

    Recent studies suggest that antifungal resistance in yeast isolates of veterinary origin may be an underdiagnosed threat. We tested a collection of 92 ascomycetous yeast isolates that were obtained in Spain from birds, mammals and insects for antifungal susceptibility. MICs to amphotericin B and azoles were low, and no resistant isolates were detected. Despite these results, and given the potential role of animals as reservoirs of resistant strains, continuous monitoring of antifungal susceptibility in the veterinary setting is recommended. PMID:27216048

  5. Susceptibility testing of anaerobic bacteria: myth, magic, or method?

    PubMed Central

    Wexler, H M

    1991-01-01

    The demand for susceptibility testing of anaerobes has increased, yet consensus as to procedure and interpretation in this area has not been achieved. While routine testing of anaerobic isolates is not needed, certain isolates in specific clinical settings should be tested. Also, laboratories may monitor their local antibiograms by doing periodic surveillance batch testing. The National Committee for Clinical Laboratory Standards has published a protocol of methods approved for susceptibility testing of anaerobic bacteria. Both agar and broth microdilution are included; however, the broth disk elution method is no longer approved by the National Committee for Clinical Laboratory Standards because of method-related interpretive errors. A number of newer methods are undergoing evaluation and seem promising. Clinicians and microbiologists reviewing susceptibility reports should be aware of sources of variability in the test results. Variables in susceptibility testing of anaerobes include the media and methods used, organisms chosen for testing, breakpoints chosen for interpretation, antibiotic, and determination of endpoint. Clustering of MICs around the breakpoint may lead to significant variability in test results. Adherence of testing laboratories to approved methods and careful descriptions of the method and the breakpoints used for interpretation would facilitate interlaboratory comparisons and allow problems of emerging resistance to be noted. A variety of resistance mechanisms occurs in anaerobic bacteria, including the production of beta-lactamase and other drug-inactivating enzymes, alteration of target proteins, and inability of the drug to penetrate the bacterial wall. Antimicrobial resistance patterns in the United States and abroad are described. PMID:1747863

  6. Improved method for azole antifungal susceptibility testing.

    PubMed Central

    Gordon, M A; Lapa, E W; Passero, P G

    1988-01-01

    A reproducible method is described for the determination of the MICs of ketoconazole, miconazole, fluconazole, and itraconazole with sharp endpoints when employed with either yeasts or molds. A semisolid medium is used with controlled pH and standardized inoculum. The time of reading results is a critical factor in the conduct of this test. The medium is simple to prepare and has a relatively long refrigerator shelf life in a user-ready state, requiring only the addition of a freshly prepared inoculum after restoration to room temperature. Images PMID:2846651

  7. Population genetic testing for cancer susceptibility: founder mutations to genomes.

    PubMed

    Foulkes, William D; Knoppers, Bartha Maria; Turnbull, Clare

    2016-01-01

    The current standard model for identifying carriers of high-risk mutations in cancer-susceptibility genes (CSGs) generally involves a process that is not amenable to population-based testing: access to genetic tests is typically regulated by health-care providers on the basis of a labour-intensive assessment of an individual's personal and family history of cancer, with face-to-face genetic counselling performed before mutation testing. Several studies have shown that application of these selection criteria results in a substantial proportion of mutation carriers being missed. Population-based genetic testing has been proposed as an alternative approach to determining cancer susceptibility, and aims for a more-comprehensive detection of mutation carriers. Herein, we review the existing data on population-based genetic testing, and consider some of the barriers, pitfalls, and challenges related to the possible expansion of this approach. We consider mechanisms by which population-based genetic testing for cancer susceptibility could be delivered, and suggest how such genetic testing might be integrated into existing and emerging health-care structures. The existing models of genetic testing (including issues relating to informed consent) will very likely require considerable alteration if the potential benefits of population-based genetic testing are to be fully realized. PMID:26483301

  8. 21 CFR 866.1640 - Antimicrobial susceptibility test powder.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... 21 Food and Drugs 8 2010-04-01 2010-04-01 false Antimicrobial susceptibility test powder. 866.1640 Section 866.1640 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) MEDICAL DEVICES IMMUNOLOGY AND MICROBIOLOGY DEVICES Diagnostic Devices § 866.1640...

  9. 21 CFR 866.1640 - Antimicrobial susceptibility test powder.

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ... 21 Food and Drugs 8 2013-04-01 2013-04-01 false Antimicrobial susceptibility test powder. 866.1640 Section 866.1640 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) MEDICAL DEVICES IMMUNOLOGY AND MICROBIOLOGY DEVICES Diagnostic Devices § 866.1640...

  10. 21 CFR 866.1640 - Antimicrobial susceptibility test powder.

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ... 21 Food and Drugs 8 2014-04-01 2014-04-01 false Antimicrobial susceptibility test powder. 866.1640 Section 866.1640 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) MEDICAL DEVICES IMMUNOLOGY AND MICROBIOLOGY DEVICES Diagnostic Devices § 866.1640...

  11. 21 CFR 866.1640 - Antimicrobial susceptibility test powder.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... 21 Food and Drugs 8 2011-04-01 2011-04-01 false Antimicrobial susceptibility test powder. 866.1640 Section 866.1640 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) MEDICAL DEVICES IMMUNOLOGY AND MICROBIOLOGY DEVICES Diagnostic Devices § 866.1640...

  12. 21 CFR 866.1640 - Antimicrobial susceptibility test powder.

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ... 21 Food and Drugs 8 2012-04-01 2012-04-01 false Antimicrobial susceptibility test powder. 866.1640 Section 866.1640 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) MEDICAL DEVICES IMMUNOLOGY AND MICROBIOLOGY DEVICES Diagnostic Devices § 866.1640...

  13. Mechanical testing and modelling of carbon-carbon composites for aircraft disc brakes

    NASA Astrophysics Data System (ADS)

    Bradley, Luke R.

    The objective of this study is to improve the understanding of the stress distributions and failure mechanisms experienced by carbon-carbon composite aircraft brake discs using finite element (FE) analyses. The project has been carried out in association with Dunlop Aerospace as an EPSRC CASE studentship. It therefore focuses on the carbon-carbon composite brake disc material produced by Dunlop Aerospace, although it is envisaged that the approach will have broader applications for modelling and mechanical testing of carbon-carbon composites in general. The disc brake material is a laminated carbon-carbon composite comprised of poly(acrylonitrile) (PAN) derived carbon fibres in a chemical vapour infiltration (CVI) deposited matrix, in which the reinforcement is present in both continuous fibre and chopped fibre forms. To pave the way for the finite element analysis, a comprehensive study of the mechanical properties of the carbon-carbon composite material was carried out. This focused largely, but not entirely, on model composite materials formulated using structural elements of the disc brake material. The strengths and moduli of these materials were measured in tension, compression and shear in several orientations. It was found that the stress-strain behaviour of the materials were linear in directions where there was some continuous fibre reinforcement, but non-linear when this was not the case. In all orientations, some degree of non-linearity was observed in the shear stress-strain response of the materials. However, this non-linearity was generally not large enough to pose a problem for the estimation of elastic moduli. Evidence was found for negative Poisson's ratio behaviour in some orientations of the material in tension. Additionally, the through-thickness properties of the composite, including interlaminar shear strength, were shown to be positively related to bulk density. The in-plane properties were mostly unrelated to bulk density over the range of

  14. In vitro susceptibility testing of Paracoccidioides brasiliensis to sulfonamides.

    PubMed Central

    Restrepo, A; Arango, M D

    1980-01-01

    A total of 60 clinical isolates of Paracoccidioides brasiliensis were tested for susceptibility to sulfadiazine and sulfadimethoxyne by the agar dilution technique. A modification of the Mueller-Hinton medium was devised which gave good growth of the yeast form. The minimum inhibitory concentrations for only 51.6% of the isolates were in the range of the recommended blood serum concentration (50 micrograms/ml). For 6 to 8% of the isolates, the minimum inhibitory concentrations were above 200 micrograms of both sulfadiazine and sulfadimethoxyne per ml. A significant decreases in susceptibility was demonstrated for one isolate obtained from a patient relapsing during sulfonamide therapy. Images PMID:7416744

  15. A test of truncation in the accretion discs of X-ray Binaries.

    NASA Astrophysics Data System (ADS)

    Eckersall, A.

    2016-06-01

    The truncated-disc model is generally used to help explain the change between the soft and hard states in X-ray Binaries, where the standard accretion disc is truncated in the inner regions and replaced by a radiatively inefficient accretion flow. There is still disagreement though in the extent of this truncation, particularly in at what point truncation begins. Here we analyze XMM EPIC-pn spectra in both the soft and hard states for a number of galactic XRBs, along with RGS data and the latest absorption and emission models to get an independent fit for the ISM column densities for each source. Specifically, we assume the 'canonical' model where the luminous accretion disc extends down to the innermost stable orbit at 6r_g, and construct a spectral model accounting for thermal, reflection and Compton processes ensuring consistent geometrical properties of the models. Rather than attempting to infer the inner disc location from spectral fitting and/or reflection models, we instead attempt a direct test of whether a consistent model will fit assuming no truncation. We discuss the implications for emission models of XRBs.

  16. [Susceptibilities of clinical bacterial isolates to antimicrobial agents. A study mainly focused on imipenem. Research Group for Testing Imipenem Susceptibility on Clinical Isolates].

    PubMed

    Igari, J

    1990-10-01

    We investigated susceptibilities of clinical bacterial isolates to imipenem (IPM) and other antimicrobial agents at 459 hospital laboratories throughout Japan from September to December of 1988. In this study, identification and susceptibility testing were performed at each hospital laboratory and the tests were carried out according to the 1-dilution or 3-dilution disc technique in which susceptibilities are classified into 4 grades: , ++, + and -. IPM had significantly high activity against Streptococcus pneumoniae, Streptococcus pyogenes, Streptococcus agalactiae, Neisseria gonorrhoeae, Escherichia coli, Klebsiella pneumoniae, Klebsiella oxytoca, Enterobacter aerogenes, Enterobacter cloacae, Salmonella spp., Citrobacter freundii, Proteus mirabilis, Providencia rettgeri, Acinetobacter calcoaceticus, Moraxella catarrhalis, Alcaligenes spp., Peptococcus spp./Peptostreptococcus spp., Bacteroides fragilis and Bacteroides spp. and should slightly lower activities on coagulase-negative staphylococci (CNS), Enterococcus faecalis, Haemophilus influenzae, Serratia marcescens, Proteus vulgaris, Providencia stuartii and Pseudomonas aeruginosa than on the above mentioned bacteria. In a comparative study on activities of IPM against bacteria from different clinical sources, no remarkable differences were found due to different sources among S. pneumoniae, E. faecalis, H. influenzae, E. coli, K. pneumoniae, E. cloacae, C. freundii, P. mirabilis or A. calcoaceticus, whereas slight differences were found among Staphylococcus aureus, CNS, S. marcescens and P. aeruginosa. PMID:2086814

  17. Digital antimicrobial susceptibility testing using the MilliDrop technology.

    PubMed

    Jiang, L; Boitard, L; Broyer, P; Chareire, A-C; Bourne-Branchu, P; Mahé, P; Tournoud, M; Franceschi, C; Zambardi, G; Baudry, J; Bibette, J

    2016-03-01

    We present the MilliDrop Analyzer (MDA), a droplet-based millifluidic system for digital antimicrobial susceptibility testing (D-AST), which enables us to determine minimum inhibitory concentrations (MICs) precisely and accurately. The MilliDrop technology was validated by using resazurin for fluorescence readout, for comparison with standard methodology, and for conducting reproducibility studies. In this first assessment, the susceptibility of a reference Gram-negative strain Escherichia coli ATCC 25922 to gentamicin, chloramphenicol, and nalidixic acid were tested by the MDA, VITEK®2, and broth microdilution as a reference standard. We measured the susceptibility of clinically relevant Gram-positive strains of Staphylococcus aureus to vancomycin, including vancomycin-intermediate S. aureus (VISA), heterogeneous vancomycin-intermediate S. aureus (hVISA), and vancomycin-susceptible S. aureus (VSSA) strains. The MDA provided results which were much more accurate than those of VITEK®2 and standard broth microdilution. The enhanced accuracy enabled us to reliably discriminate between VSSA and hVISA strains. PMID:26803821

  18. Genetic susceptibility testing for neurodegenerative diseases: Ethical and practice issues

    PubMed Central

    Roberts, J. Scott; Uhlmann, Wendy R.

    2013-01-01

    As the genetics of neurodegenerative disease become better understood, opportunities for genetic susceptibility testing for at-risk individuals will increase. Such testing raises important ethical and practice issues related to test access, informed consent, risk estimation and communication, return of results, and policies to prevent genetic discrimination. The advent of direct-to-consumer genetic susceptibility testing for various neurodegenerative disorders (including Alzheimer’s disease, Parkinson’s disease, and certain prion diseases) means that ethical and practical challenges must be faced not only in traditional research and clinical settings, but also in broader society. This review addresses several topics relevant to the development and implementation of genetic susceptibility tests across research, clinical, and consumer settings; these include appropriate indications for testing, the implications of different methods for disclosing test results, clinical versus personal utility of risk information, psychological and behavioral responses to test results, testing of minors, genetic discrimination, and ethical dilemmas posed by whole-genome sequencing. We also identify future areas of likely growth in the field, including pharmacogenomics and genetic screening for individuals considering or engaged in activities that pose elevated risk of brain injury (e.g., football players, military personnel). APOE gene testing for risk of Alzheimer’s disease is used throughout as an instructive case example, drawing upon the authors’ experience as investigators in a series of multisite randomized clinical trials that have examined the impact of disclosing APOE genotype status to interested individuals (e.g., first-degree relatives, persons with mild cognitive impairment). PMID:23583530

  19. Susceptibility testing with the sensititer breakpoint broth microdilution system.

    PubMed

    Doern, G V; Dascal, A; Keville, M

    1985-05-01

    The antimicrobial susceptibility profiles of a total of 318 aerobic and facultatively anaerobic bacteria (255 gram-negative bacilli and 63 gram-positive cocci) were determined, using a new commercially available breakpoint broth microdilution procedure (Sensititer Breakpoint System (SBS), Gibco Diagnostics, Inc., Madison, WI) that categorizes test results in the form of susceptibility categories: susceptible, intermediate, and resistant. Results obtained with the SBS were compared with those achieved with a standardized disk diffusion procedure. Among a total of 4,414 organism-antimicrobic comparisons, concordance between the results of the SBS and the disk diffusion procedure was observed in 3,888 cases (88.1%). Four hundred twenty-three (9.6%) minor discrepancies, 45 (1.0%) major discrepancies, and 58 (1.3%) very major discrepancies were noted. Arbitration of major and very major discrepancies with a full-range minimum inhibitory concentration (MIC) procedure confirmed the results of the SBS in 53.4% of cases. A single organism-antimicrobial combination, the nonenterococcal streptococci tested against the aminoglycosides, yielded a significant number of very major errors which were arbitrated in favor of the disk diffusion result. These errors were probably due to poor growth of the test organism in the broth medium used for performing the SBS test (i.e., cation-supplemented Mueller-Hinton broth). With this exception, the SBS was found to be at least as accurate as the standardized disk diffusion procedure. PMID:3922668

  20. Bacterial nanoscale cultures for phenotypic multiplexed antibiotic susceptibility testing.

    PubMed

    Weibull, Emilie; Antypas, Haris; Kjäll, Peter; Brauner, Annelie; Andersson-Svahn, Helene; Richter-Dahlfors, Agneta

    2014-09-01

    An optimal antimicrobial drug regimen is the key to successful clinical outcomes of bacterial infections. To direct the choice of antibiotic, access to fast and precise antibiotic susceptibility profiling of the infecting bacteria is critical. We have developed a high-throughput nanowell antibiotic susceptibility testing (AST) device for direct, multiplexed analysis. By processing in real time the optical recordings of nanoscale cultures of reference and clinical uropathogenic Escherichia coli strains with a mathematical algorithm, the time point when growth shifts from lag phase to early logarithmic phase (Tlag) was identified for each of the several hundreds of cultures tested. Based on Tlag, the MIC could be defined within 4 h. Heatmap presentation of data from this high-throughput analysis allowed multiple resistance patterns to be differentiated at a glance. With a possibility to enhance multiplexing capacity, this device serves as a high-throughput diagnostic tool that rapidly aids clinicians in prescribing the optimal antibiotic therapy. PMID:24989602

  1. Susceptibility testing of Propionibacterium acnes comparing agar dilution with E test.

    PubMed Central

    Smith, M A; Alperstein, P; France, K; Vellozzi, E M; Isenberg, H D

    1996-01-01

    Propionibacterium acnes has been identified as a significant agent of nosocomial infections, including endophthalmitis. Data concerning susceptibility of P. acnes to newer beta-lactam antibiotics and fluoroquinolones are limited. Recent reports suggest that quinolones have activity against these organisms sufficient to warrant further study. We undertook a study to select appropriate antimicrobial agents for use in a rabbit model of P. acnes endophthalmitis. We compared the antibiotic susceptibilities of P. acnes by using the National Committee for Clinical Laboratory Standards method of agar dilution with the E test. Thirteen clinical isolates obtained from eye specimens and three American Type Culture Collection control strains were tested against 14 antibiotics. All the clinical isolates were susceptible by both methods to piperacillin, piperacillin-tazobactam, ampicillin-sulbactam, ticarcillin-clavulanate, cefotaxime, cefotetan, ceftriaxone, cefoxitin, and imipenem in addition to clindamycin but were resistant to metronidazole. The clinical P. acnes isolates also displayed high-level susceptibility to ciprofloxacin, sparfloxacin, and ofloxacin. Almost all the P. acnes strains demonstrated E-test MICs within 2 dilutions of the MICs observed by the agar dilution method. Those few strains for which discrepancies were noted exhibited E-test susceptibilities three- to fivefold dilutions lower than the agar dilution method susceptibilities but only with ampicillin-sulbactam, ticarcillin-clavulanate, and/or clindamycin. On the basis of our study, all of clinical eye isolates were susceptible to these newer antimicrobial agents and the two methods demonstrated similar susceptibility patterns. PMID:8815076

  2. Resistance mechanisms and drug susceptibility testing of nontuberculous mycobacteria.

    PubMed

    van Ingen, Jakko; Boeree, Martin J; van Soolingen, Dick; Mouton, Johan W

    2012-06-01

    Nontuberculous mycobacteria (NTM) are increasingly recognized as causative agents of opportunistic infections in humans. For most NTM infections the therapy of choice is drug treatment, but treatment regimens differ by species, in particular between slow (e.g. Mycobacterium avium complex, Mycobacterium kansasii) and rapid growers (e.g. Mycobacterium abscessus, Mycobacterium fortuitum). In general, drug treatment is long, costly, and often associated with drug-related toxicities; outcome of drug treatment is poor and is likely related to the high levels of natural antibiotic resistance in NTM. The role of drug susceptibility testing (DST) in the choice of agents for antimicrobial treatment of NTM disease, mainly that by slow growers, remains subject of debate. There are important discrepancies between drug susceptibility measured in vitro and the activity of the drug observed in vivo. In part, these discrepancies derive from laboratory technical issues. There is still no consensus on a standardized method. With the increasing clinical importance of NTM disease, DST of NTM is again in the spotlight. This review provides a comprehensive overview of the mechanisms of drug resistance in NTM, phenotypic methods for testing susceptibility in past and current use for DST of NTM, as well as molecular approaches to assess drug resistance. PMID:22525524

  3. Antifungal Susceptibility Testing: Practical Aspects and Current Challenges

    PubMed Central

    Rex, John H.; Pfaller, Michael A.; Walsh, Thomas J.; Chaturvedi, Vishnu; Espinel-Ingroff, Ana; Ghannoum, Mahmoud A.; Gosey, Linda L.; Odds, Frank C.; Rinaldi, Michael G.; Sheehan, Daniel J.; Warnock, David W.

    2001-01-01

    Development of standardized antifungal susceptibility testing methods has been the focus of intensive research for the last 15 years. Reference methods for yeasts (NCCLS M27-A) and molds (M38-P) are now available. The development of these methods provides researchers not only with standardized methods for testing but also with an understanding of the variables that affect interlaboratory reproducibility. With this knowledge, we have now moved into the phase of (i) demonstrating the clinical value (or lack thereof) of standardized methods, (ii) developing modifications to these reference methods that address specific problems, and (iii) developing reliable commercial test kits. Clinically relevant testing is now available for selected fungi and drugs: Candida spp. against fluconazole, itraconazole, flucytosine, and (perhaps) amphotericin B; Cryptococcus neoformans against (perhaps) fluconazole and amphotericin B; and Aspergillus spp. against (perhaps) itraconazole. Expanding the range of useful testing procedures is the current focus of research in this area. PMID:11585779

  4. Precision of spinal radiographs as a screening test for intervertebral disc calcification in Dachshunds.

    PubMed

    Rosenblatt, Alana J; Hill, Peter B; Davies, Sarah E; Webster, Natalie S; Lappalainen, Anu K; Bottema, Cynthia D K; Caraguel, Charles G B

    2015-11-01

    Among dog breeds, the Dachshund has the highest lifetime incidence of intervertebral disc disease (IVDD). Intervertebral disc (IVD) calcification is an indicator of severe degeneration that predisposes to disc herniation. IVDD is heritable in Dachshunds, and in some countries, breeding candidates are screened to reduce IVDD occurrence by selecting dogs according to their score of radiographically detectable intervertebral disc calcification (RDIDC) and excluding dogs with ≥5 RDIDCs from breeding. This study evaluated the precision of scoring spinal radiographs for IVD calcification and subsequent classification of Dachshund dogs for breeding based on their RDIDC score. Digital radiographs of the spine were obtained in 19 clinically healthy, young adult Dachshunds, and scored for RDIDC independently by five scorers with varying levels of experience, three times each. Within scorer (repeatability) and between scorer (reproducibility) variability was estimated both at the individual IVD level and at the whole dog level for breeding classification purposes. At the IVD level, some degree of scorer effect was supported by the pairwise repeatability (92.3%; 95% CI: 88.8-94.7%) being marginally higher than the reproducibility (89.2%; 95% CI: 85.7-91.8%). Scorer-specific patterns confirmed the presence of scorer subjectivity. Repeatability significantly increased with scorer experience but the reproducibility did not. RDIDC scoring repeatability and reproducibility substantially decreased at the cervicothoracic spine region, likely due to anatomical superimpositions. At the dog level, a breeding classification could be repeated by the same scorer for 83.6% (95% CI: 73.8-90.2%) of the dogs, and was reproduced between two scorers for 80.2% (95% CI: 66.6-89.1%) of the dogs. The repeatability of breeding classification also seemed to increase with scorer experience but not the reproducibility. Overall, RDIDC scoring revealed some degree of inconsistency explained by scorer

  5. Susceptibility testing of Mycobacterium abscessus by isothermal microcalorimetry.

    PubMed

    Boillat-Blanco, Noémie; Furustrand Tafin, Ulrika; Jaton, Katia; Trampuz, Andrej

    2015-10-01

    We evaluated a new method for susceptibility testing of a rapidly growing mycobacterium using real-time measurement of heat (microcalorimetry). MICs of 2 clinical Mycobacterium abscessus isolates were determined by microbroth dilution and E-test. For microcalorimetry, Middlebrook-7H10 agar+10% oleic acid-albumin-dextrose-catalase, containing amikacin, clarithromycin, linezolid, and ciprofloxacin was inoculated with ~10(5)CFU/mL. Heat production was measured at 37°C for 72h. Minimal heat inhibition concentration (MHIC) was defined as the lowest antibiotic concentration inhibiting growth-related heat production. Growth of M. abscessus was detected after a median of 16.5h (range, 8.5-26.9h). Heat detection was proportionally delayed with increasing concentration of antibiotics. MHICs for the tested strains were 16 to >16mg/L for amikacin, >8mg/L for clarithromycin, 4 to >16mg/L for ciprofloxacin, 24 to >32mg/L for linezolid. MHICs were in agreement within two 2-fold dilutions with conventional MICs. Microcalorimetry may accelerate antimicrobial susceptibility testing in mycobacteria and provide additional real-time information on the drug effect. PMID:26210204

  6. Quantifying weld solidification cracking susceptibility using the varestraint test

    SciTech Connect

    Lin, W.; Lippold, J.C.; Nelson, T.W.

    1994-12-31

    Since the introduction of the original Varestraint concept in the 1960`s, the longitudinal- and transverse-type Varestraint tests have become the most widely utilized techniques for quantifying weld solidification cracking susceptibility. Conventionally, cracking susceptibility is assessed by threshold strain to cause cracking and the degree of cracking as quantified by total crack strain to cause cracking and the degree of cracking as quantified by total crack length or maximum crack length. Although material-specific quantifications such as the brittle temperature range (BTR) have been proposed for the transverse-type test, similar quantifications have not been developed for the longitudinal type test. Various alloys including 304, 310, 316L, A-286, AL6XN, 20Cb-3, RA253, and RA333 stainless steels, 625, 690, and 718 nickel-base alloys, 2090, 2219, 5083, and 6061 aluminum alloys were investigated using both longitudinal- and transverse-type Varestraint tests. Tests were performed using a newly developed, computer-controlled Varestraint unit equipped with a 3-axis movable torch, spring-loaded fixture and a servo-hydraulic loading system. It was found that extensive cracking was observed in the fusion zone emanating radially from the solid-liquid inteface toward the fusion boundary in the longitudinal-type test, while weld centerline cracking was prevalent in the transverse-type test. The theoretical basis for the formation of the CSR is that liquation-related cracking only occurs in a certain temperature range known as the BTR. The detailed procedure in the development of the CSR in the fusion zone is described and discussed. This approach allows a weldability data base to be created and the comparison of results from different laboratories using different test techniques.

  7. Post-irradiation examinations and high-temperature tests on undoped large-grain UO2 discs

    NASA Astrophysics Data System (ADS)

    Noirot, J.; Pontillon, Y.; Yagnik, S.; Turnbull, J. A.

    2015-07-01

    Within the Nuclear Fuel Industry Research (NFIR) programme, several fuel variants -in the form of thin circular discs - were irradiated in the Halden Boiling Water Reactor (HBWR) at burn-ups up to ∼100 GWd/tHM. The design of the fuel assembly was similar to that used in other HBWR programmes: the assembly contained several rods with fuel discs sandwiched between Mo discs, which limited temperature differences within each fuel disc. One such variant was made of large-grain UO2 discs (3D grain size = ∼45 μm) which were subjected to three burn-ups: 42, 72 and 96 GWd/tHM. Detailed characterizations of some of these irradiated large-grain UO2 discs were performed in the CEA Cadarache LECA-STAR hot laboratory. The techniques used included electron probe microanalysis (EPMA), scanning electron microscopy (SEM) and secondary ion mass spectrometry (SIMS). Comparisons were then carried out with more standard grain size UO2 discs irradiated under the same conditions. Examination of the high burn-up large-grain UO2 discs revealed the limited formation of a high burn-up structure (HBS) when compared with the standard-grain UO2 discs at similar burn-up. High burn-up discs were submitted to temperature transients up to 1200 °C in the heating test device called Merarg at a relatively low temperature ramp rate (0.2 °C/s). In addition to the total gas release during these tests, the release peaks throughout the temperature ramp were monitored. Tests at 1600 °C were also conducted on the 42 GWd/tHM discs. The fuels were then characterized with the same microanalysis techniques as those used before the tests, to investigate the effects of these tests on the fuel's microstructure and on the fission gas behaviour. This paper outlines the high resistance of this fuel to gas precipitation at high temperature and to HBS formation at high burn-up. It also shows the similarity of the positions, within the grains, where HBS forms at high burn-up and where bubbles appear during the low

  8. Rolling contact fatigue of surface modified 440C using a 'Ge-Polymet' type disc rod test rig

    SciTech Connect

    Thom, R.L.

    1989-03-01

    Through hardened 440 C martensitic stainless steel test specimens were surface modified and tested for changes in rolling contact fatigue using a disc on rod test rig. The surface modifications consisted of nitrogen, boron, titanium, chromium, tantalum, carbon, or molybdenum ion implantation at various ion fluences and energies. Tests were also performed on specimens reactively sputtered with titanium nitride.

  9. Rolling contact fatigue of surface modified 440C using a 'Ge-Polymet' type disc rod test rig

    NASA Technical Reports Server (NTRS)

    Thom, Robert L.

    1989-01-01

    Through hardened 440 C martensitic stainless steel test specimens were surface modified and tested for changes in rolling contact fatigue using a disc on rod test rig. The surface modifications consisted of nitrogen, boron, titanium, chromium, tantalum, carbon, or molybdenum ion implantation at various ion fluences and energies. Tests were also performed on specimens reactively sputtered with titanium nitride.

  10. Localized strain measurements of the intervertebral disc annulus during biaxial tensile testing.

    PubMed

    Karakolis, Thomas; Callaghan, Jack P

    2015-01-01

    Both inter-lamellar and intra-lamellar failures of the annulus have been described as potential modes of disc herniation. Attempts to characterize initial lamellar failure of the annulus have involved tensile testing of small tissue samples. The purpose of this study was to evaluate a method of measuring local surface strains through image analysis of a tensile test conducted on an isolated sample of annular tissue in order to enhance future studies of intervertebral disc failure. An annulus tissue sample was biaxial strained to 10%. High-resolution images captured the tissue surface throughout testing. Three test conditions were evaluated: submerged, non-submerged and marker. Surface strains were calculated for the two non-marker conditions based on motion of virtual tracking points. Tracking algorithm parameters (grid resolution and template size) were varied to determine the effect on estimated strains. Accuracy of point tracking was assessed through a comparison of the non-marker conditions to a condition involving markers placed on tissue surface. Grid resolution had a larger effect on local strain than template size. Average local strain error ranged from 3% to 9.25% and 0.1% to 2.0%, for the non-submerged and submerged conditions, respectively. Local strain estimation has a relatively high potential for error. Submerging the tissue provided superior strain estimates. PMID:25145810

  11. Abrasive Wear Resistance of Tool Steels Evaluated by the Pin-on-Disc Testing

    NASA Astrophysics Data System (ADS)

    Bressan, José Divo; Schopf, Roberto Alexandre

    2011-05-01

    Present work examines tool steels abrasion wear resistance and the abrasion mechanisms which are one main contributor to failure of tooling in metal forming industry. Tooling used in cutting and metal forming processes without lubrication fails due to this type of wear. In the workshop and engineering practice, it is common to relate wear resistance as function of material hardness only. However, there are others parameters which influences wear such as: fracture toughness, type of crystalline structure and the occurrence of hard precipitate in the metallic matrix and also its nature. In the present investigation, the wear mechanisms acting in tool steels were analyzed and, by normalized tests, wear resistance performance of nine different types of tool steels were evaluated by pin-on-disc testing. Conventional tool steels commonly used in tooling such as AISI H13 and AISI A2 were compared in relation to tool steels fabricated by sintering process such as Crucible CPM 3V, CPM 9V and M4 steels. Friction and wear testing were carried out in a pin-on-disc automated equipment which pin was tool steel and the counter-face was a abrasive disc of silicon carbide. Normal load of 5 N, sliding velocity of 0.45 m/s, total sliding distance of 3000 m and room temperature were employed. The wear rate was calculated by the Archard's equation and from the plotted graphs of pin cumulated volume loss versus sliding distance. Specimens were appropriately heat treated by quenching and three tempering cycles. Percentage of alloying elements, metallographic analyses of microstructure and Vickers microhardness of specimens were performed, analyzed and correlated with wear rate. The work is concluded by the presentation of a rank of tool steel wear rate, comparing the different tool steel abrasion wear resistance: the best tool steel wear resistance evaluated was the Crucible CPM 9V steel.

  12. A New Approach for Pyrazinamide Susceptibility Testing in Mycobacterium tuberculosis

    PubMed Central

    Loli, Sebastian; Gilman, Robert H.; Gutierrez, Andrés; Fuentes, Patricia; Cotrina, Milagros; Kirwan, Daniela; Sheen, Patricia

    2012-01-01

    Background: Pyrazinamide (PZA) is an important drug in the treatment of tuberculosis. Microbiological methods of PZA susceptibility testing are controversial and have low reproducibility. After conversion of PZA into pyrazinoic acid (POA) by the bacterial pyrazinamidase enzyme, the drug is expelled from the bacteria by an efflux pump. Objective: To evaluate the rate of POA extrusion from Mycobacterium tuberculosis as a parameter to detect PZA resistance. Methods: The rate of POA extrusion and PZA susceptibility determined by BACTEC 460 were measured for 34 strains in a previous study. PZA resistance was modeled in a logistic regression with the pyrazinoic efflux rate. Result: POA efflux rate predicted PZA resistance with 70.83%–92.85% sensitivity and 100% specificity compared with BACTEC 460. Conclusion: POA efflux rate could be a useful tool for predicting PZA resistance in M. tuberculosis. Further exploration of this approach may lead to the development of new tools for diagnosing PZA resistance, which may be of public health importance. PMID:22372927

  13. DVD technology-based molecular diagnosis platform: quantitative pregnancy test on a disc.

    PubMed

    Li, Xiaochun; Weng, Samuel; Ge, Bixia; Yao, Zhihui; Yu, Hua-Zhong

    2014-05-21

    A diagnosis platform based entirely on DVD technology was developed for on-site quantitation of molecular analytes of interest, e.g., human chorionic gonadotropin (hCG) in urine samples ("quantitative pregnancy test on a disc"). An hCG-specific monoclonal antibody-binding assay prepared on a regular DVD-R was labeled with nanogold-streptavidin conjugates for signal enhancement with a customized silver-staining protocol. An unmodified, conventional computer optical drive was used for assay reading, and free disc-quality analysis software for data processing. The performance (sensitivity and selectivity) of this DVD assay is comparable to that of well-established colorimetric methods (determination of optical darkness ratios) and standard enzyme-linked immunosorbent assays (ELISA). As validated by examining its linear correlation with the ELISA results on the same set of samples, the DVD assay promises to be a low-cost, multiplex, point-of-care (POC) diagnostic tool for physicians and even for individuals at home, producing prompt results. PMID:24695902

  14. Characterization of Renibacterium salmoninarum with reduced susceptibility to macrolide antibiotics by a standardized antibiotic susceptibility test.

    PubMed

    Rhodes, Linda D; Nguyen, Oanh T; Deinhard, Rebecca K; White, Teresa M; Harrell, Lee W; Roberts, Marilyn C

    2008-08-01

    Three cohorts of juvenile and subadult Chinook salmon Oncorhynchus tshawytscha received multiple treatments with macrolide antibiotics for bacterial kidney disease (BKD) during rearing in a captive broodstock program. A total of 77 mortalities among the cohorts were screened for Renibacterium salmoninarum, the etiologic agent of BKD, by agar culture from kidney, and isolates from 7 fish were suitable for growth testing in the presence of macrolide antibiotics. The minimum inhibitory concentration (MIC) of erythromycin and azithromycin was determined by a modification of the standardized broth assay using defined medium. The American Type Culture Collection (ATCC) type strain 33209 exhibited a MIC of 0.008 microg m(-1) to either erythromycin or azithromycin. Isolates from 3 fish displayed MICs identical to the MICs for the ATCC type strain 33209. In contrast, isolates from 4 fish exhibited higher MICs, ranging between 0.125 and 0.250 microg ml(-1) for erythromycin and between 0.016 and 0.031 microg ml(-1) for azithromycin. Sequence analysis of the mutational hotspots for macrolide resistance in the 23S rDNA gene and the open reading frames of ribosomal proteins L4 and L22 found identical sequences among all isolates, indicating that the phenotype was not due to mutations associated with the drug-binding site of 23S rRNA. These results are the first report of R. salmoninarum with reduced susceptibility to macrolide antibiotics isolated from fish receiving multiple antibiotic treatments. PMID:18814542

  15. Laboratory tests to study the influence of rock stress confinement on the performances of TBM discs in tunnels

    NASA Astrophysics Data System (ADS)

    Innaurato, N.; Oggeri, C.; Oreste, P.; Vinai, R.

    2011-06-01

    To clarify some aspects of rock destruction with a disc acting on a high confined tunnel face, a series of tests were carried out to examine fracture mechanisms under an indenter that simulates the tunnel boring machine (TBM) tool action, in the presence of an adjacent groove, when a state of stress (lateral confinement) is imposed on a rock sample. These tests proved the importance of carefully establishing the optimal distance of grooves produced by discs acting on a confined surface, and the value (as a mere order of magnitude) of the increase of the thrust to produce the initiation of chip formation, as long as the confinement pressure becomes greater.

  16. E test as susceptibility test and epidemiologic tool for evaluation of Neisseria meningitidis isolates.

    PubMed Central

    Hughes, J H; Biedenbach, D J; Erwin, M E; Jones, R N

    1993-01-01

    The E test (AB Biodisk, Solna, Sweden), a new approach developed to test antimicrobial susceptibility, was compared with the agar dilution method for seven-drug antibiogram analysis of Neisseria meningitidis isolates. The overall E-test quantitative accuracy (+/- 1 log2 dilution) was 93% compared with that of agar dilution testing. The E test was then used to perform the susceptibility tests on a 10-year sample of 102 N. meningitidis isolates, including 5 from a recent epidemic outbreak in the University of Iowa (Iowa City) community. The E test proved to be an efficient methodology for identifying common source clusters of meningococcal disease having resistance to rifampin or sulfonamides. Moreover, the data demonstrated a recent increase in penicillin MICs (MIC for 90% of strains, 0.094 microgram/ml) and an escalation of high-level resistance to trimethoprimsulfamethoxazole (33%) and rifampin (14%). The E test should be considered a simple and accurate susceptibility method for the emerging need to test meningococci and other pathogenic neisserias. Chocolate Mueller-Hinton agar was observed to provide the best support of growth and E-test MIC results that correlated well with results of the reference agar dilution method previously used for neisserias. PMID:8308119

  17. High-Throughput Intracellular Antimicrobial Susceptibility Testing of Legionella pneumophila

    PubMed Central

    Chiaraviglio, Lucius

    2015-01-01

    Legionella pneumophila is a Gram-negative opportunistic human pathogen that causes a severe pneumonia known as Legionnaires' disease. Notably, in the human host, the organism is believed to replicate solely within an intracellular compartment, predominantly within pulmonary macrophages. Consequently, successful therapy is predicated on antimicrobials penetrating into this intracellular growth niche. However, standard antimicrobial susceptibility testing methods test solely for extracellular growth inhibition. Here, we make use of a high-throughput assay to characterize intracellular growth inhibition activity of known antimicrobials. For select antimicrobials, high-resolution dose-response analysis was then performed to characterize and compare activity levels in both macrophage infection and axenic growth assays. Results support the superiority of several classes of nonpolar antimicrobials in abrogating intracellular growth. Importantly, our assay results show excellent correlations with prior clinical observations of antimicrobial efficacy. Furthermore, we also show the applicability of high-throughput automation to two- and three-dimensional synergy testing. High-resolution isocontour isobolograms provide in vitro support for specific combination antimicrobial therapy. Taken together, findings suggest that high-throughput screening technology may be successfully applied to identify and characterize antimicrobials that target bacterial pathogens that make use of an intracellular growth niche. PMID:26392509

  18. High-Throughput Intracellular Antimicrobial Susceptibility Testing of Legionella pneumophila.

    PubMed

    Chiaraviglio, Lucius; Kirby, James E

    2015-12-01

    Legionella pneumophila is a Gram-negative opportunistic human pathogen that causes a severe pneumonia known as Legionnaires' disease. Notably, in the human host, the organism is believed to replicate solely within an intracellular compartment, predominantly within pulmonary macrophages. Consequently, successful therapy is predicated on antimicrobials penetrating into this intracellular growth niche. However, standard antimicrobial susceptibility testing methods test solely for extracellular growth inhibition. Here, we make use of a high-throughput assay to characterize intracellular growth inhibition activity of known antimicrobials. For select antimicrobials, high-resolution dose-response analysis was then performed to characterize and compare activity levels in both macrophage infection and axenic growth assays. Results support the superiority of several classes of nonpolar antimicrobials in abrogating intracellular growth. Importantly, our assay results show excellent correlations with prior clinical observations of antimicrobial efficacy. Furthermore, we also show the applicability of high-throughput automation to two- and three-dimensional synergy testing. High-resolution isocontour isobolograms provide in vitro support for specific combination antimicrobial therapy. Taken together, findings suggest that high-throughput screening technology may be successfully applied to identify and characterize antimicrobials that target bacterial pathogens that make use of an intracellular growth niche. PMID:26392509

  19. Antibacterial susceptibility of plaque bacteria.

    PubMed

    Newman, M G; Hulem, C; Colgate, J; Anselmo, C

    1979-07-01

    Selected anaerobic, capnophilic and facultative bacteria isolated from patients with various forms of periodontal health and disease were tested for their susceptibility to antibiotics and antimicrobial agents. Specific bactericidal and minimum inhibitory concentrations were compared to disc zone diameters, thereby generating new standards for the potential selection of antimicrobial agents. PMID:286720

  20. Rapid drug susceptibility test of mycobacterium tuberculosis by bioluminescence sensor

    NASA Astrophysics Data System (ADS)

    Lu, Bin; Xu, Shunqing; Chen, Zifei; Zhou, Yikai

    2001-09-01

    With the persisting increase of drug-resistant stains of M. Tuberculosis around the world, rapid and sensitive detection of antibiotic of M. Tuberculosis is becoming more and more important. In the present study, drug susceptibility of M. tuberculosis were detected by recombination mycobacteriophage combined with bioluminescence sensor. It is based on the use of recombination mycobacteriophage which can express firefly luciferase when it infects viable mycobacteria, and can effectively produce quantifiable photon. Meanwhile, in mycobacterium cells treated with active antibiotic, no light is observed. The emitted light is recorded by a bioluminscence sensor, so the result of drug-resistant test can be determined by the naked eye. 159 stains of M. tuberculosis were applied to this test on their resistant to rifampin, streptomycin and isoniazid. It is found that the agreement of this assay with Liewenstein- Jensen slat is: rifampin 95.60 percent, isoniazid 91.82 percent, streptomycin 88.68 percent, which showed that it is a fast and practical method to scene and detect drug resistant of mycobacterium stains.

  1. Rapid Bead-Based Antimicrobial Susceptibility Testing by Optical Diffusometry

    PubMed Central

    Chung, Chih-Yao; Wang, Jhih-Cheng; Chuang, Han-Sheng

    2016-01-01

    This study combined optical diffusometry and bead-based immunoassays to develop a novel technique for quantifying the growth of specific microorganisms and achieving rapid AST. Diffusivity rises when live bacteria attach to particles, resulting in additional energy from motile microorganisms. However, when UV-sterilized (dead) bacteria attach to particles, diffusivity declines. The experimental data are consistent with the theoretical model predicted according to the equivalent volume diameter. Using this diffusometric platform, the susceptibility of Pseudomonas aeruginosa to the antibiotic gentamicin was tested. The result suggests that the proliferation of bacteria is effectively controlled by gentamicin. This study demonstrated a sensitive (one bacterium on single particles) and time-saving (within 2 h) platform with a small sample volume (~0.5 μL) and a low initial bacteria count (50 CFU per droplet ~ 105 CFU/mL) for quantifying the growth of microorganisms depending on Brownian motion. The technique can be applied further to other bacterial strains and increase the success of treatments against infectious diseases in the near future. PMID:26863001

  2. Rapid Bead-Based Antimicrobial Susceptibility Testing by Optical Diffusometry.

    PubMed

    Chung, Chih-Yao; Wang, Jhih-Cheng; Chuang, Han-Sheng

    2016-01-01

    This study combined optical diffusometry and bead-based immunoassays to develop a novel technique for quantifying the growth of specific microorganisms and achieving rapid AST. Diffusivity rises when live bacteria attach to particles, resulting in additional energy from motile microorganisms. However, when UV-sterilized (dead) bacteria attach to particles, diffusivity declines. The experimental data are consistent with the theoretical model predicted according to the equivalent volume diameter. Using this diffusometric platform, the susceptibility of Pseudomonas aeruginosa to the antibiotic gentamicin was tested. The result suggests that the proliferation of bacteria is effectively controlled by gentamicin. This study demonstrated a sensitive (one bacterium on single particles) and time-saving (within 2 h) platform with a small sample volume (~0.5 μL) and a low initial bacteria count (50 CFU per droplet ~ 105 CFU/mL) for quantifying the growth of microorganisms depending on Brownian motion. The technique can be applied further to other bacterial strains and increase the success of treatments against infectious diseases in the near future. PMID:26863001

  3. Rifampin Heteroresistance in Mycobacterium tuberculosis Cultures as Detected by Phenotypic and Genotypic Drug Susceptibility Test Methods

    PubMed Central

    Folkvardsen, Dorte Bek; Thomsen, Vibeke Ø.; Rigouts, Leen; Rasmussen, Erik Michael; Bang, Didi; Bernaerts, Gertjan; Werngren, Jim; Toro, Juan Carlos; Hoffner, Sven; Hillemann, Doris

    2013-01-01

    Tuberculosis patients may harbor both drug-susceptible and -resistant bacteria, i.e., heteroresistance. We used mixtures of rifampin-resistant and -susceptible Mycobacterium tuberculosis strains to simulate heteroresistance in patient samples. Molecular tests can be used for earlier discovery of multidrug resistance (MDR), but the sensitivity to detect heteroresistance is unknown. Conventional phenotypic drug susceptibility testing was the most sensitive, whereas two line probe assays and sequencing were unable to detect the clinically important 1% resistant bacteria. PMID:24068005

  4. Testing the Susceptibility of GNSS Receivers to Radio Frequency Interference

    NASA Astrophysics Data System (ADS)

    Berglund, H. T.; Blume, F.; Gallaher, W. W.

    2015-12-01

    Global Navigational Satellite Systems (GNSS) receivers are employed by the scientific community for measuring a variety of geodetic, geophysical and atmospheric phenomena. Data acquisition frequently occurs in a variety of challenging environments, which include locations with high Radio Frequency (RF) noise characteristics. Tracking the relatively low powered GNSS carrier signals broadcast from space becomes even more challenging in the presence of adjacent band RF noise. The demand for terrestrial RF spectrum use for a variety of non-GNSS applications is ever increasing, which poses potential challenges for GNSS site operators who would like to acquire the highest quality data possible. In recent years, UNAVCO has observed an increase in the number of GNSS sites which are negatively impacted by RF interference. In previous work, we have shown that telemetry systems utilizing the Iridium satellite constellation can degrade GNSS data quality, as the adjacent-band (1610-1616 Mhz) signals transmitted by Iridium data transmitters are close in proximity to the L1 frequency of GNSS. The impact of RF interference from Iridium data transmitters on GNSS receivers can cause reduced Signal-to-Noise (SNR), increased cycle slips, and in worst case scenarios, prevent the receiver from tracking. To better characterize GNSS receiver susceptibility to RF interference, UNAVCO has performed a variety of tests with Continuous Wave (CW) noise sources in RF bands adjacent to the GNSS spectrum. We simulate a subset of discrete noise frequencies commonly observed in the field using a frequency generator, which supplies a signal with varying power output from a transmitter located within 1 m of the GNSS antenna. Signal power is incremented in small steps until receiver tracking fails. All receivers are simultaneously evaluated using an 8-way splitter. In addition, we investigate receiver tracking performance with a simulated increase in the RF noise floor. To analyze the results we use

  5. DKG statement on the use of metal alloy discs for patch testing in suspected intolerance to metal implants.

    PubMed

    Thomas, Peter; Geier, Johannes; Dickel, Heinrich; Diepgen, Thomas; Hillen, Uwe; Kreft, Burkhard; Schnuch, Axel; Szliska, Christiane; Mahler, Vera

    2015-10-01

    Intolerance reactions to metal implants may be caused by metal allergy. However, prior to implantation, 'prophetic'/prophylactic patch testing should not be performed. Pre-implant patch testing should only be done to verify or exclude metal allergy in patients with a corresponding history. In case of implant-related complications - in particular following replacement arthroplasty - such as pain, effusion, skin lesions, reduced range of motion or implant loosening, orthopedic causes should be ruled out first. Workup of suspected metal implant allergy should then be done using the DKG standard series, which includes nickel, cobalt, and chromium preparations. Various studies assessing the usefulness of metal alloy discs for patch testing have shown this particular approach to be ineffective with respect to providing reliable information on metal allergy. Any positive reaction in such tests cannot be assigned to a specific metal contained within the alloy. Furthermore, there is a risk of broad and indiscriminate use of these readily available discs. Accordingly, given the lack of additional benefit compared to patch testing with standardized metal salt preparations, we do not recommend patch testing with metal alloy discs. PMID:26408461

  6. The implications of endemic IMP-4 carbapenemase for clinical laboratory susceptibility testing.

    PubMed

    Goire, Namraj; Harnett, Gerald B; O'Reilly, Lyn C; Ingram, Paul R; Leung, Michael J; Speers, Davild J; Healy, Paul E; Inglis, Timothy J J

    2016-05-01

    A local predominance of carbapenemase producing Enterobacteriaceae with low minimum inhibitory concentrations (MIC) to meropenem prompted a review of methods available for carbapenemase detection. We report on results using two selective media, temocillin discs, CarbaNP test, GeneXpert Carba-R assay and an in-house PCR assay. PMID:26945518

  7. Survey of disc diffusion antimicrobial sensitivity testing in avian bacteriology laboratories and the evaluation of a standardised method.

    PubMed

    Whithear, K G; Htwe, T; Sulaiman, I

    1986-04-01

    A survey was conducted in which 15 laboratories involved in avian bacteriology tested the antimicrobial sensitivity of cultures of Staphylococcus aureus, S. epidermidis, Escherichia coli, Alcaligenes faecalis and Salmonella typhimurium, using the disc diffusion test as routinely practised in each laboratory. Up to 28 different antimicrobials or antimicrobial combinations were used. The most commonly tested agents were ampicillin, benzylpenicillin, chloramphenicol, erythromycin, furazolidone, neomycin, streptomycin, sulphonamide, tetracycline and trimethoprim/sulphonamide. Results between laboratories were compared according to the width of the zone of inhibition (annular radius) where 5 or more laboratories used the same disc content of a particular antimicrobial agent, and on whether a culture was interpreted as being sensitive or resistant to a particular agent. The variation in annular radii about the mean values were less than +/- 2 SD in 39.5% and greater than +/- 3 SD in 32.6% of 43 different antimicrobial/culture combinations involving 8 different agents. The range of annular radii measurements was greatest with trimethoprim/sulphonamide and streptomycin discs and with the S. epidermidis culture. Variation in the interpretation of test results was greatest with the Gram-negative bacteria, where for each of the more frequently tested agents at least one, commonly 2 and sometimes all 3 cultures were reported as sensitive and resistant to the same agent by different laboratories. The calibrated dichotomous sensitivity (CDS) test of Bell (1975, 1984) was evaluated as a standardised disc diffusion procedure and was found to be reproducible and accurate. Results obtained using antimicrobial agents calibrated for use in the CDS test were compared with results obtained in the survey using the same agents or agents showing identical resistance patterns.(ABSTRACT TRUNCATED AT 250 WORDS) PMID:3741274

  8. A freeze-thaw test to determine the frost susceptibility of soils

    NASA Astrophysics Data System (ADS)

    Chamberlain, Edwin J.

    1987-01-01

    A freezing test for determining the frost susceptibility of soils is presented to supplant the standard CRREL freezing test currently specified by the Corps of Engineers. This test reduces the time required to determine the frost susceptibility of a soil in half. It also allows for the determination of both the frost heave and thaw weakening susceptibilities and considers the effects of the freeze-thaw cycling. The freeze test eliminates much of the variability in test results caused by the human element by completely automating the temperature control and data observations.

  9. Preparation of ormetoprim sulfadimethoxine medicated discs for disc diffusion assay

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Romet (a blend of ormetoprim and sulfadimethoxine) is a typeA medicated article for the manufacture of medicated feed in the catfish industry. Recently, the commercial manufacture of ormetoprim–sulfadimethoxine susceptibility discs was discontinued. Ormetoprim–sulfadimethoxine discs were prepared at...

  10. Testing the molecular-hydrogen Kennicutt-Schmidt law in the low-density environments of extended ultraviolet disc galaxies

    NASA Astrophysics Data System (ADS)

    Watson, Linda C.; Martini, Paul; Lisenfeld, Ute; Böker, Torsten; Schinnerer, Eva

    2016-01-01

    Studying star formation beyond the optical radius of galaxies allows us to test empirical relations in extreme conditions with low average gas density and low molecular fraction. Previous studies discovered galaxies with extended ultraviolet (XUV) discs, which often contain star-forming regions with lower Hα-to-far-UV (FUV) flux ratios compared to inner disc star-forming regions. However, most previous studies lack measurements of molecular gas, which is presumably the component of the interstellar medium out of which stars form. We analysed published CO measurements and upper limits for 15 star-forming regions in the XUV or outer disc of three nearby spiral galaxies and a new CO upper limit from the IRAM (Institut de Radioastronomie Millimétrique) 30 m telescope in one star-forming region at r = 3.4r25 in the XUV disc of NGC 4625. We found that the star-forming regions are in general consistent with the same molecular-hydrogen Kennicutt-Schmidt law that applies within the optical radius, independent of whether we used Hα or FUV as the star formation rate (SFR) tracer. However, a number of the CO detections are significantly offset towards higher SFR surface density for their molecular-hydrogen surface density. Deeper CO data may enable us to use the presence or absence of molecular gas as an evolutionary probe to break the degeneracy between age and stochastic sampling of the initial mass function as the explanation for the low Hα-to-FUV flux ratios in XUV discs.

  11. Susceptibility testing of Danish isolates of Capnocytophaga and CDC group DF-2 bacteria.

    PubMed

    Bremmelgaard, A; Pers, C; Kristiansen, J E; Korner, B; Heltberg, O; Frederiksen, W

    1989-01-01

    Twelve Capnocytophaga and seven DF-2 strains were tested for their susceptibility to 14 antimicrobial agents using an agar dilution and an agar diffusion method. Twenty-three other antibiotics were evaluated using the diffusion test only. All strains were fully susceptible to penicillin, ampicillin, cefuroxime, cefotaxime, erythromycin, clindamycin, chloramphenicol, doxycycline, rifamycin and ofloxacin using both methods. Clindamycin, rifamycin and cefotaxime were most active. Using agar dilution some strains were susceptible to gentamicin, but agar diffusion showed total resistance. One Capnocytophaga strain was susceptible and another moderately susceptible to metronidazole, other strains were resistant. The agar diffusion test showed that both Capnocytophaga and DF-2 were resistant to most other aminoglycosides, to fosfomycin, polymyxin and trimethoprim. All strains of both taxa were fully susceptible to piperacillin, cefoxitin, imipenem and fusidic acid and showed different susceptibilities to the other agents. Susceptibility testing by means of agar diffusion using an enriched chocolate agar and 5% CO2 atmosphere could be used to test Capnocytophaga and DF-2 strains and gives sufficient accuracy for routine use, when revised inhibition zone breakpoints are employed. PMID:2914105

  12. Antimicrobial susceptibility testing of Spanish field isolates of Brachyspira hyodysenteriae.

    PubMed

    Hidalgo, A; Carvajal, A; García-Feliz, C; Osorio, J; Rubio, P

    2009-08-01

    This study is the first conducted in Spain to evaluate antimicrobial susceptibility of field isolates of Brachyspira hyodysenteriae. One hundred and eight isolates of the bacterium, recovered from different Spanish swine farms between 2000 and 2007, were investigated. The minimum inhibitory concentrations (MIC) of erythromycin, tylosin, tiamulin, valnemulin, clindamycin and lincomycin were determined using a broth microdilution technique. Most of the isolates showed poor susceptibility to erythromycin (MIC(90)>256 microg/ml), tylosin (MIC(90)>256 microg/ml), clindamycin (MIC(90)>4 microg/ml) and lincomycin (MIC(90)=128 microg/ml). Reduced susceptibility to tiamulin and valnemulin was observed with a MIC>2 microg/ml in 17.6% and 7.41% of the B. hyodysenteriae isolates, respectively. Moreover, a survival analysis permitted the detection of an increasing trend in the MIC values for almost all the antimicrobials used in the treatment of swine dysentery when comparing recent isolates (from 2006 to 2007) with those recovered in earlier years (between 2000 and 2004). PMID:19084246

  13. A Novel Hypoxia Challenge Test Demonstrates Cardiovascular and Pulmonary Susceptibility to Acrolein Gas in Hypertensive Rats.

    EPA Science Inventory

    High levels of air pollution increase the risk of cardiovascular morbidity and mortality, especially in susceptible populations including those with hypertension. Stress tests are useful for manifesting latent effects of exposure, particularly at low concentrations, often when no...

  14. Susceptibility Testing of Anaerobic Bacteria with 100-μg Carbenicillin Disks

    PubMed Central

    Laslie, W. W.; Lambe, D. W.

    1976-01-01

    A total of 245 strains of anaerobic bacteria were examined for their susceptibility to carbenicillin by the disk test method and by minimum inhibitory concentration (MIC) determinations. Standard-curve studies with a strain of Bacteroides fragilis subsp. fragilis that was minimally susceptible to carbenicillin and Escherichia coli (ATCC 25922) demonstrated that a disk containing 100 μg of carbenicillin was suitable for testing susceptibility of anaerobes to carbenicillin. Thus, the diameter of zones around the 100-μg carbenicillin disks and MIC values were determined under the following test conditions: Mueller-Hinton agar supplemented with sheep blood, vitamin K1, and hemin; an incubation temperature of 35 C; and an atmosphere of 80% N2, 10% H2, and 10% CO2. The strains were separated into two populations by correlating zone diameters and geometric mean MICs. The disk test more clearly separated the resistant and susceptible populations and was more reproducible than the MIC test. Thus, a statistical analysis based on the distribution of zone diameters of susceptible and resistant strains was used to derive an interpretive scheme for anaerobic bacteria tested with 100-μg carbenicillin disks. The following interpretive scheme is recommended for testing anaerobes with 100-μg disks of carbenicillin: resistant, 8 mm or less; indeterminate, 9 to 12 mm; and susceptible, 13 mm or greater. PMID:984743

  15. Antimicrobial susceptibility testing for bovine respiratory disease: getting more from diagnostic results.

    PubMed

    Lubbers, Brian V; Turnidge, John

    2015-02-01

    Bovine respiratory disease (BRD) is one of the most common diseases of cattle worldwide. Given the significant bacterial component of this disease, antimicrobial agents remain one of the mainstays of therapy. However, the potential welfare and economic impact resulting from the selection of inappropriate antimicrobial therapy for BRD poses significant risks to both animal and animal owner. To determine the 'best' antimicrobial agent for a specific case, the decision-making process needs to incorporate all available evidence, often including the results of bacterial culture and antimicrobial susceptibility testing. While antimicrobial susceptibility testing can be a valuable diagnostic tool, integrating the test results into the clinical decision making process can be a challenging experience. This review details the process by which interpretive criteria for susceptibility tests are developed. Principles for how to best integrate antimicrobial susceptibility testing, both at the individual animal test and aggregate test levels, into the clinical decision making process are discussed. Non-traditional testing methodologies and how they may improve susceptibility testing in the future are also reviewed. PMID:25582794

  16. Bacterial Cytological Profiling (BCP) as a Rapid and Accurate Antimicrobial Susceptibility Testing Method for Staphylococcus aureus

    PubMed Central

    Quach, D.T.; Sakoulas, G.; Nizet, V.; Pogliano, J.; Pogliano, K.

    2016-01-01

    Successful treatment of bacterial infections requires the timely administration of appropriate antimicrobial therapy. The failure to initiate the correct therapy in a timely fashion results in poor clinical outcomes, longer hospital stays, and higher medical costs. Current approaches to antibiotic susceptibility testing of cultured pathogens have key limitations ranging from long run times to dependence on prior knowledge of genetic mechanisms of resistance. We have developed a rapid antimicrobial susceptibility assay for Staphylococcus aureus based on bacterial cytological profiling (BCP), which uses quantitative fluorescence microscopy to measure antibiotic induced changes in cellular architecture. BCP discriminated between methicillin-susceptible (MSSA) and -resistant (MRSA) clinical isolates of S. aureus (n = 71) within 1–2 h with 100% accuracy. Similarly, BCP correctly distinguished daptomycin susceptible (DS) from daptomycin non-susceptible (DNS) S. aureus strains (n = 20) within 30 min. Among MRSA isolates, BCP further identified two classes of strains that differ in their susceptibility to specific combinations of beta-lactam antibiotics. BCP provides a rapid and flexible alternative to gene-based susceptibility testing methods for S. aureus, and should be readily adaptable to different antibiotics and bacterial species as new mechanisms of resistance or multidrug-resistant pathogens evolve and appear in mainstream clinical practice. PMID:26981574

  17. Bacterial Cytological Profiling (BCP) as a Rapid and Accurate Antimicrobial Susceptibility Testing Method for Staphylococcus aureus.

    PubMed

    Quach, D T; Sakoulas, G; Nizet, V; Pogliano, J; Pogliano, K

    2016-02-01

    Successful treatment of bacterial infections requires the timely administration of appropriate antimicrobial therapy. The failure to initiate the correct therapy in a timely fashion results in poor clinical outcomes, longer hospital stays, and higher medical costs. Current approaches to antibiotic susceptibility testing of cultured pathogens have key limitations ranging from long run times to dependence on prior knowledge of genetic mechanisms of resistance. We have developed a rapid antimicrobial susceptibility assay for Staphylococcus aureus based on bacterial cytological profiling (BCP), which uses quantitative fluorescence microscopy to measure antibiotic induced changes in cellular architecture. BCP discriminated between methicillin-susceptible (MSSA) and -resistant (MRSA) clinical isolates of S. aureus (n = 71) within 1-2 h with 100% accuracy. Similarly, BCP correctly distinguished daptomycin susceptible (DS) from daptomycin non-susceptible (DNS) S. aureus strains (n = 20) within 30 min. Among MRSA isolates, BCP further identified two classes of strains that differ in their susceptibility to specific combinations of beta-lactam antibiotics. BCP provides a rapid and flexible alternative to gene-based susceptibility testing methods for S. aureus, and should be readily adaptable to different antibiotics and bacterial species as new mechanisms of resistance or multidrug-resistant pathogens evolve and appear in mainstream clinical practice. PMID:26981574

  18. Perceived susceptibility and self-protective behavior: a field experiment to encourage home radon testing

    SciTech Connect

    Weinstein, N.D.; Sandman, P.M.; Roberts, N.E. )

    1991-01-01

    Tested in a field experiment (N = 647) the hypothesis that perceptions of personal susceptibility are important in decisions to test one's home for radioactive radon gas. Experimental group subjects received a personal telephone call to tell them they lived in a high-risk area and a personal letter to reinforce the telephone message. After the intervention, experimental subjects were significantly more likely than minimal-treatment subjects to acknowledge the possibility of high radon levels in their homes. Perceptions of susceptibility and illness severity were significantly correlated with orders of radon test kits and with testing intentions. Nevertheless, there were no differences between groups in test orders or intentions. Results are discussed in terms of the difficulty of getting people to acknowledge susceptibility and the factors other than risk perceptions that influence self-protective behavior.

  19. Antimicrobial susceptibility testing in 90 min by bacterial cell count monitoring

    PubMed Central

    Broeren, M A C; Maas, Y; Retera, E; Arents, N L A

    2013-01-01

    The rise in antimicrobial resistance has become a serious global health problem. Restrictive use of antibiotics seems the only option to temper this accession since research in new antibiotics has halted. Antimicrobial stewardship programmes rely on quick access to susceptibility data. This study evaluated the concept of bacterial cell count monitoring as a fast method to determine susceptibility. Escherichia coli, Pseudomonas aeruginosa and Staphylococcus aureus strains were tested for amoxicillin/piperacillin and gentamicin by three conventional methods (VITEK2®, Etest® and broth-macrodilution). Bacterial cell count monitoring reliably predicted susceptibility after 90 min for Escherichia coli and after 120 min for Pseudomonas aeruginosa and Staphylococcus aureus without any minor, major or very major discrepancies. Time-to-result was reduced by 74%, 83% and 76%, respectively. Bacterial cell count monitoring shows great potential for rapid susceptibility testing. PMID:22390723

  20. Antimicrobial susceptibility testing in 90 min by bacterial cell count monitoring.

    PubMed

    Broeren, M A C; Maas, Y; Retera, E; Arents, N L A

    2013-03-01

    The rise in antimicrobial resistance has become a serious global health problem. Restrictive use of antibiotics seems the only option to temper this accession since research in new antibiotics has halted. Antimicrobial stewardship programmes rely on quick access to susceptibility data. This study evaluated the concept of bacterial cell count monitoring as a fast method to determine susceptibility. Escherichia coli, Pseudomonas aeruginosa and Staphylococcus aureus strains were tested for amoxicillin/piperacillin and gentamicin by three conventional methods (VITEK2(®) , Etest(®) and broth-macrodilution). Bacterial cell count monitoring reliably predicted susceptibility after 90 min for Escherichia coli and after 120 min for Pseudomonas aeruginosa and Staphylococcus aureus without any minor, major or very major discrepancies. Time-to-result was reduced by 74%, 83% and 76%, respectively. Bacterial cell count monitoring shows great potential for rapid susceptibility testing. PMID:22390723

  1. Genetic susceptibility testing from a stress and coping perspective.

    PubMed

    Gooding, Holly C; Organista, Kurt; Burack, Jeffrey; Biesecker, Barbara Bowles

    2006-04-01

    Four theories of health behavior and of stress and coping are reviewed for their ability to illuminate interest in uptake and outcomes of genetic testing for adult-onset diseases. These theories are the Health Belief Model, the Theory of Planned Behavior (TPB), the Common Sense Model of Self-regulation (CSM), and the Transactional Model of Stress and Coping (TMSC). Basic concepts of each theory are discussed, followed by evidence from the literature supporting the relevance of these concepts to the understanding of genetic testing for four adult-onset diseases: Huntington's disease, Alzheimer's disease, hereditary breast/ovarian cancer, and hereditary colorectal cancer. Emphasis is placed on the finding that a decision to undergo genetic testing may be considered as a way to cope with both the cognitive and affective concerns that arise from living at increased risk of developing a disease in the future. The potential value of genetic testing for reducing uncertainty about and gaining a sense of control over one's risk of developing a chronic disease is highlighted. We argue that theories which focus on stress and coping provide a useful framework for future studies of genetic testing decisions for adult-onset disease risk. PMID:16198036

  2. Identification and susceptibility testing of Staphylococcus aureus by direct inoculation from positive BACTEC blood culture bottles.

    PubMed

    Diederen, B M W; Zieltjens, M; Wetten, H; Buiting, A G M

    2006-01-01

    This study explored the possibility of combining direct inoculation of tube coagulase and DNase tests, and the VITEK2 system, from BACTEC blood culture bottles in order to achieve rapid identification and susceptibility testing of Staphylococcus aureus. All isolates were identified correctly as S. aureus or coagulase-negative staphylococci (CNS). Antimicrobial susceptibility testing with the VITEK2 system gave 99.6% correct category agreement, with 0.1% very major errors and 0.3% minor errors among S. aureus isolates, and 97.4% correct category agreement, with 0.9% very major errors and 1.7% minor errors among CNS isolates. The results suggested that direct identification and susceptibility testing is sufficiently accurate for immediate reporting. PMID:16460552

  3. Effects of follower load and rib cage on intervertebral disc pressure and sagittal plane curvature in static tests of cadaveric thoracic spines.

    PubMed

    Anderson, Dennis E; Mannen, Erin M; Sis, Hadley L; Wong, Benjamin M; Cadel, Eileen S; Friis, Elizabeth A; Bouxsein, Mary L

    2016-05-01

    The clinical relevance of mechanical testing studies of cadaveric human thoracic spines could be enhanced by using follower preload techniques, by including the intact rib cage, and by measuring thoracic intervertebral disc pressures, but studies to date have not incorporated all of these components simultaneously. Thus, this study aimed to implement a follower preload in the thoracic spine with intact rib cage, and examine the effects of follower load, rib cage stiffening and rib cage removal on intervertebral disc pressures and sagittal plane curvatures in unconstrained static conditions. Intervertebral disc pressures increased linearly with follower load magnitude. The effect of the rib cage on disc pressures in static conditions remains unclear because testing order likely confounded the results. Disc pressures compared well with previous reports in vitro, and comparison with in vivo values suggests the use of a follower load of about 400N to approximate loading in upright standing. Follower load had no effect on sagittal plane spine curvature overall, suggesting successful application of the technique, although increased flexion in the upper spine and reduced flexion in the lower spine suggest that the follower load path was not optimized. Rib cage stiffening and removal both increased overall spine flexion slightly, although with differing effects at specific spinal locations. Overall, the approaches demonstrated here will support the use of follower preloads, intact rib cage, and disc pressure measurements to enhance the clinical relevance of future studies of the thoracic spine. PMID:26944690

  4. Testing candidate genes that may affect susceptibility to leprosy.

    PubMed

    Cervino, A C; Curnow, R N

    1997-12-01

    Several statistical methods have been used to search familial data sets for marker alleles associated with the occurrence of a disease. In the present paper, a recently developed method is used to re-analyze published data on leprosy and candidate genes at the HLA loci. This new method of analysis, the randomization transmission disequilibrium test (TDT), confirmed previous conclusions that there was no significant evidence against random transmission at the HLA-A locus but significant positive association with the HLA-DR2 allele. The randomization TDT detected significant protective associations, that had not previously been found, with alleles HLA-B8 in Egyptian families and HLA-B21 (current nomenclature B x 4901, 5001-5002) in South Indian families, highlighting a major advantage of permutation tests in analyzing candidate gene loci with rare alleles. These findings provide evidence that HLA class I restricted T lymphocytes may be of protective importance in leprosy. PMID:9465154

  5. A Simplified Test for Blanching Susceptibility of Copper Alloys

    NASA Technical Reports Server (NTRS)

    Thomas-Ogbuji, Linus U.; Humphrey, Donald; Setlock, John

    2003-01-01

    GRCop-84 (Cu-8Cr-4Nb) is a dispersion-strengthened alloy developed for space-launch rocket engine applications, as a liner for the combustion chamber and nozzle ramp. Its main advantage over rival alloys, particularly NARloy-Z (Cu-Ag-Zr), the current liner alloy, is in high temperature mechanical properties. Further validation required that the two alloys be compared with respect to service performance and durability. This has been done, under conditions resembling those expected in reusable launch engine applications. GRCop-84 was found to have a superior resistance to static and cyclic oxidation up to approx. 700 C. In order to improve its performance above 700 C, Cu-Cr coatings have also been developed and evaluated. The major oxidative issue with Cu alloys is blanching, a mode of degradation induced by oxidation-reduction fluctuations in hydrogen-fueled engines. That fluctuation cannot be addressed with conventional static or cyclic oxidation testing. Hence, a further evaluation of the alloy substrates and Cu-Cr coating material necessitated our devising a test protocol that involves oxidaton-reduction cycles. This paper describes the test protocols used and the results obtained.

  6. A Meta-Analytic Investigation of the Susceptibility of Integrity Tests to Faking and Coaching.

    ERIC Educational Resources Information Center

    Alliger, George M.; Dwight, Stephen A.

    2000-01-01

    Performed a meta-analysis of studies of the extent to which individuals can inflate their integrity test scores when coached or instructed to fake good. Results from 14 studies indicate that overt tests are more susceptible to both fake-good and coaching instructions than are personality-based measures. (SLD)

  7. A two-hour antibiotic susceptibility test by ATP-bioluminescence.

    PubMed

    March Rosselló, Gabriel Alberto; García-Loygorri Jordán de Urries, María Cristina; Gutiérrez Rodríguez, María Purificación; Simarro Grande, María; Orduña Domingo, Antonio; Bratos Pérez, Miguel Ángel

    2016-01-01

    The antibiotic susceptibility test (AST) in Clinical Microbiology laboratories is still time-consuming, and most procedures take 24h to yield results. In this study, a rapid antimicrobial susceptibility test using ATP-bioluminescence has been developed. The design of method was performed using five ATCC collection strains of known susceptibility. This procedure was then validated against standard commercial methods on 10 strains of enterococci, 10 staphylococci, 10 non-fermenting gram negative bacilli, and 13 Enterobacteriaceae from patients. The agreement obtained in the sensitivity between the ATP-bioluminescence method and commercial methods (E-test, MicroScan and VITEK2) was 100%. In summary, the preliminary results obtained in this work show that the ATP-bioluminescence method could provide a fast and reliable AST in two hours. PMID:25979598

  8. Comparison of agar dilution and antibiotic gradient strip test with broth microdilution for susceptibility testing of swine Brachyspira species.

    PubMed

    Mirajkar, Nandita S; Gebhart, Connie J

    2016-03-01

    Production-limiting diseases in swine caused by Brachyspira are characterized by mucohemorrhagic diarrhea (B. hyodysenteriae and "B. hampsonii") or mild colitis (B. pilosicoli), while B. murdochii is often isolated from healthy pigs. Emergence of novel pathogenic Brachyspira species and strains with reduced susceptibility to commonly used antimicrobials has reinforced the need for standardized susceptibility testing. Two methods are currently used for Brachyspira susceptibility testing: agar dilution (AD) and broth microdilution (BMD). However, these tests have primarily been used for B. hyodysenteriae and rarely for B. pilosicoli. Information on the use of commercial susceptibility testing products such as antibiotic gradient strips is lacking. Our main objective was to validate and compare the susceptibility results, measured as the minimum inhibitory concentration (MIC), of 6 antimicrobials for 4 Brachyspira species (B. hyodysenteriae, "B. hampsonii", B. pilosicoli, and B. murdochii) by BMD and AD (tiamulin, valnemulin, lincomycin, tylosin, and carbadox) or antibiotic gradient strip (doxycycline) methods. In general, the results of a high percentage of all 4 Brachyspira species differed by ±1 log2 dilution or less by BMD and AD for tiamulin, valnemulin, lincomycin, and tylosin, and by BMD and antibiotic gradient strip for doxycycline. The carbadox MICs obtained by BMD were 1-5 doubling dilutions different than those obtained by AD. BMD for Brachyspira was quicker to perform with less ambiguous interpretation of results when compared with AD and antibiotic gradient strip methods, and the results confirm the utility of BMD in routine diagnostics. PMID:26965233

  9. A Microfluidic Channel Method for Rapid Drug-Susceptibility Testing of Pseudomonas aeruginosa

    PubMed Central

    Matsumoto, Yoshimi; Grushnikov, Andrey; Kikuchi, Kazuma; Noji, Hiroyuki; Yamaguchi, Akihito; Yagi, Yasushi

    2016-01-01

    The recent global increase in the prevalence of antibiotic-resistant bacteria and lack of development of new therapeutic agents emphasize the importance of selecting appropriate antimicrobials for the treatment of infections. However, to date, the development of completely accelerated drug susceptibility testing methods has not been achieved despite the availability of a rapid identification method. We proposed an innovative rapid method for drug susceptibility testing for Pseudomonas aeruginosa that provides results within 3 h. The drug susceptibility testing microfluidic (DSTM) device was prepared using soft lithography. It consisted of five sets of four microfluidic channels sharing one inlet slot, and the four channels are gathered in a small area, permitting simultaneous microscopic observation. Antimicrobials were pre-introduced into each channel and dried before use. Bacterial suspensions in cation-adjusted Mueller–Hinton broth were introduced from the inlet slot and incubated for 3 h. Susceptibilities were microscopically evaluated on the basis of differences in cell numbers and shapes between drug-treated and control cells, using dedicated software. The results of 101 clinically isolated strains of P. aeruginosa obtained using the DSTM method strongly correlated with results obtained using the ordinary microbroth dilution method. Ciprofloxacin, meropenem, ceftazidime, and piperacillin caused elongation in susceptible cells, while meropenem also induced spheroplast and bulge formation. Morphological observation could alternatively be used to determine the susceptibility of P. aeruginosa to these drugs, although amikacin had little effect on cell shape. The rapid determination of bacterial drug susceptibility using the DSTM method could also be applicable to other pathogenic species, and it could easily be introduced into clinical laboratories without the need for expensive instrumentation. PMID:26872134

  10. A Microfluidic Channel Method for Rapid Drug-Susceptibility Testing of Pseudomonas aeruginosa.

    PubMed

    Matsumoto, Yoshimi; Sakakihara, Shouichi; Grushnikov, Andrey; Kikuchi, Kazuma; Noji, Hiroyuki; Yamaguchi, Akihito; Iino, Ryota; Yagi, Yasushi; Nishino, Kunihiko

    2016-01-01

    The recent global increase in the prevalence of antibiotic-resistant bacteria and lack of development of new therapeutic agents emphasize the importance of selecting appropriate antimicrobials for the treatment of infections. However, to date, the development of completely accelerated drug susceptibility testing methods has not been achieved despite the availability of a rapid identification method. We proposed an innovative rapid method for drug susceptibility testing for Pseudomonas aeruginosa that provides results within 3 h. The drug susceptibility testing microfluidic (DSTM) device was prepared using soft lithography. It consisted of five sets of four microfluidic channels sharing one inlet slot, and the four channels are gathered in a small area, permitting simultaneous microscopic observation. Antimicrobials were pre-introduced into each channel and dried before use. Bacterial suspensions in cation-adjusted Mueller-Hinton broth were introduced from the inlet slot and incubated for 3 h. Susceptibilities were microscopically evaluated on the basis of differences in cell numbers and shapes between drug-treated and control cells, using dedicated software. The results of 101 clinically isolated strains of P. aeruginosa obtained using the DSTM method strongly correlated with results obtained using the ordinary microbroth dilution method. Ciprofloxacin, meropenem, ceftazidime, and piperacillin caused elongation in susceptible cells, while meropenem also induced spheroplast and bulge formation. Morphological observation could alternatively be used to determine the susceptibility of P. aeruginosa to these drugs, although amikacin had little effect on cell shape. The rapid determination of bacterial drug susceptibility using the DSTM method could also be applicable to other pathogenic species, and it could easily be introduced into clinical laboratories without the need for expensive instrumentation. PMID:26872134

  11. Antibiotic Susceptibility Testing of the Gram-Negative Bacteria Based on Flow Cytometry

    PubMed Central

    Saint-Ruf, Claude; Crussard, Steve; Franceschi, Christine; Orenga, Sylvain; Ouattara, Jasmine; Ramjeet, Mahendrasingh; Surre, Jérémy; Matic, Ivan

    2016-01-01

    Rapidly treating infections with adequate antibiotics is of major importance. This requires a fast and accurate determination of the antibiotic susceptibility of bacterial pathogens. The most frequently used methods are slow because they are based on the measurement of growth inhibition. Faster methods, such as PCR-based detection of determinants of antibiotic resistance, do not always provide relevant information on susceptibility, particularly that which is not genetically based. Consequently, new methods, such as the detection of changes in bacterial physiology caused by antibiotics using flow cytometry and fluorescent viability markers, are being explored. In this study, we assessed whether Alexa Fluor® 633 Hydrazide (AFH), which targets carbonyl groups, can be used for antibiotic susceptibility testing. Carbonylation of cellular macromolecules, which increases in antibiotic-treated cells, is a particularly appropriate to assess for this purpose because it is irreversible. We tested the susceptibility of clinical isolates of Gram-negative bacteria, Escherichia coli and Pseudomonas aeruginosa, to antibiotics from the three classes: β-lactams, aminoglycosides, and fluoroquinolones. In addition to AFH, we used TO-PRO®-3, which enters cells with damaged membranes and binds to DNA, and DiBAC4 (3), which enters cells with depolarized membranes. We also monitored antibiotic-induced morphological alterations of bacterial cells by analyzing light scattering signals. Although all tested dyes and light scattering signals allowed for the detection of antibiotic-sensitive cells, AFH proved to be the most suitable for the fast and reliable detection of antibiotic susceptibility. PMID:27507962

  12. Antibiotic Susceptibility Testing of the Gram-Negative Bacteria Based on Flow Cytometry.

    PubMed

    Saint-Ruf, Claude; Crussard, Steve; Franceschi, Christine; Orenga, Sylvain; Ouattara, Jasmine; Ramjeet, Mahendrasingh; Surre, Jérémy; Matic, Ivan

    2016-01-01

    Rapidly treating infections with adequate antibiotics is of major importance. This requires a fast and accurate determination of the antibiotic susceptibility of bacterial pathogens. The most frequently used methods are slow because they are based on the measurement of growth inhibition. Faster methods, such as PCR-based detection of determinants of antibiotic resistance, do not always provide relevant information on susceptibility, particularly that which is not genetically based. Consequently, new methods, such as the detection of changes in bacterial physiology caused by antibiotics using flow cytometry and fluorescent viability markers, are being explored. In this study, we assessed whether Alexa Fluor® 633 Hydrazide (AFH), which targets carbonyl groups, can be used for antibiotic susceptibility testing. Carbonylation of cellular macromolecules, which increases in antibiotic-treated cells, is a particularly appropriate to assess for this purpose because it is irreversible. We tested the susceptibility of clinical isolates of Gram-negative bacteria, Escherichia coli and Pseudomonas aeruginosa, to antibiotics from the three classes: β-lactams, aminoglycosides, and fluoroquinolones. In addition to AFH, we used TO-PRO®-3, which enters cells with damaged membranes and binds to DNA, and DiBAC4 (3), which enters cells with depolarized membranes. We also monitored antibiotic-induced morphological alterations of bacterial cells by analyzing light scattering signals. Although all tested dyes and light scattering signals allowed for the detection of antibiotic-sensitive cells, AFH proved to be the most suitable for the fast and reliable detection of antibiotic susceptibility. PMID:27507962

  13. Clinical impact of rapid in vitro susceptibility testing and bacterial identification.

    PubMed Central

    Doern, G V; Vautour, R; Gaudet, M; Levy, B

    1994-01-01

    During the past decade, a variety of instrument-assisted bacterial identification and antimicrobial susceptibility test systems have been developed which permit provision of test results in a matter of hours rather than days, as has been the case with traditional overnight procedures. These newer rapid techniques are much more expensive than older methods. It has been presumed but not proven that the clinical benefits of rapid testing to patients with infection offset the added cost. The intent of this study was to objectively define the clinical impact of rapid bacterial identification and antimicrobial susceptibility testing. A 1-year study was performed in which infected, hospitalized patients in a tertiary-care, teaching, medical center were randomly assigned to one of two groups: patients for whom identification and susceptibility testing was performed by using a semi-automated, rapid, same-day procedure and those for whom testing was accomplished by using traditional overnight techniques. The two groups were compared with respect to numerous demographic descriptors, and then patients were monitored prospectively through the end of their hospitalization with the aim of determining whether there existed objectively defineable differences in management and outcome between the two groups. The mean lengths of time to provision of susceptibility and identification test results in the rapid test group were 11.3 and 9.6 h, respectively. In the overnight test group, these values were 19.6 and 25.9 h, respectively (P < 0.0005). There were 273 evaluable patients in the first group and 300 in the second group. Other than the length of time required to provide susceptibility and identification test results, no significant differences were noted between the two groups with respect to > 100 demographic descriptors. With regard to measures of outcome, the mean lengths of hospitalization were also the same in both groups. Mortality rates were however, lower in the rapid test

  14. Recommendation for a Standardised Method of Broth Microdilution Susceptibility Testing for Porcine Bordetella bronchiseptica

    PubMed Central

    Prüller, Sandra; Frömke, Cornelia; Kaspar, Heike; Klein, Günter; Kreienbrock, Lothar; Kehrenberg, Corinna

    2015-01-01

    The objective was to establish and standardise a broth microdilution susceptibility testing method for porcine Bordetella (B.) bronchiseptica. B. bronchiseptica isolates from different geographical regions and farms were genotyped by macrorestriction analysis and subsequent pulsed-field gel electrophoresis. One reference and one type strain plus two field isolates of B. bronchiseptica were chosen to analyse growth curves in four different media: cation-adjusted Mueller-Hinton broth (CAMHB) with and without 2% lysed horse blood, Brain-Heart-Infusion (BHI), and Caso broth. The growth rate of each test strain in each medium was determined by culture enumeration and the suitability of CAMHB was confirmed by comparative statistical analysis. Thereafter, reference and type strain and eight epidemiologically unrelated field isolates of B. bronchiseptica were used to test the suitability of a broth microdilution susceptibility testing method following CLSI-approved performance standards given in document VET01-A4. Susceptibility tests, using 20 antimicrobial agents, were performed in five replicates, and data were collected after 20 and 24 hours incubation and statistically analysed. Due to the low growth rate of B. bronchiseptica, an incubation time of 24 hours resulted in significantly more homogeneous minimum inhibitory concentrations after five replications compared to a 20-hour incubation. An interlaboratory comparison trial including susceptibility testing of 24 antimicrobial agents revealed a high mean level of reproducibility (97.9%) of the modified method. Hence, in a harmonization for broth microdilution susceptibility testing of B. bronchiseptica, an incubation time of 24 hours in CAMHB medium with an incubation temperature of 35°C and an inoculum concentration of approximately 5 x 105 cfu/ml was proposed. PMID:25910232

  15. Negative temperament as a moderator of intervention effects in infancy: testing a differential susceptibility model.

    PubMed

    Anzman-Frasca, Stephanie; Stifter, Cynthia A; Paul, Ian M; Birch, Leann L

    2014-10-01

    A consideration of potential moderators can highlight intervention effects that are attenuated when investigating aggregate results. Differential susceptibility is one type of interaction, where susceptible individuals have poorer outcomes in negative environments and better outcomes in positive environments, compared to less susceptible individuals, who have moderate outcomes regardless of environment. In the current study, we provide rationale for investigating this type of interaction in the context of a behavioral childhood obesity preventive intervention and test whether infant negativity moderated intervention effects on infant self-regulation and weight gain and on two aspects of mothers' parenting competence: parenting self-efficacy and parenting satisfaction. Results showed that infants' negative temperament at 3 weeks moderated intervention effects on some, but not all, outcomes. The intervention led to greater parenting satisfaction in mothers with highly negative infants but did not affect parenting satisfaction in mothers with less negative infants, consistent with a model of differential susceptibility. There was also a trend toward less weight gain in highly negative intervention group infants. In contrast, there was a main effect of the intervention on infant self-regulation at 1 year, such that the intervention group had higher observed self-regulation, across levels of infant negativity. Results support the importance of incorporating tests of moderation into evaluations of obesity interventions and also illustrate that individuals may be differentially susceptible to environmental effects on some outcomes but not others. PMID:23832637

  16. Mode-Stirred Method Implementation for HIRF Susceptibility Testing and Results Comparison with Anechoic Method

    NASA Technical Reports Server (NTRS)

    Nguyen, Truong X.; Ely, Jay J.; Koppen, Sandra V.

    2001-01-01

    This paper describes the implementation of mode-stirred method for susceptibility testing according to the current DO-160D standard. Test results on an Engine Data Processor using the implemented procedure and the comparisons with the standard anechoic test results are presented. The comparison experimentally shows that the susceptibility thresholds found in mode-stirred method are consistently higher than anechoic. This is consistent with the recent statistical analysis finding by NIST that the current calibration procedure overstates field strength by a fixed amount. Once the test results are adjusted for this value, the comparisons with the anechoic results are excellent. The results also show that test method has excellent chamber to chamber repeatability. Several areas for improvements to the current procedure are also identified and implemented.

  17. Dobutamine "stress" test and latent cardiac susceptibility to inhaled diesel exhaust in normal and hypertensive rats**

    EPA Science Inventory

    Background -Exercise "stress" testing is a screening tool used to determine the amount of stress for which the heart can compensate before developing abnormal rhythm or ischemia, particularly in susceptible people. Although this approach has been used to assess risk in humans exp...

  18. 21 CFR 866.1700 - Culture medium for antimicrobial susceptibility tests.

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ... 21 Food and Drugs 8 2012-04-01 2012-04-01 false Culture medium for antimicrobial susceptibility tests. 866.1700 Section 866.1700 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) MEDICAL DEVICES IMMUNOLOGY AND MICROBIOLOGY DEVICES Diagnostic Devices §...

  19. 21 CFR 866.1700 - Culture medium for antimicrobial susceptibility tests.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... 21 Food and Drugs 8 2010-04-01 2010-04-01 false Culture medium for antimicrobial susceptibility tests. 866.1700 Section 866.1700 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) MEDICAL DEVICES IMMUNOLOGY AND MICROBIOLOGY DEVICES Diagnostic Devices §...

  20. 21 CFR 866.1700 - Culture medium for antimicrobial susceptibility tests.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... 21 Food and Drugs 8 2011-04-01 2011-04-01 false Culture medium for antimicrobial susceptibility tests. 866.1700 Section 866.1700 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) MEDICAL DEVICES IMMUNOLOGY AND MICROBIOLOGY DEVICES Diagnostic Devices §...

  1. 21 CFR 866.1700 - Culture medium for antimicrobial susceptibility tests.

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ... 21 Food and Drugs 8 2013-04-01 2013-04-01 false Culture medium for antimicrobial susceptibility tests. 866.1700 Section 866.1700 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) MEDICAL DEVICES IMMUNOLOGY AND MICROBIOLOGY DEVICES Diagnostic Devices §...

  2. 21 CFR 866.1700 - Culture medium for antimicrobial susceptibility tests.

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ... 21 Food and Drugs 8 2014-04-01 2014-04-01 false Culture medium for antimicrobial susceptibility tests. 866.1700 Section 866.1700 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) MEDICAL DEVICES IMMUNOLOGY AND MICROBIOLOGY DEVICES Diagnostic Devices §...

  3. Antimicrobial susceptibility testing of Australian isolates of Brachyspira hyodysenteriae using a new broth dilution method.

    PubMed

    Karlsson, Märit; Oxberry, Sophy L; Hampson, David J

    2002-01-01

    The antimicrobial susceptibilities of 76 field isolates of Brachyspira hyodysenteriae from different states of Australia were tested in a newly developed broth dilution procedure. The antimicrobial agents used were tiamulin, valnemulin, tylosin, erythromycin, lincomycin and clindamycin. The results from the broth dilution susceptibility testing of 39 of the isolates were compared with results obtained for the same isolates using the agar dilution method. Amongst the isolates tested by broth dilution, 17 were from three farms and had been collected over a number of years. Their pulsed field gel electrophoresis pattern previously had been determined. The broth dilution technique was simple to use, less labor intensive than agar dilution, and gave clear end points. The results obtained using the two methods generally corresponded well, although in a few cases the MIC obtained by broth dilution were lower than those with agar dilution. For the 76 isolates tested by broth dilution, the MIC(90) (mg/l) was: tiamulin, 1; valnemulin, 0.5; tylosin>256; erythromycin>256; lincomycin, 64 and clindamycin, 16. Only minor differences in susceptibility patterns were found amongst isolates from different Australian states. Over all the isolates, and also amongst the isolates obtained from different years on the three farms, there was no trend for the susceptibility of the isolates to alter with time. PMID:11731165

  4. [Detection of cell death markers as a tool for bacterial antimicrobial susceptibility testing].

    PubMed

    Mlynárčik, P; Kolář, M

    2016-01-01

    Antimicrobial resistance among nosocomial pathogens has emerged as one of the most important health care problems in the new millennium. In this review, we present new methods for bacterial antimicrobial susceptibility testing, based on the detection of antibiotic-mediated cell death markers that could provide valuable alternatives to existing phenotypic approaches in the very near future. PMID:27467325

  5. Comparison of Three Reference Methods for Testing Susceptibility of Staphylococci to Trimethoprim-Sulfamethoxazole▿

    PubMed Central

    Griffith, R.; Creely, D.; Revell, P.; Dunne, W. Michael; Shortridge, D.

    2009-01-01

    Three reference MIC methods approved by the Clinical and Laboratory Standards Institute were compared by testing 567 staphylococci against trimethoprim-sulfamethoxazole. Category agreement ranged from 94.9% (broth macrodilution versus broth microdilution) to 98.6% (agar dilution versus broth microdilution). Twenty-seven strains resistant by broth macrodilution were susceptible by broth microdilution. PMID:19741069

  6. Multilaboratory Evaluation of In Vitro Antifungal Susceptibility Testing of Dermatophytes for ME1111.

    PubMed

    Ghannoum, M; Chaturvedi, V; Diekema, D; Ostrosky-Zeichner, L; Rennie, R; Walsh, T; Wengenack, N; Fothergill, A; Wiederhold, N

    2016-03-01

    ME1111 is a novel small molecule antifungal agent under development for the topical treatment of onychomycosis. Standardization of the susceptibility testing method for this candidate antifungal is needed. Toward this end, 8 independent laboratories determined the interlaboratory reproducibility of ME1111 susceptibility testing. In addition, we subsequently identified 2 strains as quality control (QC) isolates for the method. In the reproducibility study, 5 blinded clinical strains each of Trichophyton rubrum, Trichophyton mentagrophytes, and Epidermophyton floccosum were tested, while the QC study tested 6 blinded T. rubrum or T. mentagrophytes ATCC strains. Testing was performed in frozen microtiter panels according to the Clinical and Laboratory Standards Institute (CLSI) M38-A2 methodology. In the reproducibility study, 9 of 15 clinical strains showed interlaboratory agreement of >90% at the 80% inhibition endpoint, with a range of agreement of 76.2% to 100%. In the QC study, 4 of the 6 ATCC strains showed interlaboratory agreement of >90%. ME1111 demonstrated excellent interlaboratory agreement when tested against dermatophytes. Based on this data, the CLSI Subcommittee on Antifungal Susceptibility Tests approved the susceptibility testing of ME1111 against dermatophytes according to M38-A2 methodology, which stipulates RPMI 1640 as the test medium, an inoculum size of 1 to 3 × 10(3) CFU/ml, and an incubation time and temperature of 96 h at 35°C. The MIC endpoint should be 80% inhibition compared with the growth control. T. rubrum ATCC MYA-4438 and T. mentagrophytes ATCC 28185 were selected as QC isolates, with an acceptable range of 0.12 to 1 μg/ml for the two strains. PMID:26719434

  7. Antimicrobial Susceptibility Testing, Drug Resistance Mechanisms, and Therapy of Infections with Nontuberculous Mycobacteria

    PubMed Central

    Nash, Kevin A.; Wallace, Richard J.

    2012-01-01

    Summary: Within the past 10 years, treatment and diagnostic guidelines for nontuberculous mycobacteria have been recommended by the American Thoracic Society (ATS) and the Infectious Diseases Society of America (IDSA). Moreover, the Clinical and Laboratory Standards Institute (CLSI) has published and recently (in 2011) updated recommendations including suggested antimicrobial and susceptibility breakpoints. The CLSI has also recommended the broth microdilution method as the gold standard for laboratories performing antimicrobial susceptibility testing of nontuberculous mycobacteria. This article reviews the laboratory, diagnostic, and treatment guidelines together with established and probable drug resistance mechanisms of the nontuberculous mycobacteria. PMID:22763637

  8. Diagnosis of coccidioidomycosis by culture: safety considerations, traditional methods, and susceptibility testing.

    PubMed

    Sutton, Deanna A

    2007-09-01

    The recovery of Coccidioides spp. by culture and confirmation utilizing the AccuProbe nucleic acid hybridization method by GenProbe remain the definitive diagnostic method. Biosafety considerations from specimen collection through culture confirmation in the mycology laboratory are critical, as acquisition of coccidioidomycosis by laboratory workers is well documented. The designation of Coccidioides spp. as select agents of potential bioterrorism has mandated strict regulation of their transport and inventory. The genus appears generally susceptible, in vitro, although no defined breakpoints exist. Susceptibility testing may assist in documenting treatment failures. PMID:17363439

  9. Antimicrobial susceptibility testing, drug resistance mechanisms, and therapy of infections with nontuberculous mycobacteria.

    PubMed

    Brown-Elliott, Barbara A; Nash, Kevin A; Wallace, Richard J

    2012-07-01

    Within the past 10 years, treatment and diagnostic guidelines for nontuberculous mycobacteria have been recommended by the American Thoracic Society (ATS) and the Infectious Diseases Society of America (IDSA). Moreover, the Clinical and Laboratory Standards Institute (CLSI) has published and recently (in 2011) updated recommendations including suggested antimicrobial and susceptibility breakpoints. The CLSI has also recommended the broth microdilution method as the gold standard for laboratories performing antimicrobial susceptibility testing of nontuberculous mycobacteria. This article reviews the laboratory, diagnostic, and treatment guidelines together with established and probable drug resistance mechanisms of the nontuberculous mycobacteria. PMID:22763637

  10. Antimicrobial susceptibility testing of Neisseria gonorrhoeae and implications for epidemiology and therapy.

    PubMed Central

    Fekete, T

    1993-01-01

    Antimicrobial susceptibility testing (AST) of Neisseria gonorrhoeae has been under development since the early days of antimicrobial agents. However, it is rarely applied to clinical isolates today. The history of the various in vitro tests to determine the susceptibility of N. gonorrhoeae to antibiotics is rich with evidence that these results predict response to therapy for almost all agents tested. Further, AST is a useful and important aspect of strain characterization and disease epidemiology in conjunction with the more specific but laborious techniques of auxotyping, serotyping, and plasmid analysis. Current technology has overcome many of the objections to AST for N. gonorrhoeae with standardization of test media and the development of an accurate disk diffusion AST method that is suited to most clinical laboratories regardless of volume or level of technical expertise. Ironically, the very low level of resistance to the current primary treatment strategy in the United States, ceftriaxone or another potent cephalosporin, makes the use of AST somewhat superfluous. PMID:8457978

  11. Rapid bacterial antibiotic susceptibility test based on simple surface-enhanced Raman spectroscopic biomarkers

    PubMed Central

    Liu, Chia-Ying; Han, Yin-Yi; Shih, Po-Han; Lian, Wei-Nan; Wang, Huai-Hsien; Lin, Chi-Hung; Hsueh, Po-Ren; Wang, Juen-Kai; Wang, Yuh-Lin

    2016-01-01

    Rapid bacterial antibiotic susceptibility test (AST) and minimum inhibitory concentration (MIC) measurement are important to help reduce the widespread misuse of antibiotics and alleviate the growing drug-resistance problem. We discovered that, when a susceptible strain of Staphylococcus aureus or Escherichia coli is exposed to an antibiotic, the intensity of specific biomarkers in its surface-enhanced Raman scattering (SERS) spectra drops evidently in two hours. The discovery has been exploited for rapid AST and MIC determination of methicillin-susceptible S. aureus and wild-type E. coli as well as clinical isolates. The results obtained by this SERS-AST method were consistent with that by the standard incubation-based method, indicating its high potential to supplement or replace existing time-consuming methods and help mitigate the challenge of drug resistance in clinical microbiology. PMID:26997474

  12. SUSCEPTIBILITY TEST FOR FUNGI: CLINICAL AND LABORATORIAL CORRELATIONS IN MEDICAL MYCOLOGY

    PubMed Central

    ALASTRUEY-IZQUIERDO, Ana; MELHEM, Marcia S.C.; BONFIETTI, Lucas X.; RODRIGUEZ-TUDELA, Juan L.

    2015-01-01

    SUMMARY During recent decades, antifungal susceptibility testing has become standardized and nowadays has the same role of the antibacterial susceptibility testing in microbiology laboratories. American and European standards have been developed, as well as equivalent commercial systems which are more appropriate for clinical laboratories. The detection of resistant strains by means of these systems has allowed the study and understanding of the molecular basis and the mechanisms of resistance of fungal species to antifungal agents. In addition, many studies on the correlation of in vitro results with the outcome of patients have been performed, reaching the conclusion that infections caused by resistant strains have worse outcome than those caused by susceptible fungal isolates. These studies have allowed the development of interpretative breakpoints for Candida spp. and Aspergillus spp., the most frequent agents of fungal infections in the world. In summary, antifungal susceptibility tests have become essential tools to guide the treatment of fungal diseases, to know the local and global disease epidemiology, and to identify resistance to antifungals. PMID:26465371

  13. Is rapid antibacterial susceptibility testing medium reliable for routine laboratory practices?

    PubMed Central

    Yagmur, Gulhan; Ercal, Baris Derya; Mengeloglu, Zafer; Sariguzel, Fatma Mutlu; Berk, Elife; Saglam, Derya

    2015-01-01

    Objective: Early detection of antibiotic susceptibility profile of the isolates has critical importance in terms of immediate beginning of the appropriate treatment and increasing of treatment success, such as meningitis, bacteriemia and sepsis. In the present study, it was aimed to compare the antibiotic susceptibility results of Quicolor (Salubris Inc., Massachusetts, USA) and standard disk diffusion method. Methods: One hundred twenty three isolates were included in this study (80 Enterobacteriaceae, 15 Staphylococci and 28 nonfermentative Gram-negative bacteria). Antibiotic susceptibility in clinical isolates was evaluated using Mueller-Hinton (MH) agar and Quicolor (ES and NF) agar plates. Results: For Enterobacteriaceae, frequency of total concordance, major error, and minor error between the tests were found as 96.8%, 0.8%, and 2.4%, respectively. For Staphylococci, frequency of total concordance, major error, and minor error among the tests were found as 95.7%, 3.5%, and 0.8%, respectively. For non fermentative bacteria, frequency of total concordance, major error, and minor error among the tests were found as 83.9%, 9.6%, and 6.4%, respectively. Conclusions: Quicolor media provided reliable susceptibility results in enteric bacteria and Staphylococci. However, further studies including higher number of nonfermentative bacteria are required to determine whether the chromogenic media are appropriate for this group of bacteria. PMID:26101489

  14. Frequency of Susceptibility Testing for Patients with Persistent Methicillin-Resistant Staphylococcus aureus Bacteremia

    PubMed Central

    Giltner, Carmen L.; Kelesidis, Theodoros; Hindler, Janet A.; Bobenchik, April M.

    2014-01-01

    Currently, no standards exist for determining the optimal frequency of repeat antimicrobial susceptibility testing (AST) when an organism is recurrently isolated from the same specimen source. Although testing every 2 to 5 days is thought sufficient, we present three cases of methicillin-resistant Staphylococcus aureus (MRSA) bacteremia where current laboratory protocol for repeating AST every 5 days was inadequate to identify resistant organisms. PMID:24153125

  15. A Reference Broth Microdilution Method for Dalbavancin In Vitro Susceptibility Testing of Bacteria that Grow Aerobically.

    PubMed

    Koeth, Laura M; DiFranco-Fisher, Jeanna M; McCurdy, Sandra

    2015-01-01

    Antimicrobial susceptibility testing (AST) is performed to assess the in vitro activity of antimicrobial agents against various bacteria. The AST results, which are expressed as minimum inhibitory concentrations (MICs) are used in research for antimicrobial development and monitoring of resistance development and in the clinical setting for antimicrobial therapy guidance. Dalbavancin is a semi-synthetic lipoglycopeptide antimicrobial agent that was approved in May 2014 by the Food and Drug Administration (FDA) for the treatment of acute bacterial skin and skin structure infections caused by Gram-positive organisms. The advantage of dalbavancin over current anti-staphylococcal therapies is its long half-life, which allows for once-weekly dosing. Dalbavancin has activity against Staphylococcus aureus (including both methicillin-susceptible S. aureus [MSSA] and methicillin-resistant S. aureus [MRSA]), coagulase-negative staphylococci, Streptococcus pneumoniae, Streptococcus anginosus group, β-hemolytic streptococci and vancomycin susceptible enterococci. Similar to other recent lipoglycopeptide agents, optimization of CLSI and ISO broth susceptibility test methods includes the use of dimethyl sulfoxide (DMSO) as a solvent when preparing stock solutions and polysorbate 80 (P80) to alleviate adherence of the agent to plastic. Prior to the clinical studies and during the initial development of dalbavancin, susceptibility studies were not performed with the use of P-80 and MIC results tended to be 2-4 fold higher and similarly higher MIC results were obtained with the agar dilution susceptibility method. Dalbavancin was first included in CLSI broth microdilution methodology tables in 2005 and amended in 2006 to clarify use of DMSO and P-80. The broth microdilution (BMD) procedure shown here is specific to dalbavancin and is in accordance with the CLSI and ISO methods, with step-by-step detail and focus on the critical steps added for clarity. PMID:26381422

  16. Comparison of E-test with agar dilution methods in testing susceptibility of N. gonorrhoeae to azithromycin.

    PubMed

    Yasin, R M; Suan, K A; Meng, C Y

    1997-05-01

    A single dose of a new antibiotic, azithromycin, has been shown to be effective in the treatment of uncomplicated Neisseria gonorrhoeae. A clinical study was conducted to assess the in vitro susceptibility of N gonorrhoeae to azithromycin and compare the reliability of results obtained using the new E-test methodology for determination of the minimum inhibitory concentration (MIC) of antibiotic with those obtained through the standard agar dilution method. 135 clinical isolates of N gonorrhoeae were obtained from patients attending hospital-based sexually transmitted disease clinics in five geographic locations in Malaysia. 76 of the isolates were penicillinase-producing N gonorrhoeae and 69 were high-level tetracycline-resistant N gonorrhoeae. All isolates were susceptible to azithromycin based on the susceptible MIC breakpoint of 2.0 mcg/ml. The MICs ranged from 0.0078-0.25 mcg/ml by agar dilution method and from 0.016-0.50 mcg/ml by E-test. Agreement between these two methods was 97.8%. The single-dose regime and good antigonococcal and antichlamydial activity of azithromycin make this antibiotic a suitable treatment choice. Moreover, the findings of this study suggest that the simpler, faster E-test is as reliable as the agar dilution method. Given the tendency of the antimicrobial susceptibility pattern of N gonorrhoeae to change rapidly, it is important to monitor MICs to detect the emergence of resistance. PMID:9153733

  17. A microfluidic platform for rapid, stress-induced antibiotic susceptibility testing of Staphylococcus aureus

    PubMed Central

    Kalashnikov, Maxim; Lee, Jean C.; Campbell, Jennifer; Sharon, Andre; Sauer-Budge, Alexis F.

    2012-01-01

    The emergence and spread of bacterial resistance to ever increasing classes of antibiotics intensifies the need for fast phenotype-based clinical tests for determining antibiotic susceptibility. Standard susceptibility testing relies on the passive observation of bacterial growth inhibition in the presence of antibiotics. In this paper, we present a novel microfluidic platform for antibiotic susceptibility testing basedon stress-activation of biosynthetic pathways that are the primary targets of antibiotics. We chose Staphylococcus aureus as a model system due to its clinical importance, and we selected bacterial cell wall biosynthesis as the primary target of both stress and antibiotic. Enzymatic and mechanical stresses were used to damage the bacterial cell wall, and a β-lactam antibiotic interfered with the repair process, resulting in rapid cell death of strains that harbor no resistance mechanism. In contrast, resistant bacteria remained viable under the assay conditions. Bacteria, covalently-bound to the bottom of the microfluidic channel, were subjected to mechanical shear stress created by flowing culture media through the microfluidic channel and to enzymatic stress with sub-inhibitory concentrations of the bactericidal agent lysostaphin. Bacterial cell death was monitored via fluorescence using the Sytox Green dead cell stain, and rates of killing were measured for the bacterial samples in the presence and absence of oxacillin. Using model susceptible (Sanger 476) and resistant (MW2) S. aureus strains, a metric was established to separate susceptible and resistant staphylococci based on normalized fluorescence values after 60 minutes of exposure to stress and antibiotic. Because this groundbreaking approach is not based on standard methodology, it circumvents the need for minimum inhibitory concentration (MIC) measurements and long wait times. We demonstrate the successful development of a rapid microfluidic-based and stress-activated antibiotic

  18. Dynamic Biomechanical Examination of the Lumbar Spine with Implanted Total Disc Replacement (TDR) Utilizing a Pendulum Testing System

    PubMed Central

    Daniels, Alan H; Paller, David J; Koruprolu, Sarath; McDonnell, Matthew; Palumbo, Mark A; Crisco, Joseph J

    2013-01-01

    Study Design Biomechanical cadaver investigation Objective To examine dynamic bending stiffness and energy absorption of the lumbar spine with and without implanted Total Disc Replacement (TDR) under simulated physiologic motion. Summary of background data The pendulum testing system is capable of applying physiologic compressive loads without constraining motion of functional spinal units (FSUs). The number of cycles to equilibrium observed under pendulum testing is a measure of the energy absorbed by the FSU. Methods Five unembalmed, frozen human lumbar FSUs were tested on the pendulum system with axial compressive loads of 181N, 282N, 385N, and 488N before and after Synthes ProDisc-L TDR implantation. Testing in flexion, extension, and lateral bending began by rotating the pendulum to 5° resulting in unconstrained oscillatory motion. The number of rotations to equilibrium was recorded and bending stiffness (N-m/°) was calculated and compared for each testing mode. Results In flexion/extension, the TDR constructs reached equilibrium with significantly (p<0.05) fewer cycles than the intact FSU with compressive loads of 282N, 385N and 488N. Mean dynamic bending stiffness in flexion, extension, and lateral bending increased significantly with increasing load for both the intact FSU and TDR constructs (p<0.001). In flexion, with increasing compressive loading from 181N to 488N, the bending stiffness of the intact FSUs increased from 4.0N-m/° to 5.5N-m/°, compared to 2.1N-m/° to 3.6N-m/° after TDR implantation. At each compressive load, the intact FSU was significantly more stiff than the TDR (p<0.05). Conclusion Lumbar FSUs with implanted TDR were found to be less stiff, but also absorbed more energy during cyclic loading with an unconstrained pendulum system. Although the effects on clinical performance of motion preserving devices are not fully known, these results provide further insight into the biomechanical behavior of this device under approximated

  19. Patients' attitudes about autonomy and confidentiality in genetic testing for breast-ovarian cancer susceptibility.

    PubMed

    Benkendorf, J L; Reutenauer, J E; Hughes, C A; Eads, N; Willison, J; Powers, M; Lerman, C

    1997-12-19

    The identification of BRCA1 and BRCA2, two breast-ovarian cancer susceptibility genes, has brought many ethical and social issues to the forefront. This paper presents the results of a survey assessing the attitudes of 238 unaffected first-degree relatives of women with breast or ovarian cancer regarding the ethical issues of autonomy and confidentiality as they relate to BRCA1/2 testing. Baseline knowledge about BRCA1/2 and ethnic and psychosocial characteristics of our study population were examined to determine their association with women's attitudes. The majority of women (86-87%) felt that health care providers should not disclose the results of genetic tests for breast-ovarian cancer susceptibility to insurance companies or employers without written consent; however, only 56-57% felt that written consent should be required for a spouse or immediate family to receive this information. Ninety-eight percent of the women surveyed agreed that genetic testing for breast-ovarian cancer risk should be voluntary. Likewise, most women (95%) agreed that a person should be able to have genetic testing against a doctor's recommendation and 88% of the women surveyed agreed that parents should be able to consent to genetic susceptibility testing on behalf of their minor children. African American women were less concerned than Caucasian women about the protection of confidentiality in families, they were more likely to agree that an individual should still have access to testing when their physicians recommended against it, and they were more supportive of parents' rights to consent to genetic predisposition testing on behalf of their minor children. Women with coping styles characterized by higher optimism were more likely to favor access to genetic testing when a physician recommended against it, and to support parents' rights to consent to testing of their minor children. Therefore, the setting and manner in which genetic counseling and testing are delivered must be

  20. Liofilchem® Chromatic VRE and vancomycin MIC Test Strip detected glycopeptide resistance in a vanB neonatal Enterococcus faecium isolate showing alternate vancomycin susceptibility and resistance with bioMérieux Vitek2

    PubMed Central

    Savini, Vincenzo; Marrollo, Roberta; Coclite, Eleonora; Fusilli, Paola; D’Incecco, Carmine; Fazii, Paolo; Gherardi, Giovanni

    2014-01-01

    A 1-month old neonate urine sample yielded vanB Enterococcus faecium; nevertheless, the isolate alternatively showed susceptibility and resistance to vancomycin with bioMérieux Vitek2 (cards AST592, AST632, AST586), while glycopeptide resistance was detected by Liofilchem® vancomycin MIC Test Strip and disc along with the Chromatic VRE chromogenic medium. This communication emphasizes that, as vanB gene may be heterogeneously expressed within a given Enterococcus population, glycopeptide resistance may be missed when using automated systems for antibiotic susceptibility testing. We suggest therefore that vancomycin in vitro activity be studied on all clinical isolates through agar methods, including use of chromogenic media. PMID:25337280

  1. Liofilchem(®) Chromatic VRE and vancomycin MIC Test Strip detected glycopeptide resistance in a vanB neonatal Enterococcus faecium isolate showing alternate vancomycin susceptibility and resistance with bioMérieux Vitek2.

    PubMed

    Savini, Vincenzo; Marrollo, Roberta; Coclite, Eleonora; Fusilli, Paola; D'Incecco, Carmine; Fazii, Paolo; Gherardi, Giovanni

    2014-01-01

    A 1-month old neonate urine sample yielded vanB Enterococcus faecium; nevertheless, the isolate alternatively showed susceptibility and resistance to vancomycin with bioMérieux Vitek2 (cards AST592, AST632, AST586), while glycopeptide resistance was detected by Liofilchem(®) vancomycin MIC Test Strip and disc along with the Chromatic VRE chromogenic medium. This communication emphasizes that, as vanB gene may be heterogeneously expressed within a given Enterococcus population, glycopeptide resistance may be missed when using automated systems for antibiotic susceptibility testing. We suggest therefore that vancomycin in vitro activity be studied on all clinical isolates through agar methods, including use of chromogenic media. PMID:25337280

  2. Multicenter evaluation of the MB/BACT system for susceptibility testing of Mycobacterium tuberculosis.

    PubMed

    Bemer, Pascale; Bodmer, Thomas; Munzinger, Juerg; Perrin, Monique; Vincent, Véronique; Drugeon, Henri

    2004-03-01

    The reliability of the MB/BACT system for susceptibility testing of Mycobacterium tuberculosis to pyrazinamide, rifampin, isoniazid, streptomycin, and ethambutol was compared to the BACTEC 460TB system. The proportion method was used to resolve discrepant results by an independent arbiter. Two interpretative methods were used, with an undiluted control (direct control) and a diluted control (10(-1) control). As no significant difference was observed between the two controls, the method with the direct control was adopted as the most accurate one. One hundred sixty-six strains were tested, with an overall agreement of 98.3%. After resolution of the 18 discrepant results by the proportion method, the sensitivity and specificity of the MB/BACT system were 100% for rifampin, isoniazid, and pyrazinamide. For ethambutol, sensitivity was 92.3% at the critical concentration and 33% at the high concentration, and specificity was 100% at both concentrations. For streptomycin, sensitivity was 100% at the critical concentration and 80% at the high concentration, and specificity was 98.6% at the critical concentration and 100% at the high concentration. The rifampin, isoniazid, streptomycin, and ethambutol susceptibility test results were obtained in 6.6 days with the MB/BACT versus 5 days with the BACTEC 460TB. The pyrazinamide susceptibility test results were obtained in 7.8 days with the MB/BACT, versus 6.7 days with the BACTEC 460TB. These data demonstrate that the fully automated MB/BACT system is a very reliable method for rapid susceptibility testing of M. tuberculosis against rifampin, isoniazid, and pyrazinamide. Sensitivity results have to be improved for ethambutol and streptomycin, especially at the high concentration. PMID:15004049

  3. Multicenter Evaluation of the MB/BACT System for Susceptibility Testing of Mycobacterium tuberculosis

    PubMed Central

    Bemer, Pascale; Bodmer, Thomas; Munzinger, Juerg; Perrin, Monique; Vincent, Véronique; Drugeon, Henri

    2004-01-01

    The reliability of the MB/BACT system for susceptibility testing of Mycobacterium tuberculosis to pyrazinamide, rifampin, isoniazid, streptomycin, and ethambutol was compared to the BACTEC 460TB system. The proportion method was used to resolve discrepant results by an independent arbiter. Two interpretative methods were used, with an undiluted control (direct control) and a diluted control (10−1 control). As no significant difference was observed between the two controls, the method with the direct control was adopted as the most accurate one. One hundred sixty-six strains were tested, with an overall agreement of 98.3%. After resolution of the 18 discrepant results by the proportion method, the sensitivity and specificity of the MB/BACT system were 100% for rifampin, isoniazid, and pyrazinamide. For ethambutol, sensitivity was 92.3% at the critical concentration and 33% at the high concentration, and specificity was 100% at both concentrations. For streptomycin, sensitivity was 100% at the critical concentration and 80% at the high concentration, and specificity was 98.6% at the critical concentration and 100% at the high concentration. The rifampin, isoniazid, streptomycin, and ethambutol susceptibility test results were obtained in 6.6 days with the MB/BACT versus 5 days with the BACTEC 460TB. The pyrazinamide susceptibility test results were obtained in 7.8 days with the MB/BACT, versus 6.7 days with the BACTEC 460TB. These data demonstrate that the fully automated MB/BACT system is a very reliable method for rapid susceptibility testing of M. tuberculosis against rifampin, isoniazid, and pyrazinamide. Sensitivity results have to be improved for ethambutol and streptomycin, especially at the high concentration. PMID:15004049

  4. American Society of Clinical Oncology Policy Statement Update: Genetic and Genomic Testing for Cancer Susceptibility.

    PubMed

    Robson, Mark E; Bradbury, Angela R; Arun, Banu; Domchek, Susan M; Ford, James M; Hampel, Heather L; Lipkin, Stephen M; Syngal, Sapna; Wollins, Dana S; Lindor, Noralane M

    2015-11-01

    The American Society of Clinical Oncology (ASCO) has long affirmed that the recognition and management of individuals with an inherited susceptibility to cancer are core elements of oncology care. ASCO released its first statement on genetic testing in 1996 and updated that statement in 2003 and 2010 in response to developments in the field. In 2014, the Cancer Prevention and Ethics Committees of ASCO commissioned another update to reflect the impact of advances in this area on oncology practice. In particular, there was an interest in addressing the opportunities and challenges arising from the application of massively parallel sequencing-also known as next-generation sequencing-to cancer susceptibility testing. This technology introduces a new level of complexity into the practice of cancer risk assessment and management, requiring renewed effort on the part of ASCO to ensure that those providing care to patients with cancer receive the necessary education to use this new technology in the most effective, beneficial manner. The purpose of this statement is to explore the challenges of new and emerging technologies in cancer genetics and provide recommendations to ensure their optimal deployment in oncology practice. Specifically, the statement makes recommendations in the following areas: germline implications of somatic mutation profiling, multigene panel testing for cancer susceptibility, quality assurance in genetic testing, education of oncology professionals, and access to cancer genetic services. PMID:26324357

  5. Electromagnetic Compatibility Testing for Conducted Susceptibility Along Interconnecting Signal Lines. Final report

    SciTech Connect

    Ewing, P. D.; Wood, R. T.; Korsah, K.; Shourbaji, A. A.; Wilson, T. L.; Beets, B. M.

    2002-07-31

    This document presents recommendations and the associated technical basis for addressing the effects of conducted electromagnetic interference (EMI) and radio-frequency interference (RFI) along interconnecting signal lines in safety-related instrumentation and control (I&C) systems. Oak Ridge National Laboratory has been engaged in assisting the U.S. Nuclear Regulatory Commission Office of Nuclear Regulatory Research in developing the technical basis for regulatory guidance on EMIIRFI immunity and power surge withstand capability (SWC). Previous research efforts have provided recommendations on (1) electromagnetic compatibility design and installation practices, (2) the endorsement of EMI/RFI and SWC test criteria and test methods, (3) the determination of ambient electromagnetic conditions at nuclear power plants, and (4) the development of recommended electromagnetic operating envelopes applicable to locations where safety-related I&C systems will be installed. The current research focuses on the susceptibility of l&C systems to conducted EMIIRFI along interconnecting signal lines. Coverage of signal line susceptibility was identified as an open issue in previous research on establishing the technical basis for EMIIRFI and SWC in safety-related I&C systems. Research results provided in this report will be used to establish the technical basis for endorsing U.S. Department of Defense and European Committee for Electrotechnical Standardization test criteria and test methods that address signal-line susceptibility. In addition, recommendations on operating envelopes are presented based on available technical information.

  6. A rapid antimicrobial susceptibility test based on single-cell morphological analysis.

    PubMed

    Choi, Jungil; Yoo, Jungheon; Lee, Mincheol; Kim, Eun-Geun; Lee, Ji Soo; Lee, Seungok; Joo, Seik; Song, Sang Hoon; Kim, Eui-Chong; Lee, Jung Chan; Kim, Hee Chan; Jung, Yong-Gyun; Kwon, Sunghoon

    2014-12-17

    A rapid antibiotic susceptibility test (AST) is desperately needed in clinical settings for fast and appropriate antibiotic administration. Traditional ASTs, which rely on cell culture, are not suitable for urgent cases of bacterial infection and antibiotic resistance owing to their relatively long test times. We describe a novel AST called single-cell morphological analysis (SCMA) that can determine antimicrobial susceptibility by automatically analyzing and categorizing morphological changes in single bacterial cells under various antimicrobial conditions. The SCMA was tested with four Clinical and Laboratory Standards Institute standard bacterial strains and 189 clinical samples, including extended-spectrum β-lactamase-positive Escherichia coli and Klebsiella pneumoniae, imipenem-resistant Pseudomonas aeruginosa, methicillin-resistant Staphylococcus aureus, and vancomycin-resistant Enterococci from hospitals. The results were compared with the gold standard broth microdilution test. The SCMA results were obtained in less than 4 hours, with 91.5% categorical agreement and 6.51% minor, 2.56% major, and 1.49% very major discrepancies. Thus, SCMA provides rapid and accurate antimicrobial susceptibility data that satisfy the recommended performance of the U.S. Food and Drug Administration. PMID:25520395

  7. Genotyping and In Vitro Antifungal Susceptibility Testing of Fusarium Isolates from Onychomycosis in India.

    PubMed

    Gupta, Chhavi; Jongman, Marit; Das, Shukla; Snehaa, K; Bhattacharya, S N; Seyedmousavi, S; van Diepeningen, Anne D

    2016-08-01

    Onychomycosis refers to fungal infection of the nail and is commonly caused by dermatophytes, while yeasts and non-dermatophytic molds (NDM) are increasingly recognized as pathogens in nail infections. The present study was done to delineate molecular epidemiology of Fusarium onychomycosis in India. Five hundred nail samples of Indian patients clinically suspected of onychomycosis were subjected to direct microscopy and fungal culture. Representative Fusarium isolates were further identified to species level by multi-locus sequencing for internal transcribed spacer, translation elongation factor 1 alpha (tef1-α) and RNA polymerase II subunit (rpb2) regions (primer pairs: ITS1/ITS4, EF1/EF2, 5f2/7cr, respectively). These representative strains were also tested for in vitro antifungal susceptibility by the broth microdilution method. Members of the genus Fusarium proved to be the most common NDM responsible for onychomycosis. The Fusarium spp. responsible for onychomycosis belonged to the Fusarium solani species complex (F. keratoplasticum and F. falciforme) and Fusarium fujikuroi species complex (F. proliferatum, F. acutatum and F. sacchari). Antifungal susceptibility results indicated that amphotericin B was the most effective antifungal across all isolates (MIC ranging 0.5-2 mg/L), followed by voriconazole (MIC ranging 1-8 µg/ml). However, a large variation was shown in susceptibility to posaconazole (MIC ranging 0.5 to >16 µg/ml). To conclude, we identified different Fusarium spp. responsible for onychomycosis in India with variation within species in susceptibility to antifungal agents, showing that fusariosis requires correct and prompt diagnosis as well as antifungal susceptibility testing. PMID:27138574

  8. Development of an Accelerated Test Method for the Determination of Susceptibility to Atmospheric Corrosion

    NASA Technical Reports Server (NTRS)

    Ambrose, John R.

    1991-01-01

    The theoretical rationale is presented for use of a repetitive cyclic current reversal voltammetric technique for characterization of localized corrosion processes, including atmospheric corrosion. Applicability of this proposed experimental protocol is applied to characterization of susceptibility to crevice and pitting corrosion, atmospheric corrosion and stress corrosion cracking. Criteria upon which relative susceptibility is based were determined and tested using two iron based alloys commonly in use at NASA-Kennedy; A36 (a low carbon steel) and 4130 (a low alloy steel). Practicality of the procedure was demonstrated by measuring changes in anodic polarization behavior during high frequency current reversal cycles of 25 cycles per second with 1 mA/sq cm current density amplitude in solutions containing Cl anions. The results demonstrated that, due to excessive polarization which affects conductivity of barrier corrosion product layers, A36 was less resistant to atmospheric corrosion than its 4130 counterpart; behavior which was also demonstrated during exposure tests.

  9. Growth inhibition of Mycobacterium tuberculosis by polyoxyethylene stearate present in the BACTEC pyrazinamide susceptibility test.

    PubMed

    Miller, M A; Thibert, L; Desjardins, F; Siddiqi, S H; Dascal, A

    1996-01-01

    We have previously found that approximately 3.5% of 428 clinical isolates of Mycobacterium tuberculosis yield uninterpretable results in the BACTEC pyrazinamide (PZA) susceptibility test system, because of inadequate growth. We tested the hypothesis that polyoxyethylene stearate (POES), the ingredient of the reconstituting fluid for the test, was the cause of this growth inhibition. A total of 15 isolates known for their previously uninterpretable results and 100 randomly chosen clinical isolates were tested in parallel both with and without POES. Repeat testing of the isolates with previously uninterpretable results yielded results in the presence of POES in only seven (47%). In the absence of POES, all gave interpretable results but one such result showed false resistance. For the other 100 clinical isolates, interpretable results were obtained with and without POES, but growth was enhanced in the absence of POES, especially in the PZA-susceptible strains. This was evidenced by a decreased time to attain a growth index of 200 in the control vial (4.9 days without POES versus 5.8 days with POES; P < 0.001) and a higher mean growth index ratio on the day of interpretation of the test (7.4% without POES versus 2.2% with POES; P < 0.001). However, the enhanced growth without POES led to 20 susceptible strains being misinterpreted as either resistant or borderline. We suggest that isolates of M. tuberculosis which yield uninterpretable results in the BACTEC PZA test system should be retested both with and without POES. If interpretable results indicating PZA resistance are obtained only in the absence of POES, the result should be confirmed by a pyrazinamidase assay or by the conventional proportion method. Routine omission of POES from the BACTEC test for all clinical strains is discouraged because of the unacceptably high false-resistance rates. PMID:8748279

  10. Correlation of different phenotypic drug susceptibility testing methods for four fluoroquinolones in Mycobacterium tuberculosis

    PubMed Central

    Coeck, Nele; de Jong, Bouke C.; Diels, Maren; de Rijk, Pim; Ardizzoni, Elisa; Van Deun, Armand; Rigouts, Leen

    2016-01-01

    Background Molecular resistance testing fails to explain all fluoroquinolone resistance, with a continued need for a suitable rapid phenotypic drug susceptibility testing method. Objective To evaluate the optimal method for phenotypic fluoroquinolone susceptibility testing. Methods Using Löwenstein–Jensen medium, Middlebrook 7H11 agar, BACTEC-MGIT 960 and the resazurin microtitre plate assay, we determined susceptibility to fluoroquinolones in Mycobacterium tuberculosis and investigated cross-resistance between ofloxacin, levofloxacin, moxifloxacin and gatifloxacin. We compared MICs of all four fluoroquinolones for 91 strains on Löwenstein–Jensen (as the gold standard) with their MICs in resazurin plates, and with ofloxacin susceptibility at a single concentration in MGIT and on 7H11 agar, in addition to sequencing of the gyrAB genes. Results and conclusions Applying a cut-off of 2 mg/L ofloxacin, 1 mg/L levofloxacin and 0.5 mg/L moxifloxacin and gatifloxacin in all methods, some discordance between solid medium and MGIT methods was observed, yet this tended to be explained by MICs around the cut-off. The high discordance between Löwenstein–Jensen (LJ) and resazurin plates suggests that the currently applied cut-offs for all fluoroquinolones in the resazurin method should decrease and minor changes in colour (from blue to purple) be considered as meaningful. High-level resistance in all assays to all drugs correlated well with the presence of gyrA mutations, in support of recent findings that fluoroquinolone resistance should be tested at different concentrations, as patients with lower levels of resistance may continue to benefit from high-dose fluoroquinolone-based therapy. PMID:26851609

  11. Single event upset susceptibility testing of the Xilinx Virtex II FPGA

    NASA Technical Reports Server (NTRS)

    Yui, C.; Swift, G.; Carmichael, C.

    2002-01-01

    Heavy ion testing of the Xilinx Virtex IZ was conducted on the configuration, block RAM and user flip flop cells to determine their single event upset susceptibility using LETs of 1.2 to 60 MeVcm^2/mg. A software program specifically designed to count errors in the FPGA is used to reveal L1/e values and single-event-functional interrupt failures.

  12. Antifungal Susceptibility Testing of Fluconazole by Flow Cytometry Correlates with Clinical Outcome

    PubMed Central

    Wenisch, Christoph; Moore, Caroline B.; Krause, Robert; Presterl, Elisabeth; Pichna, Peter; Denning, David W.

    2001-01-01

    Susceptibility testing of fungi by flow cytometry (also called fluorescence-activated cell sorting [FACS]) using vital staining with FUN-1 showed a good correlation with the standard M27-A procedure for assessing MICs. In this study we determined MICs for blood culture isolates from patients with candidemia by NCCLS M27-A and FACS methods and correlated the clinical outcome of these patients with in vitro antifungal resistance test results. A total of 24 patients with candidemia for whom one or more blood cultures were positive for a Candida sp. were included. Susceptibility testing was performed by NCCLS M27-A and FACS methods. The correlation of MICs (NCCLS M27-A and FACS) and clinical outcome was calculated. In 83% of the cases, the MICs of fluconazole determined by FACS were within 1 dilution of the MICs determined by the NCCLS M27-A method. For proposed susceptibility breakpoints, there was 100% agreement between the M27-A and FACS methods. In the FACS assay, a fluconazole MIC of <1 μg/ml was associated with cure (P < 0.001) whereas an MIC of ≥1 μg/ml was associated with death (P < 0.001). The M27-A-derived fluconazole MICs did not correlate with outcome (P = 1 and P = 0.133). PMID:11427554

  13. Experimental Study of the Rolling-Sliding Contact Conditions in a PA66/STEEL Gear Using Twin-Disc Test Rig: Friction and Wear Analysis

    NASA Astrophysics Data System (ADS)

    Mbarek, Meftah; Rhaiem, Sadok; Kharrat, Mohamed; Dammak, Maher

    2015-09-01

    This study investigates the effects of sliding ratio on the tribological response of the contact between the teeth of a metal/polymer gear in the regions close to the pitch point. For this purpose, a new twin-disc test rig was developed on the basis of two discs of different diameters rotating one above the other at the same angular speed. Two different materials were used: non-alloyed structural steel (C45) and polyamide (PA66). The effect of the slip ratio (4%, 12%, 20% and 28%) was studied at a constant pressure of 34 MPa and a constant angular speed of 300 rpm. In addition, the contact conditions were controlled with measurements of the two discs surface temperatures. The results indicate that the wear and the friction are closely related to the contact temperature generated by the sliding phenomenon. At low slip ratio (4% and 12%), the coefficient of friction and the temperature are characterized by a quasi-linear increase with time, and the wear increases slowly. At higher slip ratio (20% and 28%), the coefficient of friction and the temperature presents a steady state, and the wear increases dramatically. During the test, a film of transferred PA66 is formed on the steel surface causing the development of adhesive interactions between the contacting discs which increase the friction coefficient and the contact temperature. The high thermal conductivity of steel as compared to that of the polymer can reduce enormously the contact temperature generated by the sliding process.

  14. Current perspectives on tigecycline resistance in Enterobacteriaceae: susceptibility testing issues and mechanisms of resistance.

    PubMed

    Pournaras, Spyros; Koumaki, Vasiliki; Spanakis, Nicholas; Gennimata, Vasiliki; Tsakris, Athanassios

    2016-07-01

    During the past decades, rates of multidrug-resistant (MDR) and carbapenem-resistant (CR) Enterobacteriaceae clinical isolates, mainly Klebsiella spp., Escherichia coli, Enterobacter spp., Proteus spp. and Serratia marcescens, have increased, considerably restricting effective antimicrobial treatments. Tigecycline, the first member of the glycylcyclines, has been approved by the US Food and Drug Administration (FDA) for the treatment of complicated skin and soft-tissue, complicated intra-abdominal and community-acquired bacterial respiratory infections and is increasingly administered against MDR Enterobacteriaceae. Although resistance has gradually appeared, tigecycline still remains relatively active among Enterobacteriaceae, with resistance rates largely <10% in most wide-scale surveillance studies. Tigecycline resistance has been reported in some studies to be elevated among extended-spectrum β-lactamase (ESBL)-producing, MDR, extensively drug-resistant and CR isolates. Broth microdilution (BMD) is the reference method for tigecycline susceptibility testing, but disagreements have been reported between the methods applied for routine tigecycline susceptibility testing. Therefore, confirmation of daily tigecycline susceptibility testing with BMD appears important in order to avoid misclassification of isolates. Various mechanisms have been reported to confer tigecycline resistance, with RND-type transporters, mainly the AcrAB efflux pump, playing an important role. Other pumps and various control pathways are also implicated in tigecycline resistance. Overall, tigecycline is a potent therapeutic option for enterobacterial infections. Accurate detection of tigecycline susceptibility status and surveillance of resistant organisms in the hospital environment is necessary in order to optimise its use and to preserve tigecycline in our therapeutic arsenal. PMID:27256586

  15. Multicenter Evaluation of a Nonweekend Reading Schedule for Radiometric Pyrazinamide Susceptibility Testing of Mycobacterium tuberculosis

    PubMed Central

    Madison, B.; Gross, W.; George, I.; Sloutsky, A.; Washabaugh, G.; Robinson-Dunn, B.; Lipman, H.; Metchock, B.; Mazurek, G.; Ridderhof, J.

    2002-01-01

    Pyrazinamide (PZA) is an integral component of the short-course chemotherapy regimen for tuberculosis. The BACTEC 460TB PZA susceptibility test for Mycobacterium tuberculosis with a daily (D) reading schedule has been available for more than 10 years, but weekend laboratory staffing is necessary. A nonweekend (NW) reading schedule has not been validated in a multicenter study. This prospective multicenter study compares the interlaboratory reproducibility of PZA susceptibility results by following both the D and NW schedules. A total of 181 cultures were shared among four laboratories. Isolates were selected based on resistance or borderline resistance to at least one streptomycin-isoniazid-rifampin-ethambutol drug or PZA. One laboratory used a D reading schedule, and three laboratories used a NW schedule. Both reading schedules are based on the standard BACTEC 460TB PZA protocol. With the NW schedule, the growth index (GI) is not available for test interpretation on Saturday, Sunday, and Monday. Of the 181 shared cultures, 154 were found to be susceptible by all laboratories, 19 were found to be resistant, and 8 had discordant results. The overall pairwise interlaboratory agreement was 97.7%. The discrepancies were not associated with the type of reading schedule used. However, the median control GI was significantly higher for the NW schedule (321) than for the D schedule (259) (P < 0.0001) although results were available on average in about 7 days from setup for both schedules. These results show that the NW schedule is a suitable alternative for laboratories that do not read and interpret PZA susceptibility tests on weekends. PMID:12354876

  16. Comparing Etest and Broth Microdilution for Antifungal Susceptibility Testing of the Most-Relevant Pathogenic Molds

    PubMed Central

    Lamoth, Frédéric

    2015-01-01

    Invasive mold infections are life-threatening diseases for which appropriate antifungal therapy is crucial. Their epidemiology is evolving, with the emergence of triazole-resistant Aspergillus spp. and multidrug-resistant non-Aspergillus molds. Despite the lack of interpretive criteria, antifungal susceptibility testing of molds may be useful in guiding antifungal therapy. The standard broth microdilution method (BMD) is demanding and requires expertise. We assessed the performance of a commercialized gradient diffusion method (Etest method) as an alternative to BMD. The MICs or minimal effective concentrations (MECs) of amphotericin B, voriconazole, posaconazole, caspofungin, and micafungin were assessed for 290 clinical isolates of the most representative pathogenic molds (154 Aspergillus and 136 non-Aspergillus isolates) with the BMD and Etest methods. Essential agreements (EAs) within ±2 dilutions of ≥90% between the two methods were considered acceptable. EAs for amphotericin B and voriconazole were >90% for most potentially susceptible species. For posaconazole, the correlation was acceptable for Mucoromycotina but Etest MIC values were consistently lower for Aspergillus spp. (EAs of <90%). Excellent EAs were found for echinocandins with highly susceptible (MECs of <0.015 μg/ml) or intrinsically resistant (MECs of >16 μg/ml) strains. However, MEC determinations lacked consistency between methods for strains exhibiting mid-range MECs for echinocandins. We concluded that the Etest method is an appropriate alternative to BMD for antifungal susceptibility testing of molds under specific circumstances, including testing with amphotericin B or triazoles for non-Aspergillus molds (Mucoromycotina and Fusarium spp.). Additional study of molecularly characterized triazole-resistant Aspergillus isolates is required to confirm the ability of the Etest method to detect voriconazole and posaconazole resistance among Aspergillus spp. PMID:26202113

  17. American Society of Clinical Oncology policy statement update: genetic testing for cancer susceptibility.

    PubMed

    2003-06-15

    As the leading organization representing cancer specialists involved in patient care and clinical research, the American Society of Clinical Oncology (ASCO) reaffirms its commitment to integrating cancer risk assessment and management, including molecular analysis of cancer predisposition genes, into the practice of oncology and preventive medicine. The primary goal of this effort is to foster expanded access to, and continued advances in, medical care provided to patients and families affected by hereditary cancer syndromes. The 1996 ASCO Statement on Genetic Testing for Cancer Susceptibility set forth specific recommendations relating to clinical practice, research needs, educational opportunities, requirement for informed consent, indications for genetic testing, regulation of laboratories, and protection from discrimination, as well as access to and reimbursement for cancer genetics services. In updating this Statement, ASCO endorses the following principles: Indications for Genetic Testing: ASCO recommends that genetic testing be offered when 1) the individual has personal or family history features suggestive of a genetic cancer susceptibility condition, 2) the test can be adequately interpreted, and 3) the results will aid in diagnosis or influence the medical or surgical management of the patient or family members at hereditary risk of cancer. ASCO recommends that genetic testing only be done in the setting of pre- and post-test counseling, which should include discussion of possible risks and benefits of cancer early detection and prevention modalities. Special Issues in Testing Children for Cancer Susceptibility: ASCO recommends that the decision to offer testing to potentially affected children should take into account the availability of evidence-based risk-reduction strategies and the probability of developing a malignancy during childhood. Where risk-reduction strategies are available or cancer predominantly develops in childhood, ASCO believes that

  18. Performance of the VITEK2 system for identification and susceptibility testing of routine Enterobacteriaceae clinical isolates.

    PubMed

    Pérez-Vázquez, M; Oliver, A; Sánchez del Saz, B; Loza, E; Baquero, F; Cantón, R

    2001-05-01

    The VITEK2 system was evaluated with 138 fresh consecutive routine clinical Enterobacteriaceae isolates. Susceptibility results to 10 beta-lactams, three aminoglycosides and a quinolone were compared with those obtained following the NCCLS standard microdilution. API20E was used as reference method for identification. All but three isolates were correctly identified in 3 h at species level (97.8%), two isolates (1.4%) at genus level and only one isolate was misidentified. Overall essential agreement for susceptibility testing was 97.1%. Discrepancies were mainly observed with piperacillin (1.1%), cefuroxime (0.6%) and amoxycillin/clavulanate (0.3%). Discrepancies for aminoglycosides and ciprofloxacin were low (<0.1%). Minor, major and very major errors (NCCLS categories) were 4.1%, 0.2% and 6.1%, respectively. Very major errors were due to piperacillin (4.5%), ampicillin (0.8%) and amoxycillin/clavulanate (0.8%). The VITEK2 system gave accurate identification and susceptibility testing results of routine Enterobacteriaceae clinical isolates. PMID:11337223

  19. Species Identification and Clarithromycin Susceptibility Testing of 278 Clinical Nontuberculosis Mycobacteria Isolates.

    PubMed

    Nie, Wenjuan; Duan, Hongfei; Huang, Hairong; Lu, Yu; Chu, Naihui

    2015-01-01

    Purpose of this paper is to analyze different species' proportion of nontuberculosis mycobacteria (NTM) and susceptibility to clarithromycin of different species. 278 clinical NTM isolates were identified into species by using 16S rRNA, rpoB and hsp65. Then clarithromycin susceptibility testing against different species was done separately, using microplate Alamar Blue assay. Finally, resistance isolates' erm(41) of M. abscessus were sequenced in order to analyze mechanisms for clarithromycin resistant. In this test, 131 isolates (47%) belonged to M. avium complex (MAC), and 70 isolates (25%) belonged to M. abscessus. Nearly all the M. abscessus subsp. abscessus resistant to clarithromycin had T28 in erm(41). However, all the M. abscessus subsp. abscessus susceptible to clarithromycin had C28 in erm(41). In this study, we find that MAC was the most common pathogens of NTM, and the second one was M. abscessus. However, M. chelonei, M. fuerth, and M. gordon were rare. Clarithromycin had a good inhibition activity against all the NTM species except M. abscessus subsp. abscessus. The erm(41) genotype is of high relevance to clarithromycin resistance. PMID:26146620

  20. Species Identification and Clarithromycin Susceptibility Testing of 278 Clinical Nontuberculosis Mycobacteria Isolates

    PubMed Central

    Nie, Wenjuan; Duan, Hongfei; Huang, Hairong; Lu, Yu; Chu, Naihui

    2015-01-01

    Purpose of this paper is to analyze different species' proportion of nontuberculosis mycobacteria (NTM) and susceptibility to clarithromycin of different species. 278 clinical NTM isolates were identified into species by using 16S rRNA, rpoB and hsp65. Then clarithromycin susceptibility testing against different species was done separately, using microplate Alamar Blue assay. Finally, resistance isolates' erm(41) of M. abscessus were sequenced in order to analyze mechanisms for clarithromycin resistant. In this test, 131 isolates (47%) belonged to M. avium complex (MAC), and 70 isolates (25%) belonged to M. abscessus. Nearly all the M. abscessus subsp. abscessus resistant to clarithromycin had T28 in erm(41). However, all the M. abscessus subsp. abscessus susceptible to clarithromycin had C28 in erm(41). In this study, we find that MAC was the most common pathogens of NTM, and the second one was M. abscessus. However, M. chelonei, M. fuerth, and M. gordon were rare. Clarithromycin had a good inhibition activity against all the NTM species except M. abscessus subsp. abscessus. The erm(41) genotype is of high relevance to clarithromycin resistance. PMID:26146620

  1. EUCAST recommendations for antimicrobial susceptibility testing applied to the three main Campylobacter species isolated in humans.

    PubMed

    Sifré, Elodie; Salha, Ben Amor; Ducournau, Astrid; Floch, Pauline; Chardon, Hubert; Mégraud, Francis; Lehours, Philippe

    2015-12-01

    Antimicrobial susceptibility testing of Campylobacter isolates is of great importance for treatment options especially in systemic diseases. The European Committee for Antimicrobial Susceptibility Testing (EUCAST) recently proposed epidemiological cut-offs (ECOFFs) for a limited number of antimicrobial compounds and for Campylobacter jejuni and Campylobacter coli only. In the present study, the EUCAST method was used after minor modifications to define antimicrobial susceptibility patterns for, 1997 C. jejuni, 419 C. coli and 100 Campylobacter fetus strains received at the French National Reference Center for Campylobacters and Helicobacters. Our results show that the ECOFFs defined by EUCAST for tetracycline and ciprofloxacin can be used for C. jejuni and C. coli. The same ECOFF can be used for erythromycin for the three species. The C. jejuni and C. coli ECOFFs for ciprofloxacin however cannot be applied to C. fetus. We also provide data to categorise two 2 β-lactams of interest for systemic diseases, ampicillin and amoxicillin+clavulanate, for the three species. PMID:26519770

  2. Multicenter evaluation of broth microdilution method for susceptibility testing of Cryptococcus neoformans against fluconazole.

    PubMed Central

    Sanati, H; Messer, S A; Pfaller, M; Witt, M; Larsen, R; Espinel-Ingroff, A; Ghannoum, M

    1996-01-01

    We have developed a microdilution method for measuring the susceptibility of Cryptococcus neoformans to fluconazole. The present study evaluated the interlaboratory agreement of the results for the microdilution method obtained at three different sites and compared this method with the National Committee for Clinical Laboratory Standards M27-P reference method. Excellent interlaboratory agreement among the results obtained at the three sites was achieved with this method (83 and 96% agreement within 1 and 2 log2 dilutions, respectively). An overall agreement of 90% between the microdilution method and the M27-P method was observed, demonstrating the comparability of the two methods. However, there are inherent problems with the M27-P method in relation to measuring C. neoformans susceptibility, including suboptimal growth of the organism in RPMI 1640, a longer incubation period, and a narrow range of MICs. On the basis of these data, the microdilution method tested in this study is recommended for inclusion in the National Committee for Laboratory Standards method for testing the antifungal susceptibility of C. neoformans. PMID:8727919

  3. An investigation of tests of susceptibility to noise-induced hearing loss

    NASA Astrophysics Data System (ADS)

    Lawton, B. W.; Robinson, D. W.

    1986-08-01

    Susceptibility to hearing damage was studied by testing 81 males aged 16 to 27 yr, and 50 males aged 45 to 65 yr. The older men were then screened for better than average hearing in an attempt to identify an innate factor protecting them from noise damage. Results suggest that there is no interaction between the psychoacoustical and neuromechanical properties of the ear. Cochlear function seems to be independent from the reflex self-protection of the ear. However, outliers of the variate distributions, i.e., robust or susceptible ears, were further analyzed. Correlations of rank, free from artefacts imposed by the physical scales of measurement, identified a small number of consistent outliers, and demonstrate the statistical independence of three properties of human hearing: conductive efficiency, cochlear function, and reflex function.

  4. Antimicrobial Disk Susceptibility Testing of Leptospira spp. Using Leptospira Vanaporn Wuthiekanun (LVW) Agar.

    PubMed

    Wuthiekanun, Vanaporn; Amornchai, Premjit; Langla, Sayan; White, Nicholas J; Day, Nicholas P J; Limmathurotsakul, Direk; Peacock, Sharon J

    2015-08-01

    Leptospira Vanaporn Wuthiekanun (LVW) agar was used to develop a disk diffusion assay for Leptospira spp. Ten pathogenic Leptospira isolates were tested, all of which were susceptible to 17 antimicrobial agents (amoxicillin/clavulanic acid, amoxicillin, azithromycin, cefoxitin, ceftazidime, ceftriaxone, chloramphenicol, ciprofloxacin, clindamycin, doripenem, doxycycline, gentamicin, linezolid, nitrofurantoin, penicillin, piperacillin/tazobactam, and tetracycline). All 10 isolates had no zone of growth inhibition for four antimicrobials (fosfomycin, nalidixic acid, rifampicin, and trimethoprim/sulfamethoxazole). Of the ten Leptospira, seven had a growth inhibition zone of ≤ 21 mm for aztreonam, the zone diameter susceptibility break point for Enterobacteriaceae. This assay could find utility as a simple screening method during the epidemiological surveillance of antimicrobial resistance in Leptospira spp. PMID:26055750

  5. Antimicrobial Susceptibility Test with Plasmonic Imaging and Tracking of Single Bacterial Motions on Nanometer Scale.

    PubMed

    Syal, Karan; Iriya, Rafael; Yang, Yunze; Yu, Hui; Wang, Shaopeng; Haydel, Shelley E; Chen, Hong-Yuan; Tao, Nongjian

    2016-01-26

    Antimicrobial susceptibility tests (ASTs) are important for confirming susceptibility to empirical antibiotics and detecting resistance in bacterial isolates. Currently, most ASTs performed in clinical microbiology laboratories are based on bacterial culturing, which take days to complete for slowly growing microorganisms. A faster AST will reduce morbidity and mortality rates and help healthcare providers administer narrow spectrum antibiotics at the earliest possible treatment stage. We report the development of a nonculture-based AST using a plasmonic imaging and tracking (PIT) technology. We track the motion of individual bacterial cells tethered to a surface with nanometer (nm) precision and correlate the phenotypic motion with bacterial metabolism and antibiotic action. We show that antibiotic action significantly slows down bacterial motion, which can be quantified for development of a rapid phenotypic-based AST. PMID:26637243

  6. Activity of fosfomycin and comparison of several susceptibility testing methods against contemporary urine isolates.

    PubMed

    Hirsch, Elizabeth B; Raux, Brian R; Zucchi, Paola C; Kim, Yisu; McCoy, Christopher; Kirby, James E; Wright, Sharon B; Eliopoulos, George M

    2015-12-01

    Fosfomycin is recommended as first-line treatment for acute uncomplicated cystitis in women. It has demonstrated in vitro activity against a variety of pathogens; however, a paucity of data are available from the USA. We determined the susceptibility of a collection of urine isolates to fosfomycin and compared multiple methods of susceptibility testing. Consecutive non-duplicate Enterobacteriaceae, enterococci and Pseudomonas aeruginosa isolates were collected from the clinical microbiology laboratory between August 2013 and January 2014. Isolates represented hospitalised or emergency department patients with monomicrobial bacteriuria. Fosfomycin MICs were determined in duplicate, on separate days, by Etest and disk diffusion and results were compared with agar dilution. Nitrofurantoin and ciprofloxacin were used as comparators. MIC results were categorised using Clinical and Laboratory Standards Institute interpretive criteria for Escherichia coli and Enterococcus faecalis. Correlation between the three testing methods was evaluated. Overall susceptibility to fosfomycin was 94.4%, 93.5% and 87.9% by agar dilution, disk diffusion and Etest, respectively. Five fosfomycin-resistant isolates were identified, including two Morganella morganii, one P. aeruginosa, one Proteus mirabilis and one Enterobacter aerogenes. Across all organisms, rates of essential agreement, categorical agreement, minor errors, major errors and very major errors for Etest/disk diffusion compared with agar dilution were 77.3%/NA, 89.5/93.8%, 7.1/5.0%, 3.6/1.3% and 0/0%, respectively. Fosfomycin displayed fairly consistent activity against a majority of isolates collected when using the susceptibility breakpoint of 64 μg/mL. MICs for E. coli were particularly low (≤2 μg/mL). These data lend support to current guidelines that recommend fosfomycin as empirical first-line therapy for uncomplicated UTI. PMID:26498988

  7. Beyond Testing: Assessment for Teaching and Learning. Viewpoints. [Booklet and Audio Compact Discs].

    ERIC Educational Resources Information Center

    North Central Regional Educational Lab., Naperville, IL.

    This multimedia package, which contains two audio CDs and a short informative booklet, examines the use of assessment to improve teaching and learning. The booklet contains the essay "Assessment-Driven Reform: A Leadership Approach" (Rhetta L. Detrich with Ed Janus and Sabrina W. M. Laine). It discusses the increasing importance of testing and…

  8. Variability of β-lactam susceptibility testing for Streptococcus pneumoniae using 4 commercial test methods and broth microdilution.

    PubMed

    Charles, Marthe K; Berenger, Byron M; Turnbull, LeeAnn; Rennie, Robert; Fuller, Jeff

    2016-03-01

    Limited data are available that verify the performance of commercial susceptibility methods for Streptococcus pneumoniae following the 2008 Clinical and Laboratory Standards Institute revision of the β-lactam breakpoints. We compared the performance of Etest, M.I.C. Evaluator (M.I.C.E), Vitek, and Sensititre systems to broth microdilution for S. pneumoniae susceptibility testing of penicillin, ceftriaxone, meropenem, and amoxicillin. Essential agreement was ≥90% for the majority of the β-lactams and methods tested, particularly for penicillin and ceftriaxone. Categorical agreements (CAs) for penicillin using meningeal and nonmeningeal breakpoints were ≥90%; CAs using penicillin oral breakpoints were 84-89%. Ceftriaxone CAs using nonmeningeal and meningeal breakpoints were 68-88% for Etest, M.I.C.E., and Vitek2 with 6-12% very major errors (VMEs) using meningeal breakpoints. Sensititre CAs for ceftriaxone, amoxicillin, and meropenem were ≥90% with no VMEs. In the context of the current guidelines, there exists considerable method-dependent variability in the susceptibility of S. pneumoniae to β-lactams. PMID:26707068

  9. Use of in vitro vancomycin testing results to predict susceptibility to oritavancin, a new long-acting lipoglycopeptide.

    PubMed

    Jones, Ronald N; Turnidge, John D; Moeck, Greg; Arhin, Francis F; Mendes, Rodrigo E

    2015-04-01

    Oritavancin is a recently approved lipoglycopeptide antimicrobial agent with activity against Gram-positive pathogens. Its extended serum elimination half-life and concentration-dependent killing enable single-dose treatment of acute bacterial skin and skin structure infections. At the time of regulatory approval, new agents, including oritavancin, are not offered in the most widely used susceptibility testing devices and therefore may require application of surrogate testing using a related antimicrobial to infer susceptibility. To evaluate vancomycin as a predictive susceptibility marker for oritavancin, 26,993 recent Gram-positive organisms from U.S. and European hospitals were tested using reference MIC methods. Organisms included Staphylococcus aureus, coagulase-negative staphylococci (CoNS), beta-hemolytic streptococci (BHS), viridans group streptococci (VGS), and enterococci (ENT). These five major pathogen groups were analyzed by comparing results with FDA-approved susceptible breakpoints for both drugs, as well as those suggested by epidemiological cutoff values and supported by pharmacokinetic/pharmacodynamic analyses. Vancomycin susceptibility was highly accurate (98.1 to 100.0%) as a surrogate for oritavancin susceptibility among the indicated pathogen species. Furthermore, direct MIC comparisons showed high oritavancin potencies, with vancomycin/oritavancin MIC90 results of 1/0.06, 2/0.06, 0.5/0.12,1/0.06, and >16/0.06 μg/ml for S. aureus, CoNS, BHS, VGS, and ENT, respectively. In conclusion, vancomycin demonstrated acceptable accuracy as a surrogate marker for predicting oritavancin susceptibility when tested against the indicated pathogens. In contrast, 93.3% of vancomycin-nonsusceptible enterococci had oritavancin MIC values of ≤0.12 μg/ml, indicating a poor predictive value of vancomycin for oritavancin resistance against these organisms. Until commercial oritavancin susceptibility testing devices are readily available, isolates that when

  10. A Rapid Molecular Test for Determining Yersinia pestis Susceptibility to Ciprofloxacin by the Quantification of Differentially Expressed Marker Genes.

    PubMed

    Steinberger-Levy, Ida; Shifman, Ohad; Zvi, Anat; Ariel, Naomi; Beth-Din, Adi; Israeli, Ofir; Gur, David; Aftalion, Moshe; Maoz, Sharon; Ber, Raphael

    2016-01-01

    Standard antimicrobial susceptibility tests used to determine bacterial susceptibility to antibiotics are growth dependent and time consuming. The long incubation time required for standard tests may render susceptibility results irrelevant, particularly for patients infected with lethal bacteria that are slow growing on agar but progress rapidly in vivo, such as Yersinia pestis. Here, we present an alternative approach for the rapid determination of antimicrobial susceptibility, based on the quantification of the changes in the expression levels of specific marker genes following exposure to growth-inhibiting concentrations of the antibiotic, using Y. pestis and ciprofloxacin as a model. The marker genes were identified by transcriptomic DNA microarray analysis of the virulent Y. pestis Kimberley53 strain after exposure to specific concentrations of ciprofloxacin for various time periods. We identified several marker genes that were induced following exposure to growth-inhibitory concentrations of ciprofloxacin, and we confirmed the marker expression profiles at additional ciprofloxacin concentrations using quantitative RT-PCR. Eleven candidate marker transcripts were identified, of which four mRNA markers were selected for a rapid quantitative RT-PCR susceptibility test that correctly determined the Minimal Inhibitory Concentration (MIC) values and the categories of susceptibility of several Y. pestis strains and isolates harboring various ciprofloxacin MIC values. The novel molecular susceptibility test requires just 2 h of antibiotic exposure in a 7-h overall test time, in contrast to the 24 h of antibiotic exposure required for a standard microdilution test. PMID:27242774

  11. A Rapid Molecular Test for Determining Yersinia pestis Susceptibility to Ciprofloxacin by the Quantification of Differentially Expressed Marker Genes

    PubMed Central

    Steinberger-Levy, Ida; Shifman, Ohad; Zvi, Anat; Ariel, Naomi; Beth-Din, Adi; Israeli, Ofir; Gur, David; Aftalion, Moshe; Maoz, Sharon; Ber, Raphael

    2016-01-01

    Standard antimicrobial susceptibility tests used to determine bacterial susceptibility to antibiotics are growth dependent and time consuming. The long incubation time required for standard tests may render susceptibility results irrelevant, particularly for patients infected with lethal bacteria that are slow growing on agar but progress rapidly in vivo, such as Yersinia pestis. Here, we present an alternative approach for the rapid determination of antimicrobial susceptibility, based on the quantification of the changes in the expression levels of specific marker genes following exposure to growth-inhibiting concentrations of the antibiotic, using Y. pestis and ciprofloxacin as a model. The marker genes were identified by transcriptomic DNA microarray analysis of the virulent Y. pestis Kimberley53 strain after exposure to specific concentrations of ciprofloxacin for various time periods. We identified several marker genes that were induced following exposure to growth-inhibitory concentrations of ciprofloxacin, and we confirmed the marker expression profiles at additional ciprofloxacin concentrations using quantitative RT-PCR. Eleven candidate marker transcripts were identified, of which four mRNA markers were selected for a rapid quantitative RT-PCR susceptibility test that correctly determined the Minimal Inhibitory Concentration (MIC) values and the categories of susceptibility of several Y. pestis strains and isolates harboring various ciprofloxacin MIC values. The novel molecular susceptibility test requires just 2 h of antibiotic exposure in a 7-h overall test time, in contrast to the 24 h of antibiotic exposure required for a standard microdilution test. PMID:27242774

  12. Reverberation Chamber Uniformity Validation and Radiated Susceptibility Test Procedures for the NASA High Intensity Radiated Fields Laboratory

    NASA Technical Reports Server (NTRS)

    Koppen, Sandra V.; Nguyen, Truong X.; Mielnik, John J.

    2010-01-01

    The NASA Langley Research Center's High Intensity Radiated Fields Laboratory has developed a capability based on the RTCA/DO-160F Section 20 guidelines for radiated electromagnetic susceptibility testing in reverberation chambers. Phase 1 of the test procedure utilizes mode-tuned stirrer techniques and E-field probe measurements to validate chamber uniformity, determines chamber loading effects, and defines a radiated susceptibility test process. The test procedure is segmented into numbered operations that are largely software controlled. This document is intended as a laboratory test reference and includes diagrams of test setups, equipment lists, as well as test results and analysis. Phase 2 of development is discussed.

  13. Assessment of Etest as an alternative to agar dilution for antimicrobial susceptibility testing of Neisseria gonorrhoeae.

    PubMed

    Liu, Hsi; Taylor, Thomas H; Pettus, Kevin; Trees, David

    2014-05-01

    We studied whether the Etest can be used as an alternative to agar dilution to determine antimicrobial susceptibilities of ceftriaxone, cefixime, and cefpodoxime in Neisseria gonorrhoeae surveillance. One hundred fifteen clinical and laboratory isolates of N. gonorrhoeae were tested following the Clinical Laboratory Improvement Amendments (CLIA)-approved CLSI standard agar dilution method and, separately, by the Etest according to the manufacturer's recommendations. The MICs were determined and compared. Ten laboratory-generated mutants were used to simulate substantially nonsusceptible specimens. The Etest and agar dilution methods were well correlated. Statistical tests produced regression R2 values of 88%, 82%, and 85% and Pearson correlation coefficients of 92%, 91%, and 92% for ceftriaxone, cefixime, and cefpodoxime, respectively. When paired comparisons were made, the two tests were 88.7%, 80%, and 87% within 1 log2 dilution from each other for ceftriaxone, cefixime, and cefpodoxime, respectively. The within-2-log2 agreements were 99.1%, 98.3%, and 94.8% for ceftriaxone, cefixime, and cefpodoxime, respectively. Notwithstanding the good correlations and the within-2-log2 general agreement, the Etest results produced slightly lower MICs than the agar dilution results. In conclusion, we found that the Etest can be effectively used as an alternative to agar dilution testing to determine the susceptibility of N. gonorrhoeae to ceftriaxone, cefixime, and cefpodoxime, although we recommend further research into extremely resistant isolates. For isolates within the typical range of clinical MICs, reexamination of the Etest interpretation of susceptible and nonsusceptible categories would likely allow for successful transition from agar dilution to the Etest. PMID:24554750

  14. Assessment of Etest as an Alternative to Agar Dilution for Antimicrobial Susceptibility Testing of Neisseria gonorrhoeae

    PubMed Central

    Taylor, Thomas H.; Pettus, Kevin; Trees, David

    2014-01-01

    We studied whether the Etest can be used as an alternative to agar dilution to determine antimicrobial susceptibilities of ceftriaxone, cefixime, and cefpodoxime in Neisseria gonorrhoeae surveillance. One hundred fifteen clinical and laboratory isolates of N. gonorrhoeae were tested following the Clinical Laboratory Improvement Amendments (CLIA)-approved CLSI standard agar dilution method and, separately, by the Etest according to the manufacturer's recommendations. The MICs were determined and compared. Ten laboratory-generated mutants were used to simulate substantially nonsusceptible specimens. The Etest and agar dilution methods were well correlated. Statistical tests produced regression R2 values of 88%, 82%, and 85% and Pearson correlation coefficients of 92%, 91%, and 92% for ceftriaxone, cefixime, and cefpodoxime, respectively. When paired comparisons were made, the two tests were 88.7%, 80%, and 87% within 1 log2 dilution from each other for ceftriaxone, cefixime, and cefpodoxime, respectively. The within-2-log2 agreements were 99.1%, 98.3%, and 94.8% for ceftriaxone, cefixime, and cefpodoxime, respectively. Notwithstanding the good correlations and the within-2-log2 general agreement, the Etest results produced slightly lower MICs than the agar dilution results. In conclusion, we found that the Etest can be effectively used as an alternative to agar dilution testing to determine the susceptibility of N. gonorrhoeae to ceftriaxone, cefixime, and cefpodoxime, although we recommend further research into extremely resistant isolates. For isolates within the typical range of clinical MICs, reexamination of the Etest interpretation of susceptible and nonsusceptible categories would likely allow for successful transition from agar dilution to the Etest. PMID:24554750

  15. The effect of age on result of straight leg raising test in patients suffering lumbar disc herniation and sciatica

    PubMed Central

    Tabesh, Homayoun; Tabesh, Ariyan; Fakharian, Esmaeil; Fazel, Mohammadreza; Abrishamkar, Saeid

    2015-01-01

    Background: Ninety percent of all people sometimes during their lives experience low back pain, and 30-40% develops radicular leg pain with the sciatica characteristics. Although for clinical diagnosis of lumbar disc herniation (LDH) straight leg raising (SLR) test in 85-90% of cases indicates LDH, but in our practice with LDH patients this test is frequently negative despite radicular leg pain due to LDH. Hence, we decided to evaluate this test in LDH in different age groups. Materials and Methods: All patients with leg pain referring to neurosurgery clinic were enrolled. Those with a history of pain other than sciatica excluded and SLR test and magnetic resonance imaging (MRI) of the lumbosacral spine performed. The patients with negative MRI findings excluded and finally 269 patients with true sciatica and positive MRI were included. SLR tests were performed for different age groups. Results: Of 269 patients, 167 were male. The age range was 16-80 years. The most involved levels were L5-S1 (47%) and L4-L5 (42%), respectively. The rate of positive SLR result, which was 100%, 87% and 82% for 10-19, 20-29 and 30-39 years age group respectively. With an increment of age, the rate of positive test regularly declined. The chance of positive SLR in men is 1.3 times the women (odds ratio [OR] 2.4; 95% confidence interval [CI] = 1.265-4.557; P = 0.007). Increasing the age has suppression effect in positivity of SLR so that for each 1-year the chance of SLR become 0.27 times less to become positive and this is also statically meaningful (OR = 0.271;95% CI = 0.188-0.391; P,0.001). The chance of positive SLR for patients under 60 is 5.4 folds more than patients above 60 years old (OR = 5.4; 95% CI = 4-8.3; P, 0.001). Conclusion: Age, sex (male), and disk level had statistically the effect on SLR positive test. PMID:25983767

  16. Termite-Susceptible Species of Wood for Inclusion as a Reference in Indonesian Standardized Laboratory Testing.

    PubMed

    Arinana; Tsunoda, Kunio; Herliyana, Elis N; Hadi, Yusuf S

    2012-01-01

    Standardized laboratory testing of wood and wood-based products against subterranean termites in Indonesia (SNI 01.7207-2006) (SNI) has no requirement for the inclusion of a comparative reference species of wood (reference control). This is considered a weakness of the Indonesian standard. Consequently, a study was undertaken to identify a suitable Indonesian species of community wood that could be used as a reference control. Four candidate species of community woods: Acacia mangium, Hevea brasiliensis, Paraserianthes falcataria and Pinus merkusii were selected for testing their susceptibility to feeding by Coptotermes formosanus. Two testing methods (SNI and the Japanese standard method JIS K 1571-2004) were used to compare the susceptibility of each species of wood. Included in the study was Cryptomeria japonica, the reference control specified in the Japanese standard. The results of the study indicated that P. merkusii is a suitable reference species of wood for inclusion in laboratory tests against subterranean termites, conducted in accordance with the Indonesian standard (SNI 01.7207-2006). PMID:26466532

  17. Direct Susceptibility Testing of Mycobacterium tuberculosis for Pyrazinamide by Use of the Bactec MGIT 960 System

    PubMed Central

    Demers, Anne-Marie; Venter, Amour; Friedrich, Sven O.; Rojas-Ponce, Gabriel; Mapamba, Daniel; Jugheli, Levan; Sasamalo, Mohammed; Almeida, Deepak; Dorasamy, Afton; Jentsch, Ute; Gibson, Mara; Everitt, Daniel; Diacon, Andreas H.

    2016-01-01

    Pyrazinamide (PZA) is a key antituberculosis drug, yet no rapid susceptibility test is commercially available. PZA drug susceptibility testing (DST) was performed directly on sputum samples from 327 patients and compared with the indirect method by using the Bactec MGIT 960 system in the context of patient screening for participation in a drug trial. Compared to standard indirect PZA DST, direct DST was successful in only 59% of cases, but results obtained were highly accurate and available faster. Agreement between the direct and indirect methods varied from 90 to 100% in each laboratory. The median times for obtaining PZA results from the time when the specimen was collected ranged from 11 to 16 days for the direct test and 18 to 95 days for the indirect test across laboratories. The direct method is accurate and reproducible across laboratories. It can be expected to accelerate results in >50% of cases, but it cannot replace indirect DST for PZA. Phenotypic methods remain the gold standard for DST in drug trials. If future studies can optimize the method to decrease the number of uninterpretable results, direct MGIT DST could be the new phenotypic DST standard for clinical trials, providing more rapid detection of resistance to new drugs in experimental regimens. PMID:26912751

  18. Termite-Susceptible Species of Wood for Inclusion as a Reference in Indonesian Standardized Laboratory Testing

    PubMed Central

    Arinana; Tsunoda, Kunio; Herliyana, Elis N.; Hadi, Yusuf S.

    2012-01-01

    Standardized laboratory testing of wood and wood-based products against subterranean termites in Indonesia (SNI 01.7207-2006) (SNI) has no requirement for the inclusion of a comparative reference species of wood (reference control). This is considered a weakness of the Indonesian standard. Consequently, a study was undertaken to identify a suitable Indonesian species of community wood that could be used as a reference control. Four candidate species of community woods: Acacia mangium, Hevea brasiliensis, Paraserianthes falcataria and Pinus merkusii were selected for testing their susceptibility to feeding by Coptotermes formosanus. Two testing methods (SNI and the Japanese standard method JIS K 1571-2004) were used to compare the susceptibility of each species of wood. Included in the study was Cryptomeria japonica, the reference control specified in the Japanese standard. The results of the study indicated that P. merkusii is a suitable reference species of wood for inclusion in laboratory tests against subterranean termites, conducted in accordance with the Indonesian standard (SNI 01.7207-2006). PMID:26466532

  19. Rapid susceptibility testing of Mycobacterium tuberculosis by bioluminescence assay of mycobacterial ATP

    SciTech Connect

    Nilsson, L.E.; Hoffner, S.E.; Ansehn, S.

    1988-08-01

    Mycobacterial growth was monitored by bioluminescence assay of mycobacterial ATP. Cultures of Mycobacterium tuberculosis H37Rv and of 25 clinical isolates of the same species were exposed to serial dilutions of ethambutol, isoniazid, rifampin, and streptomycin. A suppression of ATP, indicating growth inhibition, occurred for susceptible but not resistant strains within 5 to 7 days of incubation. Breakpoint concentrations between susceptibility and resistance were determined by comparing these results with those obtained by reference techniques. Full agreement was found in 99% of the assays with the resistance ratio method on Lowenstein-Jensen medium, and 98% of the assays were in full agreement with the radiometric system (BACTEC). A main advantage of the bioluminescence method is its rapidity, with results available as fast as with the radiometric system but at a lower cost and without the need for radioactive culture medium. The method provides kinetic data concerning drug effects within available in vivo drug concentrations and has great potential for both rapid routine susceptibility testing and research applications in studies of drug effects on mycobacteria.

  20. Antimicrobial susceptibility testing of Brachyspira intermedia and Brachyspira pilosicoli isolates from Australian chickens.

    PubMed

    Hampson, D J; Stephens, C P; Oxberry, S L

    2006-02-01

    Susceptibilities of predominantly Australian isolates of the pathogenic intestinal spirochaetes Brachyspira intermedia (n = 25) and Brachyspira pilosicoli (n = 17) from chickens were tested in agar dilution against four concentrations each of the antimicrobials tiamulin, lincomycin, tylosin, metronidazole, tetracycline and ampicillin. Based on available minimum inhibitory concentration (MIC) breakpoint values for Brachyspira hyodysenteriae or other Gram-negative enteric veterinary pathogens, isolates of both species generally were susceptible to tiamulin, lincomycin, metronidazole and tetracycline. Although not classed as resistant, four isolates of B. intermedia had an elevated MIC range for tiamulin (1 to 4 mg/l), 11 isolates of B. intermedia and five of B. pilosicoli had an elevated MIC range for lincomycin (10 to 50 mg/l), one isolate of B. pilosicoli had an elevated MIC range for tetracycline (10 to 20 mg/l), and one isolate of B. intermedia and five of B. pilosicoli had an elevated MIC range for ampicillin (10 to 50 mg/l). A clear lack of susceptibility to tylosin (MIC > 4 mg/l) was seen in 11 isolates each of B. intermedia and B. pilosicoli, and to ampicillin (MIC > 32 mg/l) in two isolates of B. pilosicoli. These data suggest that some resistance to common antimicrobials exists among intestinal spirochetes obtained from laying hens and supports the need of MIC data for clinical isolates before any treatment is considered. PMID:16448937

  1. Genetic antimicrobial susceptibility testing in Gram-negative sepsis - impact on time to results in a routine laboratory.

    PubMed

    Kommedal, Øyvind; Aasen, Johanne Lind; Lindemann, Paul Christoffer

    2016-07-01

    Diagnostic testing of positive blood cultures is among the most critical tasks performed by clinical microbiology laboratories, and the total analysis time from sampling to results should be kept as short as possible. By providing identification of pelleted bacteria directly from positive blood-cultures, MALDI-TOF MS opens for relatively low-complex species-adjusted genetic susceptibility testing from the same bacterial pellet. In our lab routine, we prospectively evaluated a rapid in-house real-time PCR targeting the most common aminoglycoside and cephalosporin resistance genes in Escherichia coli and Klebsiella pneumoniae and measured time to preliminary susceptibility reporting for 138 samples. The results were compared to direct phenotypic susceptibility testing with interpretation after 6 h and overnight incubation respectively. Results from the genetic susceptibility testing were available for 69.5% (96/138) of the positive blood cultures within 24 h after sample collection. No phenotypic susceptibility results were available at this time. Compared to overnight direct susceptibility testing, the average time from sample collection to preliminary susceptibility reporting was reduced with 43%, from 45 h and 5 min to 25 h and 44 min, providing an earlier adjustment of antimicrobial therapy for 12 patients. Minor logistic adjustments have the potential to save yet another 4 h. PMID:27197792

  2. CLSI performance standards for antimicrobial susceptibility testing of bacteria isoloated from aquatic animals; second information supplement. CLSI document VET03/VET04-S2

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The supplemental information presented in this document is intended for use with the antimicrobial susceptibility testing procedures published in the following Clinical and Laboratory Standards Institute (CLSI) approved documents VET03-A Methods for Antimicrobial Disk Susceptibility Testing of Bacte...

  3. Preparation of ormetoprim-sulfadimethoxine-medicated discs for disc diffusion assay

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Romet ( a blend of ormetoprim and sulfadimethoxine) is a type A medicated article for the manufacture of medicated feed in the catfish industry. Recently, the commercial manufacture of ormetoprim-sulfadimethoxine susceptibility discs was discontinued. Ormetoprim-sulfadimethoxine discs were prepare...

  4. Direct disk diffusion test using European Clinical Antimicrobial Susceptibility Testing breakpoints provides reliable results compared with the standard method

    PubMed Central

    Stokkou, Sofia; Geginat, Gernot; Schlüter, Dirk

    2015-01-01

    Sepsis represents a life-threatening infection requiring the immediate start of antibacterial treatment to reduce morbidity. Thus, laboratories use direct antimicrobial susceptibility testing (AST) to rapidly generate preliminary results from positive blood cultures. As the direct AST has not yet been published to be evaluated with EUCAST breakpoints, the purpose of the study was to investigate the reliability of the direct agar diffusion test to correctly produce AST results from positive monobacterial blood cultures compared with the VITEK2-based definitive AST, when current EUCAST breakpoints were used. A total of 428 isolates from unselected monobacterial routine blood cultures and 110 challenge strains were included. Direct agar diffusion-based and standard VITEK2-based AST of 2803 bacterium–drug combinations yielded a total clinical category agreement of 95.47% with 1.28% very major errors and 3.42% combined major and minor errors. On the species level, very major errors were observed in the species–drug combinations Enterococcus spp.–high-level gentamicin (10.87%) and Staphylococcus spp.–rifampicin (5%), only. No very major errors occurred with Enterobacteriaceae and Pseudomonas aeruginosa. In most species–drug combinations, the direct agar diffusion test using EUCAST breakpoints precisely predicted the result of the definitive antibiotic susceptibility test and, thus, it can be used to optimize empiric antibiotic therapy until definitive results are available. PMID:25883798

  5. Direct disk diffusion test using European Clinical Antimicrobial Susceptibility Testing breakpoints provides reliable results compared with the standard method.

    PubMed

    Stokkou, Sofia; Geginat, Gernot; Schlüter, Dirk; Tammer, Ina

    2015-03-01

    Sepsis represents a life-threatening infection requiring the immediate start of antibacterial treatment to reduce morbidity. Thus, laboratories use direct antimicrobial susceptibility testing (AST) to rapidly generate preliminary results from positive blood cultures. As the direct AST has not yet been published to be evaluated with EUCAST breakpoints, the purpose of the study was to investigate the reliability of the direct agar diffusion test to correctly produce AST results from positive monobacterial blood cultures compared with the VITEK2-based definitive AST, when current EUCAST breakpoints were used. A total of 428 isolates from unselected monobacterial routine blood cultures and 110 challenge strains were included. Direct agar diffusion-based and standard VITEK2-based AST of 2803 bacterium-drug combinations yielded a total clinical category agreement of 95.47% with 1.28% very major errors and 3.42% combined major and minor errors. On the species level, very major errors were observed in the species-drug combinations Enterococcus spp.-high-level gentamicin (10.87%) and Staphylococcus spp.-rifampicin (5%), only. No very major errors occurred with Enterobacteriaceae and Pseudomonas aeruginosa. In most species-drug combinations, the direct agar diffusion test using EUCAST breakpoints precisely predicted the result of the definitive antibiotic susceptibility test and, thus, it can be used to optimize empiric antibiotic therapy until definitive results are available. PMID:25883798

  6. [Susceptibilities of clinical bacterial isolates to antimicrobial agents. A study mainly focused on imipenem. Reported by the Research Group for Testing Imipenem Susceptibility on Clinical Isolates].

    PubMed

    Igari, J

    1990-11-01

    This study was conducted to investigate susceptibilities of clinical bacterial isolates to imipenem (IPM) and other antibacterial agents at 64 hospital laboratories throughout Japan from September to December of 1988. In this study, identification and susceptibility testing were carried out at each laboratory and the tests were performed according to the disk dilution method recommended by NCCLS in which susceptibilities are classified into "S", "MS", "I" and "R". IPM showed markedly high in vitro activities against Streptococcus pneumoniae, Streptococcus pyogenes, Streptococcus agalactiae, Enterococcus faecalis, Haemophilus influenzae, Escherichia coli, Klebsiella pneumoniae, Klebsiella oxytoca, Enterobacter aerogenes, Enterobacter cloacae, Serratia marcescens, Salmonella spp., Citrobacter freundii, Proteus mirabilis, Proteus vulgaris, Morganella morganii, Providencia rettgeri, Providencia stuartii, Acinetobacter calcoaceticus, Moraxella (Branhamella) catarrhalis, Alcaligenes spp., Peptococcus spp./Peptostreptococcus spp., Bacteroides fragilis and Bacteroides spp. IPM also had strong activities against Achromobacter xylosoxidans and Pseudomonas aeruginosa, but less active against Flavobacterium spp., E. faecium, coagulase-negative staphylococci (CNS), Staphylococcus aureus and Pseudomonas cepacia. In a study in which activities of IPM against bacteria isolated from different clinical sources were compared, differences in susceptibilities were observed among S. aureus, CNS, A. calcoaceticus and P. aeruginosa, but such differences were not apparent among S. pneumoniae, E. faecalis, H. influenzae, E. coli, K. pneumoniae, E. cloacae, C. freundii, S. marcescens or P. mirabilis. PMID:2287060

  7. Update from the Laboratory: Clinical Identification and Susceptibility Testing of Fungi and Trends in Antifungal Resistance.

    PubMed

    Albataineh, Mohammad T; Sutton, Deanna A; Fothergill, Annette W; Wiederhold, Nathan P

    2016-03-01

    Despite the availability of new diagnostic assays and broad-spectrum antifungal agents, invasive fungal infections remain a significant challenge to clinicians and are associated with marked morbidity and mortality. In addition, the number of etiologic agents of invasive mycoses has increased accompanied by an expansion in the immunocompromised patient populations, and the use of molecular tools for fungal identification and characterization has resulted in the discovery of several cryptic species. This article reviews various methods used to identify fungi and perform antifungal susceptibility testing in the clinical laboratory. Recent developments in antifungal resistance are also discussed. PMID:26739605

  8. Accuracy of different methods for susceptibility testing of gentamicin with KPC carbapenemase-producing Klebsiella pneumoniae.

    PubMed

    Arena, Fabio; Giani, Tommaso; Vaggelli, Guendalina; Terenzi, Giovanni; Pecile, Patrizia; Rossolini, Gian Maria

    2015-02-01

    Performance of Vitek2, Etest, and TREK broth microdilution (BMD) panels was evaluated versus reference CLSI BMD for gentamicin susceptibility testing with 57 bloodstream isolates of KPC-producing Klebsiella pneumoniae. Compared with reference BMD, the Essential Agreement and Categorical Agreement for TREK panels, Vitek2, and Etest were 91.2%, 31.6%, and 61.4%, respectively, and 86%, 21%, and 52.6%, respectively. Four very major discrepancies occurred with Vitek2. In these 4 strains, gentamicin resistance was associated with the presence of an armA aminoglycoside resistance determinant. PMID:25533616

  9. In vitro susceptibility testing of nonsporing anaerobes to ten antimicrobial agents.

    PubMed

    Rao, P S; Shivananda, P G

    2000-07-01

    Antibiotic susceptibility was performed on sixty clinical isolates of nonsporing anaerobes against ten antimicrobial agents. The test was performed on Muller Hinton Agar and Wilkins Chalgren blood agar by preparing suspension of freshly isolated colonies in BHI broth. Apart from Metronidazole and Chloramphenicol newer antibiotics like Minocycline, Secnidazole, Tinidazole, Clarithromycin, Roxithromycin were also tried. Antimicrobial agents like Metronidazole, Secnidazole, Tinidazole and Minocycline were 100% sensitive, followed by Chloramphenicol, Clarithromycin and Roxithromycin. These newer agents can be good alternatives for the treatment of non sporing anaerobes. PMID:11218673

  10. In vitro susceptibility testing of nonsporing anaerobes to ten antimicrobial agents.

    PubMed

    Rao, P S; Shivananda, P G

    2000-04-01

    Antibiotic Susceptibility was performed on sixty clinical isolates of nonsporing anaerobes against ten antimicrobial agents. The test was performed on Muller Hinton Agar and Wilkins Chalgren blood agar by preparing suspension of freshly isolated colonies in BHI broth. Apart from Metronidazole and Chloramphenicol newer antibiotics like Minocycline, Secnidazole, Tinidazole, Clarithromycin, Roxithromycin were also tried. Antimicrobial agents like metronidazole, Secnidazole, Tinidazole and Minocycline were 100% sensitive, followed by Chloramphenicol, Clarithromycin and Roxithromycin. These newer agents can be good alternatives for the treatment of non sporing anaerobes. PMID:11217270

  11. Methods for broth dilution susceptibility testing of bacteria isolated from aquatic animals; approved guideline-second edition

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Antimicrobial susceptibility testing is recommended to determine which antimicrobial agents should be considered for treating a bacterial pathogen. Many bacteria that cause disease in aquatic animals require growth conditions that vary substantially from routine terrestrial pathogens. It has thus ...

  12. Evaluating Constraints on Heavy-Ion SEE Susceptibility Imposed by Proton SEE Testing and Other Mixed Environments

    NASA Technical Reports Server (NTRS)

    Ladbury, R. L.; Lauenstein, J.-M.

    2016-01-01

    We develop metrics for assessing the effectiveness of proton SEE data for bounding heavy-ion SEE susceptibility. The metrics range from simple geometric criteria requiring no knowledge of the test articles to bounds of SEE rates.

  13. Tearing up the disc: misaligned accretion on to a binary

    NASA Astrophysics Data System (ADS)

    Nixon, Chris; King, Andrew; Price, Daniel

    2013-09-01

    In a recent paper, we have shown that the evolution of a misaligned disc around a spinning black hole can result in tearing the disc into many distinct planes. Tearing discs with random orientations produce direct dynamical accretion on to the hole in ≈70 per cent of all cases. Here, we examine the evolution of a misaligned disc around a binary system. We show that these discs are susceptible to tearing for almost all inclinations. We also show that tearing of the disc can result in a significant acceleration of the disc evolution and subsequent accretion on to the binary - by factors up to 104 times that of a coplanar prograde disc with otherwise identical parameters. This provides a promising mechanism for driving mergers of supermassive black hole (SMBH) binaries on time-scales much shorter than a Hubble time. Disc tearing also suggests new observational signatures of accreting SMBH binaries and other systems such as protostellar binaries.

  14. Physically-based landslide susceptibility modelling: geotechnical testing and model evaluation issues

    NASA Astrophysics Data System (ADS)

    Marchesini, Ivan; Mergili, Martin; Schneider-Muntau, Barbara; Alvioli, Massimiliano; Rossi, Mauro; Guzzetti, Fausto

    2015-04-01

    We used the software r.slope.stability for physically-based landslide susceptibility modelling in the 90 km² Collazzone area, Central Italy, exploiting a comprehensive set of lithological, geotechnical, and landslide inventory data. The model results were evaluated against the inventory. r.slope.stability is a GIS-supported tool for modelling shallow and deep-seated slope stability and slope failure probability at comparatively broad scales. Developed as a raster module of the GRASS GIS software, r.slope.stability evaluates the slope stability for a large number of randomly selected ellipsoidal potential sliding surfaces. The bottom of the soil (for shallow slope stability) or the bedding planes of lithological layers (for deep-seated slope stability) are taken as potential sliding surfaces by truncating the ellipsoids, allowing for the analysis of relatively complex geological structures. To take account for the uncertain geotechnical and geometric parameters, r.slope.stability computes the slope failure probability by testing multiple parameter combinations sampled deterministically or stochastically, and evaluating the ratio between the number of parameter combinations yielding a factor of safety below 1 and the total number of tested combinations. Any single raster cell may be intersected by multiple sliding surfaces, each associated with a slope failure probability. The most critical sliding surface is relevant for each pixel. Intensive use of r.slope.stability in the Collazzone Area has opened up two questions elaborated in the present work: (i) To what extent does a larger number of geotechnical tests help to better constrain the geotechnical characteristics of the study area and, consequently, to improve the model results? The ranges of values of cohesion and angle of internal friction obtained through 13 direct shear tests corresponds remarkably well to the range of values suggested by a geotechnical textbook. We elaborate how far an increased number of

  15. Antimicrobial Drug Susceptibility of Bacteria Isolated from Disease Processes in Cattle, Horses, Dogs and Cats

    PubMed Central

    Prescott, J. F.; Gannon, V. P.; Kittler, G.; Hlywka, G.

    1984-01-01

    Results are presented of antimicrobial disc diffusion susceptibility testing on commonly isolated bacterial pathogens made at the Ontario Veterinary College Diagnostic Bacteriology Laboratory in 1981 and 1982. Nearly 2 000 isolates from horses, cattle, dogs and cats were tested. Comparison of resistance in the same bacterial species isolated from different animal species showed significant differences between some of the same antibiotics. PMID:17422428

  16. ESBL Detection: Comparison of a Commercially Available Chromogenic Test for Third Generation Cephalosporine Resistance and Automated Susceptibility Testing in Enterobactericeae

    PubMed Central

    El-Jade, Mohamed Ramadan; Parcina, Marijo; Schmithausen, Ricarda Maria; Stein, Christoph; Meilaender, Alina; Hoerauf, Achim; Molitor, Ernst

    2016-01-01

    Rapid detection and reporting of third generation cephalosporine resistance (3GC-R) and of extended spectrum betalactamases in Enterobacteriaceae (ESBL-E) is a diagnostic and therapeutic priority to avoid inefficacy of the initial antibiotic regimen. In this study we evaluated a commercially available chromogenic screen for 3GC-R as a predictive and/or confirmatory test for ESBL and AmpC activity in clinical and veterinary Enterobacteriaceae isolates. The test was highly reliable in the prediction of cefotaxime and cefpodoxime resistance, but there was no correlation with ceftazidime and piperacillin/tazobactam minimal inhibitory concentrations. All human and porcine ESBL-E tested were detected with exception of one genetically positive but phenotypically negative isolate. By contrast, AmpC detection rates lay below 30%. Notably, exclusion of piperacillin/tazobactam resistant, 3GC susceptible K1+ Klebsiella isolates increased the sensitivity and specificity of the test for ESBL detection. Our data further imply that in regions with low prevalence of AmpC and K1 positive E. coli strains chromogenic testing for 3GC-R can substitute for more time consuming ESBL confirmative testing in E. coli isolates tested positive by Phoenix or VITEK2 ESBL screen. We, therefore, suggest a diagnostic algorithm that distinguishes 3GC-R screening from primary culture and species-dependent confirmatory ESBL testing by βLACTATM and discuss the implications of MIC distribution results on the choice of antibiotic regimen. PMID:27494134

  17. ESBL Detection: Comparison of a Commercially Available Chromogenic Test for Third Generation Cephalosporine Resistance and Automated Susceptibility Testing in Enterobactericeae.

    PubMed

    El-Jade, Mohamed Ramadan; Parcina, Marijo; Schmithausen, Ricarda Maria; Stein, Christoph; Meilaender, Alina; Hoerauf, Achim; Molitor, Ernst; Bekeredjian-Ding, Isabelle

    2016-01-01

    Rapid detection and reporting of third generation cephalosporine resistance (3GC-R) and of extended spectrum betalactamases in Enterobacteriaceae (ESBL-E) is a diagnostic and therapeutic priority to avoid inefficacy of the initial antibiotic regimen. In this study we evaluated a commercially available chromogenic screen for 3GC-R as a predictive and/or confirmatory test for ESBL and AmpC activity in clinical and veterinary Enterobacteriaceae isolates. The test was highly reliable in the prediction of cefotaxime and cefpodoxime resistance, but there was no correlation with ceftazidime and piperacillin/tazobactam minimal inhibitory concentrations. All human and porcine ESBL-E tested were detected with exception of one genetically positive but phenotypically negative isolate. By contrast, AmpC detection rates lay below 30%. Notably, exclusion of piperacillin/tazobactam resistant, 3GC susceptible K1+ Klebsiella isolates increased the sensitivity and specificity of the test for ESBL detection. Our data further imply that in regions with low prevalence of AmpC and K1 positive E. coli strains chromogenic testing for 3GC-R can substitute for more time consuming ESBL confirmative testing in E. coli isolates tested positive by Phoenix or VITEK2 ESBL screen. We, therefore, suggest a diagnostic algorithm that distinguishes 3GC-R screening from primary culture and species-dependent confirmatory ESBL testing by βLACTATM and discuss the implications of MIC distribution results on the choice of antibiotic regimen. PMID:27494134

  18. Susceptibility testing of Actinobacillus pleuropneumoniae in Denmark. Evaluation of three different media of MIC-determinations and tablet diffusion tests.

    PubMed

    Aarestrup, F M; Jensen, N E

    1999-02-12

    This study was conducted to compare the applicability of three different media in sensitivity testing of Actinobacillus pleuropneumoniae by means of MIC and tablet diffusion tests. The media used were: modified PPLO agar, chocolatized Mueller-Hinton-II and Columbia agar supplemented with NAD. Seven antimicrobial agents were tested: ceftiofur, enrofloxacin, penicillin, spectinomycin, tiamulin, trimethoprim + sulfadiazine and tylosin, against 40 randomly selected A. pleuropneumoniae isolates. In general, good agreement was found between results obtained with all combinations of media, most antimicrobials tested and the two-test systems. Some variations between media were observed for spectinomycin, tiamulin and tylosin. For ceftiofur and trimethoprim + sulfadiazine some isolates with low MIC-values were classified as resistant using tablet diffusion, indicating that the break points of resistance for these antimicrobials using the tablet diffusion tests need adjustment. Using current break points for resistance with MIC-determinations, all isolates tested susceptible to ceftiofur, enrofloxacin, penicillin, tiamulin and trimethoprim + sulfadiazine. A larger number of isolates tested resistant to spectinomycin and tylosin on all three media using both MIC determinations and tablet diffusion. PMID:10063535

  19. Comparison of Etest with Modified Broth Microdilution Method for Testing Susceptibility of Aspergillus spp. to Voriconazole

    PubMed Central

    Serrano, M. C.; Morilla, D.; Valverde, A.; Chávez, M.; Espinel-Ingroff, A.; Claro, R.; Ramírez, M.; Mazuelos, E. Martín

    2003-01-01

    We compared the Etest with a broth microdilution method, performed according to a modified National Committee for Clinical Laboratory Standards guideline (M38-A), for determining the in vitro susceptibility of 77 isolates of Aspergillus spp. (26 A. fumigatus, 21 A. flavus, 10 A. terreus, 9 A. niger, 5 A. nidulellus, 4 A. glaucus, and 2 A. flavipes isolates). Overall, there was 92.2% agreement between both methods when Etest MICs were read at 24 h and 83.1% agreement when both methods were read at 48 h. When Etest MICs were read at 24 h, the agreement was >90% for all species tested except for A. fumigatus (84.6%). When Etest MICs were read at 48 h, the agreement ranged from 50 to 100%. The poorest agreement was seen with A. glaucus (50%) and A. fumigatus (65%). Where a discrepancy was observed between Etest and the reference method, the Etest MIC was generally higher. The Etest appears to be a suitable alternative procedure for testing the susceptibility of Aspergillus spp. to voriconazole. PMID:14605181

  20. Rapid antimicrobial susceptibility testing of clinical isolates by digital time-lapse microscopy.

    PubMed

    Fredborg, M; Rosenvinge, F S; Spillum, E; Kroghsbo, S; Wang, M; Sondergaard, T E

    2015-12-01

    Rapid antimicrobial susceptibility testing (AST) is essential for early and appropriate therapy. Methods with short detection time enabling same-day treatment optimisation are highly favourable. In this study, we evaluated the potential of a digital time-lapse microscope system, the oCelloScope system, to perform rapid AST. The oCelloScope system demonstrated a very high accuracy (96% overall agreement) when determining the resistance profiles of four reference strains, nine clinical isolates, including multi-drug-resistant isolates, and three positive blood cultures. AST of clinical isolates (168 antimicrobial agent-organism combinations) demonstrated 3.6% minor, no major and 1.2% very major errors of the oCelloScope system compared to conventional susceptibility testing, as well as a rapid and correct phenotypic detection of strains with methicillin-resistant Staphylococcus aureus (MRSA) and extended-spectrum β-lactamase (ESBL) profiles. The net average time-to-result was 108 min, with 95% of the results being available within 180 min. In conclusion, this study strongly indicates that the oCelloScope system holds considerable potential as an accurate and sensitive AST method with short time-to-result, enabling same-day targeted antimicrobial therapy, facilitating antibiotic stewardship and better patient management. A full-scale validation of the oCelloScope system including more isolates is necessary to assess the impact of using it for AST. PMID:26407621

  1. Rapid purity check method for susceptibility testing of M. tuberculosis complex with the MGIT 960 system.

    PubMed

    Huang, Tsi-Shu; Liu, Yung-Ching; Tu, Hui-Zin; Sy, Cheng-Len; Chen, Yao-Shen; Chen, Bao-Chen

    2007-01-01

    The Bactec MGIT 960 system is a rapid and reliable automated method for drug susceptibility testing of Mycobacterium tuberculosis complex (MTBC) that yields a high percentage of agreement with the standard method. The microscopic cord morphology of M. tuberculosis in liquid medium is characteristic, and readily differentiates MTBC from nontuberculous mycobacteria (NTM). The goals of this study were to describe the microscopic and macroscopic growth morphology of MTBC in antimicrobial-containing MGIT tubes and to evaluate the usefulness of the growth appearance during purity checking. The macroscopic cotton wool-like appearance of MTBC isolates in isoniazid (INH), streptomycin (SM), rifampin (RMP), and ethambutol (EMB)-containing tubes was observed in 97, 90, 93, and 71% of the isolates, respectively. The percentage of typical cord, loose, or frayed rope microscopic features in smears prepared from MTBC-positive cultures of INH, SM, RMP, and EMB-containing tubes was 96, 86, 97, and 71%, respectively. The sensitivity of the macroscopic morphology for predicting the purity of drug-containing MGIT tubes was 93%, while the microscopic morphology predicted the purity with a sensitivity rate of 92%. We found that simply examining the macroscopic morphology of the antimicrobial-containing MGIT tubes of drug-resistant MTBC isolates is useful in preventing false resistant results of susceptibility testing by the MGIT 960 system. PMID:18000288

  2. Janus discs.

    PubMed

    Walther, Andreas; André, Xavier; Drechsler, Markus; Abetz, Volker; Müller, Axel H E

    2007-05-16

    We describe the synthesis and the solution properties of sheet- and disclike Janus particles, containing an inner crosslinked polybutadiene (PB) layer and two different outer sides of polystyrene (PS) and poly(tert-butyl methacrylate) (PtBMA). The structures formed upon adsorption of the flat Janus particles onto solid substrates as well as in THF solution are investigated. The Janus discs are obtained in a template-assisted synthetic pathway followed by sonication. Selectively crosslinking the lamellar PB domains in a well-ordered lamellar microphase-separated bulk morphology of PS-block-PB-block-PtBMA (SBT) block terpolymers leads to the conservation of the compartmentalization of the two outer blocks. Sonication of the crosslinked block terpolymer templates renders soluble sheet- and disclike Janus particles, the size of which can be tuned from the micrometer range down to the nanometer scale. Small-angle X-ray scattering, transmission electron microscopy, dynamic light scattering, scanning force microscopy, and scanning electron microscopy are used to characterize the template-assisted synthetic process and the solution properties. Cryogenic transmission electron microscopy in THF and TEM of particles, embedded into a photo-crosslinkable silicon oil, indicate a supramolecular aggregation behavior of the Janus discs in concentrated solutions. Pendant drop tensiometry demonstrates that Janus sheets and discs can be used to stabilize liquid-liquid interfaces, rendering these materials interesting for future applications. PMID:17441717

  3. Coagulase-negative staphylococci from non-mastitic bovine mammary gland: characterization of Staphylococcus chromogenes and Staphylococcus haemolyticus by antibiotic susceptibility testing and pulsed-field gel electrophoresis.

    PubMed

    Pate, Mateja; Zdovc, Irena; Avberšek, Jana; Ocepek, Matjaž; Pengov, Andrej; Podpečan, Ožbalt

    2012-05-01

    During routine microbiological examination of milk samples from dairy cows without clinical signs of mastitis, quarter milk samples of 231 dairy cows from 12 herds were investigated for the presence of coagulase-negative staphylococci (CNS). The isolates were identified on the basis of colony morphology, Gram staining, catalase and coagulase test and the commercial kit, API Staph. CNS was detected in 29% (67/231) of the cows. A total of seven CNS species were identified with the most prevalent being Staphylococcus (Staph.) chromogenes (30%) and Staph. haemolyticus (28·8%), followed by Staph. simulans (11·2%), Staph. xylosus (11·2%), Staph. epidermidis (7·5%), Staph. hyicus (6·3%) and Staph. sciuri (5%). The predominant species, Staph. chromogenes and Staph. haemolyticus, were further characterized by antibiotic susceptibility testing using the agar disc diffusion method (Kirby-Bauer) and by pulsed-field gel electrophoresis (PFGE). Considerable resistance to ampicillin and penicillin was observed in both species. Isolates with identical or highly similar PFGE profiles were detected at the herd level despite a marked heterogeneity seen for both species. On the basis of somatic cell count, absence of clinical signs of inflammation and heterogeneity of genotypes, we assume that CNS isolated in this study could not be considered as important causative agents of the bovine mammary gland inflammation. PMID:22067091

  4. Development of an Antimicrobial Susceptibility Testing Method Suitable for Performing During Space Flight

    NASA Technical Reports Server (NTRS)

    Jorgensen, James H.; Skweres, Joyce A.; Mishra S. K.; McElmeel, M. Letticia; Maher, Louise A.; Mulder, Ross; Lancaster, Michael V.; Pierson, Duane L.

    1997-01-01

    Very little is known regarding the affects of the microgravity environment of space flight upon the action of antimicrobial agents on bacterial pathogens. This study was undertaken to develop a simple method for conducting antibacterial susceptibility tests during a Space Shuttle mission. Specially prepared susceptibility test research cards (bioMerieux Vitek, Hazelwood, MO) were designed to include 6-11 serial two-fold dilutions of 14 antimicrobial agents, including penicillins, cephalosporins, a Beta-lactamase inhibitor, vancomycin, erythromycin, tetracycline, gentamicin, ciprofloxacin, and trimethoprim/sulfamethoxazole. Minimal inhibitory concentrations (MICS) of the drugs were determined by visual reading of color endpoints in the Vitek research cards made possible by incorporation of a colorimetric growth indicator (alamarBlue(Trademark), Accumed International, Westlake, OH). This study has demonstrated reproducible susceptibility results when testing isolates of Staphylococcus aurezis, Group A Streptococcus, Enterococcusfaecalis, Escherichia coli (beta-lactamase positive and negative strains), Klebsiella pneumoniae, Enterobacter cloacae, and Pseudomoiias aeruginosa. In some instances, the MICs were comparable to those determined using a standard broth microdilution method, while in some cases the unique test media and format yielded slightly different values, that were themselves reproducible. The proposed in-flight experiment will include inoculation of the Vitek cards on the ground prior to launch of the Space Shuttle, storage of inoculated cards at refrigeration temperature aboard the Space Shuttle until experiment initiation, then incubation of the cards for 18-48 h prior to visual interpretation of MICs by the mission's astronauts. Ground-based studies have shown reproducible MICs following storage of inoculated cards for 7 days at 4-8 C to accommodate the mission's time schedule and the astronauts' activities. For comparison, ground-based control

  5. Methicillin Resistant Staphylococcus aureus: Inconsistencies in Vancomycin Susceptibility Testing Methods, Limitations and Advantages of each Method

    PubMed Central

    Himani; Madan, Molly; Pandey, Anita; Thakuria, Bhaskar

    2015-01-01

    Background Vancomycin may be ineffective against an increasing proportion of methicillin resistant Staphylococcus aureus (MRSA) with minimum inhibitory concentrations (MICs) well within the susceptible range. On the other hand it is common knowledge that determination of vancomycin MICs is method dependent. Therefore, given the apparent variability in vancomycin MIC results obtained with the different methods, the use of the vancomycin MIC to predict the outcome of serious S. aureus infections needs to take into account the method used and the results of studies using that particular method. Aim Comparative study was carried out to evaluate the MICs obtained by BMD method, E-test, and Vitek 2 method and to detect inconsistencies in these vancomycin for 66 MRSA isolates obtained from various samples of patients attending the OPDs & IPDs within a period of one year. Materials and Methods A comparative study was carried out to evaluate the MICs obtained by BMD method, E-test, and Vitek 2 method to detect vancomycin susceptibility in 66 clinical isolates of MRSA obtained from various samples of patients attending the OPDs & IPDs within a period of one year. The study was conducted in Department of Microbiology, Subharti Medical College, Meerut from January to December 2012. Results On determination of MICs for vancomycin for the MRSA isolates, all were identified as VSSA by BMD, E-Test & Vitek 2 methods. However, the vancomycin MIC values obtained by E-test correlated better with BMD method (correlation factor= 0.6727) than Vitek 2 (correlation factor=0.5316), indicating E-Test to be a better method for determination of vancomycin MICs as compared to Vitek 2. Conclusion MRSA isolates with higher vancomycin MICs, even within the susceptibility range, are being observed more frequently which result in treatment failures with vancomycin. Because of the discrepancy that exists in vancomycin MIC results from different methods, the prediction of outcome of serious S

  6. Reliability of Disk Diffusion Test Results for the Antimicrobial Susceptibility Testing of Nosocomial Gram-positive Microorganisms: Is E-test Method Better?

    PubMed Central

    Khalili, Hossein; Soltani, Rasool; Negahban, Sorrosh; Abdollahi, Alireza; Gholami, Keirollah

    2012-01-01

    Disk diffusion test is the usual applicable method for assessing the antimicrobial susceptibility pattern in most institutions and hospitals. The aim of this study was to determine the reliability of resistant-reported results of disk diffusion test for 6 routinely used antibiotics against Gram-positive microorganisms of nosocomial origin, using E-test method. Over a 1-year period, clinical specimens (e.g. blood, tracheal secretions, wound secretions, urine, etc.) were obtained from hospitalized patients with defined nosocomial infection and were cultured. Isolated Gram-positive bacteria underwent disk diffusion test for cephalothin, oxacillin, clindamycin, ciprofloxacin, vancomycin, teicoplanin (only for Enterococci), and meropenem antibiotics. E-test method was performed for all isolates resistant or intermediately sensitive to the disks of any mentioned antibiotics. Data showed compatible results of disk diffusion test with the results of E-test method for cephalothin, oxacillin, ciprofloxacin, vancomycin, and teicoplanin. None of ciprofloxacin- and vancomycin-resistant isolates in disk diffusion test showed sensitivity in E-test method. Significant differences between the results of disk diffusion and E-test methods were observed for clindamycin and meropenem against S.aureus (p = 0.01 and 0.04, respectively) and Enterococcus spp (p = 0.03 and 0.02, respectively). In order to increase the reliability of antimicrobial susceptibility results, it is recommended to perform E-test for nosocomial Gram-positive microorganisms that show antibiotic resistance by disk diffusion test and it is more important for clindamycin and meropenem. PMID:24250479

  7. Susceptibility testing of filamentous fungi to amphotericin B by a rapid radiometric method

    SciTech Connect

    Merz, W.G.; Fay, D.; Thumar, B.; Dixon, D.

    1984-01-01

    A rapid, radiometric method was developed to determine the susceptibility of filamentous fungi to amphotericin B. The rapid, radiometric method depended on measurement of the inhibition of /sup 24/CO/sub 2/ production in the presence of amphotericin B. Thirty isolates of filamentous fungi were tested by the rapid, radiometric method and a reference agar dilution method. There was 93% agreement between the two methods when an 80% or greater decrease in CO/sub 2/ production was used to calculate the minimal inhibitory concentration with the rapid, radiometric method. Minimal inhibitory concentrations, based on 80% decrease of CO/sub 2/ production, were achieved within 24 h of incubation with all of the fungi tested.

  8. Comparison of E-test with broth microdilution and disk diffusion for susceptibility testing of coryneform bacteria.

    PubMed

    Martínez-Martínez, L; Ortega, M C; Suárez, A I

    1995-05-01

    The susceptibilities of 135 coryneform bacteria isolated from clinical samples to ampicillin (AMP), cephalothin (CR), cefoxitin (FOX), cefotaxime (CTX), erythromycin (E), ciprofloxacin (CIP), tetracycline (TE), amikacin (AK), vancomycin (VA), and rifampin (R) were determined by disk diffusion, broth microdilution, and the E-test. The following species (number of isolates in parentheses) were included: Corynebacterium urealyticum (30), Corynebacterium minutissimum (20), coryneform CDC group ANF-1 (20), Corynebacterium striatum (20), Corynebacterium jeikeium (15), coryneform CDC group I2 (8), Listeria monocytogenes (7), Corynebacterium xerosis (5), and other coryneform bacteria (10). Agreement within one twofold dilution between the E-test and broth microdilution was 31% (VA), 64% (AK), 71% (CTX), 77% (FOX and CIP), 79% (TE), 84% (AMP), 87% (E), and 88% (CR and R). For the 1,350 combinations of microorganisms and antimicrobial agents, 85 (6.3%) discrepancies in interpretive category were found (4.2% minor, 1.2% major, and 0.9% very major). Seventy (5.1%) disagreements in interpretive category were found between disk diffusion and the E-test (3.8% minor, 0.4% major, and 0.9% very major), and 85 (6.3%) disagreements were found between microdilution (reference method) and disk diffusion (4.2% minor, 0.5% major, and 1.5% very major). MICs obtained with the E-test were highly reproducible. No category discrepancy was observed for VA, despite quantitative results. Considering interpretive categories, there is a good overall agreement between the three methods studied here, but further evaluation of current methodologies for susceptibility testing is required when considering coryneform bacteria and determination of quantitative activity of antimicrobial agents. PMID:7615748

  9. Performance of tuberculosis drug susceptibility testing in U.S. laboratories from 1994 to 2008.

    PubMed

    Angra, Pawan K; Taylor, Thomas H; Iademarco, Michael F; Metchock, Beverly; Astles, J Rex; Ridderhof, John C

    2012-04-01

    We present a statistical summary of results from the Model Performance Evaluation Program (MPEP) for Mycobacterium tuberculosis Drug Susceptibility Testing, 1994 to 2008, implemented by the U.S. Centers for Disease Control and Prevention (CDC). During that period, a total of 57,733 test results for culture isolates were reported by 216 participating laboratories for the first-line antituberculosis drugs used in the United States-isoniazid (INH), rifampin (RMP), ethambutol (EMB), and pyrazinamide (PZA). Using Clinical Laboratory and Standards Institute (CLSI)-recommended concentrations for one or more of three methods, agar proportion (AP), BACTEC460 (Bactec), and MGIT-960 (MGIT), yielded overall agreement of 97.0% for first-line drugs. For susceptible strains, agreement was 98.4%; for resistant strains, agreement was 91.0%, with significantly lower accuracy (chi-square test, P < 0.0001). For resistant strains, overall agreement by methods was 91.3% for AP, 93.0% for Bactec, and 82.6% for MGIT and by drugs was 92.2% for INH, 91.5% for RMP, 79.0% for EMB, and 97.5% for PZA. For some strains, performance by method varied significantly. Use of duplicate strains in the same shipment and repeat strains over time revealed consistent performance even for strains with higher levels of interlaboratory discordance. No overall differences in performance between laboratories were observed based on volume of testing or type of facility (e.g., health department, hospital, independent). By all methods, decreased performance was observed for strains with low-level INH resistance, RMP resistance, and EMB-resistant strains. These results demonstrate a high level of performance in detection of drug-resistant M. tuberculosis in U.S. laboratories. PMID:22301024

  10. Target Product Profile of a Molecular Drug-Susceptibility Test for Use in Microscopy Centers

    PubMed Central

    Denkinger, Claudia M.; Dolinger, David; Schito, Marco; Wells, William; Cobelens, Frank; Pai, Madhukar; Zignol, Matteo; Cirillo, Daniela Maria; Alland, David; Casenghi, Martina; Gallarda, Jim; Boehme, Catharina C.; Perkins, Mark D.

    2015-01-01

    Background. Current phenotypic testing for drug resistance in patients with tuberculosis is inadequate primarily with respect to turnaround time. Molecular tests hold the promise of an improved time to diagnosis. Methods. A target product profile for a molecular drug-susceptibility test (DST) was developed on the basis of a collaborative effort that included opinions gathered from researchers, clinicians, policy makers, and test developers on optimal clinical and operational characteristics in settings of intended use. In addition, the current diagnostic ecosystem and the diagnostic development landscape were mapped. Results. Molecular DSTs for detecting tuberculosis in microscopy centers should ideally evaluate for resistance to rifampin, fluoroquinolones, isoniazid, and pyrazinamide and enable the selection of the most appropriate treatment regimen. Performance characteristics of DSTs need to be optimized, but compromises can be made that depend on the trade-off between a false-positive result and a false-negative result. The operational requirements of a test will vary depending on the site of implementation. However, the most-important considerations pertain to quality control, maintenance and calibration, and the ability to export data. Conclusion. This target product profile defines the needs as perceived by the tuberculosis stakeholder community and attempts to provide a means of communication with test developers to ensure that fit-for-purpose DSTs are being developed. PMID:25765105

  11. CHROMagar Candida Medium for Direct Susceptibility Testing of Yeast from Blood Cultures

    PubMed Central

    Tan, Grace L.; Peterson, Ellena M.

    2005-01-01

    An evaluation was performed on 95 blood cultures positive for Candida spp. to determine the correlation of direct susceptibility testing of fluconazole versus both standardized disk diffusion and MIC methods. For direct testing, an aliquot taken from BD BACTEC Plus and/or BD BACTEC Lytic/10 bottles (Becton Dickinson [BD], Sparks, MD) positive by gram stain for yeast was subcultured to CHROMagar Candida (BD), and a 25-μg fluconazole disk (BD) was placed on the plate. The area of growth inhibition surrounding the disk was measured at 24 and 48 h. In addition, a subculture of the isolate was tested by a microdilution MIC using YeastOne (TREK Diagnostics Systems Inc., OH) and disk diffusion (NCCLS M44-A) using a standardized inoculum plated onto CHROMagar Candida as well as Mueller-Hinton agar to which 2% glucose and 0.5 μg/ml methylene blue dye was added (MH-GMB). The categorical interpretation derived from the MIC was used as the reference to which the disk diffusion results were compared. There were a total of 41 Candida albicans, 23 Candida glabrata, 20 Candida parapsilosis, 9 Candida tropicalis, and 1 each of Candida krusei and Candida lusitaniae tested. At 24 h there was full agreement among the methods for all C. albicans, C. tropicalis, C. lusitaniae, and C. krusei isolates. For the C. parapsilosis isolates at 24 h there was one very major discrepancy using the direct CHROMagar and one major error with the standardized MH-GMB. The majority of the errors were seen at 24 h with the C. glabrata isolates. Of the 23 C. glabrata isolates at 24 h by direct CHROMagar, there were 10 minor and 1 very major error; by MH-GMB there were 12 minor and 2 very major errors; and by standardized CHROMagar Candida there were 13 minor and 2 major errors. There were no very major errors with C. glabrata when all plates were read at 48 h. At 24 h by the direct and standardized CHROMagar the majority of C. glabrata isolates were more resistant, whereas by MH-GMB they were more

  12. Radiorespirometric testing of antibiotic sensitivity in urinary tract infections: concise communication

    SciTech Connect

    Singh, K.T.; Ganatra, R.D.; Shah, D.H.; Shanta, M.; Nimbkar, Y.S.; Gaitonde, B.B.; Dudani, R.A.; Jadav, S.K.; Acharya, V.N.

    1980-02-01

    A radiometric method, based on inhibition of /sup 14/CO/sub 2/ release from bacterial metabolism of C-14-labeled glucose, was applied to test the susceptibility of urinary organisms to antibiotics. The testing was also carried out by the routine disc diffusion method after isolation of the organisms. Results of susceptibility to antibiotics could be obtained within 2 to 4 hr by the radiometric technique, compared with the 48 hr required for the disc method.

  13. Elucidating the relationship between DISC1, NDEL1 and NDE1 and the risk for schizophrenia: Evidence of epistasis and competitive binding

    PubMed Central

    Burdick, Katherine E.; Kamiya, Atsushi; Hodgkinson, Colin A.; Lencz, Todd; DeRosse, Pamela; Ishizuka, Koko; Elashvili, Sarah; Arai, Hiroyuki; Goldman, David; Sawa, Akira; Malhotra, Anil K.

    2008-01-01

    DISC1 influences susceptibility to psychiatric disease and related phenotypes. Intact functions of DISC1 and its binding partners, NDEL1 and NDE1, are critical to neurodevelopmental processes aberrant in schizophrenia (SZ). Despite evidence of an NDEL1–DISC1 protein interaction, there have been no investigations of the NDEL1 gene or the relationship between NDEL1 and DISC1 in SZ. We genotyped six NDEL1 single-nucleotide polymorphisms (SNPs) in 275 Caucasian SZ patients and 200 controls and tested for association and interaction between the functional SNP Ser704Cys in DISC1 and NDEL1. We also evaluated the relationship between NDE1 and DISC1 genotype and SZ. Finally, in a series of in vitro assays, we determined the binding profiles of NDEL1 and NDE1, in relation to DISC1 Ser704Cys. We observed a single haplotype block within NDEL1; the majority of variation was captured by NDEL1 rs1391768. We observed a significant interaction between rs1391768 and DISC1 Ser704Cys, with the effect of NDEL1 on SZ evident only against the background of DISC1 Ser704 homozygosity. Secondary analyses revealed no direct relationship between NDE1 genotype and SZ; however, there was an opposite pattern of risk for NDE1 genotype when conditioned on DISC1 Ser704Cys, with NDE1 rs3784859 imparting a significant effect but only in the context of a Cys-carrying background. In addition, we report opposing binding patterns of NDEL1 and NDE1 to Ser704 versus Cys704, at the same DISC1 binding domain. These data suggest that NDEL1 significantly influences risk for SZ via an interaction with DISC1. We propose a model where NDEL1 and NDE1 compete for binding with DISC1. PMID:18469341

  14. Genetic diversity and antifungal susceptibility testing of Trichosporon asahii isolated of Intensive Care Units patients

    PubMed Central

    de Oliveira Silva, Rosana Bellan; Fusco-Almeida, Ana Marisa; Matsumoto, Marcelo Teruyuki; Baeza, Lilian Cristiane; Benaducci, Tatiane; Mendes-Giannini, Maria José Soares

    2008-01-01

    Trichosporon asahii is an opportunistic pathogen, associated with a high mortality rate in immunocompromised patients. In this study, ten isolates, recovered from oral cavity and urine of patients in Intensive Care Units (ICU) over six months, were identified by classical and molecular methods, typed by RAPD and tested in vitro for susceptibility to fluconazole, itraconazole, 5-flucytosine and amphotericin B. A total agreement between the identification of Trichosporon sp by PCR based on sequences of the Internal Transcribed Spacer Regions (ITS) and on the sequences of small-subunit (SSU) ribosomal DNA (rDNA) was found. Randomly amplified of polymorphic DNA (RAPD), with primers P6 and M13, was used to determine the genomic profiles. The dendogram analysis indicated that almost all strains showed similarity >0.9 among them and all strains were multidrug-resistant. This study brings new results on the identification and genotyping of T. asahii isolated from Brazilian ICU patients and information about their antifungal drugs susceptibility. PMID:24031270

  15. Drug Susceptibility Testing of 31 Antimicrobial Agents on Rapidly Growing Mycobacteria Isolates from China

    PubMed Central

    Pang, Hui; Li, Guilian; Zhao, Xiuqin; Liu, Haican; Wan, Kanglin; Yu, Ping

    2015-01-01

    Objectives. Several species of rapidly growing mycobacteria (RGM) are now recognized as human pathogens. However, limited data on effective drug treatments against these organisms exists. Here, we describe the species distribution and drug susceptibility profiles of RGM clinical isolates collected from four southern Chinese provinces from January 2005 to December 2012. Methods. Clinical isolates (73) were subjected to in vitro testing with 31 antimicrobial agents using the cation-adjusted Mueller-Hinton broth microdilution method. The isolates included 55 M. abscessus, 11 M. fortuitum, 3 M. chelonae, 2 M. neoaurum, and 2 M. septicum isolates. Results. M. abscessus (75.34%) and M. fortuitum (15.07%), the most common species, exhibited greater antibiotic resistance than the other three species. The isolates had low resistance to amikacin, linezolid, and tigecycline, and high resistance to first-line antituberculous agents, amoxicillin-clavulanic acid, rifapentine, dapsone, thioacetazone, and pasiniazid. M. abscessus and M. fortuitum were highly resistant to ofloxacin and rifabutin, respectively. The isolates showed moderate resistance to the other antimicrobial agents. Conclusions. Our results suggest that tigecycline, linezolid, clofazimine, and cefmetazole are appropriate choices for M. abscessus infections. Capreomycin, sulfamethoxazole, tigecycline, clofazimine, and cefmetazole are potentially good choices for M. fortuitum infections. Our drug susceptibility data should be useful to clinicians. PMID:26351633

  16. Child exposure to serious life events, COMT, and aggression: Testing differential susceptibility theory.

    PubMed

    Hygen, Beate Wold; Belsky, Jay; Stenseng, Frode; Lydersen, Stian; Guzey, Ismail Cuneyt; Wichstrøm, Lars

    2015-08-01

    Both genetic and environmental factors contribute to individual differences in aggression. Catechol-O-methyltransferase Val158Met (COMT), a common, functional polymorphism, has been implicated in aggression and aggression traits, as have childhood experiences of adversity. It is unknown whether these effects are additive or interactional and, in the case of interaction, whether they conform to a diathesis-stress or differential susceptibility model. We examined Gene × Environment interactions between COMT and serious life events on measures of childhood aggression and contrasted these 2 models. The sample was composed of community children (N = 704); 355 were boys, and the mean age was 54.8 months (SD = 3.0). The children were genotyped for COMT rs4680 and assessed for serious life events and by teacher-rated aggression. Regression analysis showed no main effects of COMT and serious life events on aggression. However, a significant interactive effect of childhood serious life events and COMT genotype was observed: Children who had faced many serious life events and were Val homozygotes exhibited more aggression (p = .02) than did their Met-carrying counterparts. Notably, in the absence of serious life events, Val homozygotes displayed significantly lower aggression scores than did Met carriers (p = .03). When tested, this constellation of findings conformed to the differential susceptibility hypothesis: In this case, Val homozygotes are more malleable to the effect of serious life events on aggression and not simply more vulnerable to the negative effect of having experienced many serious life events. PMID:26053146

  17. Interpretive Breakpoints for Fluconazole and Candida Revisited: a Blueprint for the Future of Antifungal Susceptibility Testing

    PubMed Central

    Pfaller, M. A.; Diekema, D. J.; Sheehan, D. J.

    2006-01-01

    Developing interpretive breakpoints for any given organism-drug combination requires integration of the MIC distribution, pharmacokinetic and pharmacodynamic parameters, and the relationship between in vitro activity and outcome from both in vivo and clinical studies. Previously, the Subcommittee for Antifungal Testing of the Clinical and Laboratory Standards Institute (CLSI [formerly National Committee for Clinical Laboratory Standards]) proposed MIC interpretive breakpoints for fluconazole and Candida spp. These breakpoints were considered to be somewhat weak, because the clinical data supporting them came largely from mucosal infections and there were very few infections involving strains with elevated fluconazole MICs. We readdress the issue of fluconazole breakpoints for Candida by using published clinical and microbiologic data to provide further validation of the breakpoints proposed by the CLSI in 1997. We also address interpretive breakpoints for agar disk diffusion testing of fluconazole. The MIC distribution for fluconazole was determined with a collection of 13,338 clinical isolates. The overall MIC at which 90% of the isolates were inhibited was 8 μg/ml: 91% were susceptible (S) at a MIC of ≤8 μg/ml and 3% were resistant (R) (MIC ≥ 64 μg/ml). Similar results were obtained for 2,190 isolates from randomized clinical trials. Analysis of available data for 1,295 patient-episode-isolate events (692 represented mucosal infections and 603 represented invasive infections) from 12 published clinical studies demonstrated an overall success rate of 77%, including 85% for those episodes in which the fluconazole MIC was ≤8 μg/ml, 67% for those episodes in which the MIC was 16 to 32 μg/ml, and 42% for those episodes with resistant (MIC ≥ 64 μg/ml) isolates. Pharmacodynamic analysis demonstrated a strong relationship between MIC, fluconazole dose, and outcome. A dose/MIC ratio of ∼25 was supportive of the following susceptibility breakpoints for

  18. Towards in vitro DT/DNT testing: Assaying chemical susceptibility in early differentiating NT2 cells.

    PubMed

    Menzner, Ann-Katrin; Abolpour Mofrad, Sepideh; Friedrich, Oliver; Gilbert, Daniel F

    2015-12-01

    Human pluripotent embryonal carcinoma (NT2) cells are increasingly considered as a suitable model for in vitro toxicity testing, e.g. developmental toxicity and neurotoxicity (DT/DNT) studies, as they undergo neuronal differentiation upon stimulation with retinoic acid (RA) and permit toxicity testing at different stages of maturation. NT2 cells have recently been reported to show specific changes in dielectric resistance profiles during differentiation which can be observed as early as 24h upon RA-stimulation. These observations suggest altered susceptibility to chemicals at an early stage of differentiation. However, chemical susceptibility of early differentiating NT cells has not yet been studied. To address this question, we have established a cell fitness screening assay based on the analysis of intracellular ATP levels and we applied the assay in a large-scale drug screening experiment in NT2 stem cells and early differentiating NT2 cells. Subsequent analysis of ranked fitness phenotypes revealed 19 chemicals with differential toxicity profile in early differentiating NT2 cells. To evaluate whether any of the identified drugs have previously been associated with DT/DNT, we conducted a literature search on the identified molecules and quantified the fraction of chemicals assigned to the FDA (Food and Drug Administration) pregnancy risk categories (PRC) N, A, B, C, D, and X in the hit list and the small molecule library. While the fractions of the categories N and B were decreased (0.81 and 0.35-fold), the classes C, D and X were increased (1.35, 1.47 and 3.27-fold) in the hit list compared to the chemical library. From these data as well as from the literature review, identifying large fractions of chemicals being directly (∼42%) and indirectly associated with DT/DNT (∼32%), we conclude that our method may be beneficial to systematic in vitro-based primary screening for developmental toxicants and neurotoxicants and we propose cell fitness screening in

  19. Multicenter evaluation of a broth macrodilution antifungal susceptibility test for yeasts.

    PubMed Central

    Fromtling, R A; Galgiani, J N; Pfaller, M A; Espinel-Ingroff, A; Bartizal, K F; Bartlett, M S; Body, B A; Frey, C; Hall, G; Roberts, G D

    1993-01-01

    Thirteen laboratories collaborated to optimize interlaboratory agreement of results of a broth macrodilution procedure for testing three classes of antifungal drugs against pathogenic yeasts. The activities of amphotericin B, flucytosine, and ketoconazole were tested against 100 coded isolates of Candida albicans, Candida tropicalis, Candida parapsilosis, Candida lusitaniae, Torulopsis (Candida) glabrata, and Cryptococcus neoformans. Two starting yeast inoculum sizes (5 x 10(4) and 2.5 x 10(3) cells per ml) were compared, and readings were taken after 24 and 48 h of incubation. All other test conditions were standardized. The resultant turbidities in all tubes were estimated visually on a scale from 0 to 4+ turbidity, and MIC-0, MIC-1, and MIC-2 were defined as the lowest drug concentrations that reduced growth to 0, 1+, or 2+ turbidity, respectively. For flucytosine, agreement among laboratories varied between 57 and 87% for different inocula, times of incubation, and end point criteria. Agreement was maximized (85%) when the lower inoculum was incubated for 2 days and the MICs were defined as 1+ turbidity or less. For amphotericin B, variations in test conditions produced much smaller differences in interlaboratory agreement. For ketoconazole, interlaboratory agreement was poorer by all end point criteria. However, MIC-2 endpoints distinguished T. glabrata as resistant compared with the other species. Overall, the studies indicated that readings from the lower inoculum obtained on the second day of reading result in the greatest interlaboratory agreement. In combination with data from previous multicenter studies (National Committee for Clinical Laboratory Standards, Antifungal Susceptibility Testing: Committee Report, Vol. 5, No. 17, 1988; M. A. Pfaller, L. Burmeister, M. S. Bartlett, and M. G. Rinaldi, J. Clin. Microbiol. 26:1437-1441, 1988; M. A. Pfaller, M. G. Rinaldi, J. N. Galgiani, M. S. Bartlett, B.A. Body, A. Espinel-Ingroff, R.A. Fromtling, G.S. Hall, C

  20. Miniaturized Antimicrobial Susceptibility Test by Combining Concentration Gradient Generation and Rapid Cell Culturing

    PubMed Central

    Kim, Samuel C.; Cestellos-Blanco, Stefano; Inoue, Keisuke; Zare, Richard N.

    2015-01-01

    Effective treatment of bacterial infection relies on timely diagnosis and proper prescription of antibiotic drugs. The antimicrobial susceptibility test (AST) is one of the most crucial experimental procedures, providing the baseline information for choosing effective antibiotic agents and their dosages. Conventional methods, however, require long incubation times or significant instrumentation costs to obtain test results. We propose a lab-on-a-chip approach to perform AST in a simple, economic, and rapid manner. Our assay platform miniaturizes the standard broth microdilution method on a microfluidic device (20 × 20 mm) that generates an antibiotic concentration gradient and delivers antibiotic-containing culture media to eight 30-nL chambers for cell culture. When tested with 20 μL samples of a model bacterial strain (E. coli ATCC 25922) treated with ampicillin or streptomycin, our method allows for the determination of minimum inhibitory concentrations consistent with the microdilution test in three hours, which is almost a factor of ten more rapid than the standard method. PMID:27025635

  1. Oral colonization and susceptibility testing of Pseudomonas aeruginosa oral isolates from cystic fibrosis patients.

    PubMed

    Lindemann, R A; Newman, M G; Kaufman, A K; Le, T V

    1985-01-01

    Microbial samples from the oral cavities of cystic fibrosis (C.F.) patients and 20 age-matched normal control subjects were characterized. Mucoid variant Pseudomonas aeruginosa was isolated from the tongue, buccal mucosa, and saliva of C.F. patients only. Analysis of the data suggests that the oral cavity is a potential reservoir for this organism. Aspiration and cross-contamination from this reservoir may be important in perpetuating chronic pulmonary infection in C.F. patients. Susceptibility testing was performed on 20 mucoid variant P. aeruginosa oral isolates obtained from the patients according to standardized broth dilution procedures. The in vitro antimicrobial effects of sodium fluoride, stannous fluoride, and chlorhexidine were measured. Analysis of the data suggests that clinically safe and achievable levels of chlorhexidine and stannous fluoride may be antimicrobial. PMID:3918088

  2. MBT-ASTRA: A suitable tool for fast antibiotic susceptibility testing?

    PubMed

    Sparbier, Katrin; Schubert, Sören; Kostrzewa, Markus

    2016-07-15

    The increasing resistance to antibiotics is an urgent health care problem. Detection of resistant microorganisms is the pre-requisite for initiating an adequate therapy and implementing respective hygiene measures. Depending on the species and the method employed for analysis, the time to result of antibiotic resistance testing ranges between five and 24h. As MALDI-TOF MS has become an established tool for the fast species identification in microbiological laboratories a time gap between the results of species identification and the information about antibiotic susceptibility arises. Here, we present a semi-quantitative MALDI-TOF MS-based approach for the detection of resistance in different species against different antibiotics. PMID:26804565

  3. In vitro antifungal susceptibility testing of Scopulariopsis brevicaulis strains using agar diffusion method.

    PubMed

    Skóra, Magdalena; Macura, Anna B

    2011-01-01

    The genus Scopulariopsis is a common soil saprotroph and has been isolated from air, organic waste and also from plant, animal and human tissues. Scopulariopsis has mainly been associated in humans with superficial mycoses, but it has also been described as the cause of subcutaneous and invasive infections. The most common aetiological agent of infections in humans is Scopulariopsis brevicaulis. This species has been reported to be resistant in vitro to broad-spectrum antifungal agents available today. The aim of the study was to establish in vitro antifungal susceptibility of 35 S. brevicaulis strains against amphotericin B (AMB), flucytosine (FC), caspofungin (CAS), terbinafine (TER), ciclopirox (CIC), voriconazole (VOR), clotrimazole (CTR), miconazole (MCZ), econazole (ECO), ketoconazole (KET), itraconazole (ITR), and fluconazole (FLU). Antifungal susceptibility tests were evaluated by an agar diffusion method (Neo-Sensitabs, Rosco, Denmark). AMB, FC, CAS, ITR and FLU showed no antifungal activity against S. brevicaulis. TER, CIC, CTR, KET, VOR, ECO, and MCZ revealed inhibitory activity for S. brevicaulis, but it varied for each of the drugs. The best antifungal effect was observed for TER and CIC. All isolates had large inhibition zones for TER and CIC. CTR was also inhibitory for all tested S. brevicaulis isolates, but the diameters of inhibition zones were smaller than for TER and CIC. Nearly 89% isolates showed inhibition zones for KET and the mean diameter of the inhibition zone was comparable to CTR. The least antifungal activity exhibited VQR, ECO and MCZ. Because of the multiresistance of S. brevicaulis, infections due to this species may not respond to particular antifungal treatment and other therapeutic approaches should be considered, e.g., combined therapy and/or surgery. PMID:21682097

  4. Artificial Disc Replacement

    MedlinePlus

    ... treat this condition, alternatives to disc replacement include fusion, nonoperative care or no treatment. Typically, surgery is ... operative treatment for disc pain has been spinal fusion. This is a surgical procedure in which disc ...

  5. Agreement of Direct Antifungal Susceptibility Testing from Positive Blood Culture Bottles with the Conventional Method for Candida Species.

    PubMed

    Jabeen, Kauser; Kumar, Haresh; Farooqi, Joveria; Mehboob, Raunaq; Brandt, Mary E; Zafar, Afia

    2016-02-01

    Early availability of antifungal susceptibilities can ensure timely institution of targeted therapy in candidemia, which can improve patient outcomes. This study prospectively determines the agreement between the results of direct testing of antifungal susceptibilities from blood culture bottles by disk diffusion and Etest and the results of standardized susceptibility testing methods; direct testing would allow susceptibility results to be available 1 to 2 days earlier. A total of 104 blood cultures with different Candida species (28% C. albicans, 27% C. parapsilosis, 26% C. tropicalis, etc.) were evaluated between January 2012 and May 2013 for agreement of fluconazole, voriconazole, and amphotericin B susceptibility results by disk diffusion. Agreement in MICs obtained by Etest was determined for fluconazole (21 isolates), voriconazole (28 isolates), amphotericin (29 isolates), and caspofungin (29 isolates). The kappa scores for categorical agreement were highest for fluconazole by disk diffusion (0.902, standard error [SE] = 0.076) and Etest (1.00, SE = 0.218) and for amphotericin B by disk diffusion (1.00, SE = 0.098). The Pearson correlation (r) of zone diameters was strongest for fluconazole (0.69) and amphotericin (0.70) and moderate for voriconazole (0.60), and the Pearson correlation of MICs was strongest for fluconazole (0.94) and caspofungin (0.88). However, the moderate correlation of amphotericin MICs with zone diameters (-0.42) precludes the use of amphotericin B disk diffusion for susceptibility testing. There were no very major errors; however, there were 1 (1%) major and 5 (4.8%) minor errors with disk diffusion and 4 (13.3%) minor errors with Etest. Thus, antifungal disk diffusion directly from blood culture bottles is a rapid and easy method for fluconazole and voriconazole susceptibility testing for timely tailoring of candidemia therapy. PMID:26607985

  6. Improved Broth Microdilution Method for Antimicrobial Susceptibility Testing of Francisella Noatunensis Orientalis.

    PubMed

    Soto, Esteban; Halliday-Simmonds, Iona; Francis, Stewart; Fraites, Trellor; Martínez-López, Beatriz; Wiles, Judy; Hawke, John P; Endris, Richard D

    2016-09-01

    In this project we optimized a minimal inhibitory concentration testing protocol for Francisella noatunensis orientalis. Thirty-three F. noatunensis orientalis isolates recovered from different fish species and locations were tested, and Escherichia coli ATCC 25922 was used as a quality control reference strain. A modified cation-adjusted Mueller Hinton broth supplemented with 2% IsoVitalex and 0.1% glucose (MMH) was tested at a pH of 6.4 ± 0.1, 7.1 ± 0.1, and 7.3 ± 0.1. Growth curves generated for F. noatunensis orientalis indicated that MMH at a pH of 6.4 ± 0.1 provided optimal growth. There were no significant differences in the growth curves obtained from isolates recovered from different fish species or from fresh or marine water. The pH of 6.4 ± 0.1 in the MMH media interfered with the inhibitory properties of the potentiated sulfonamides (ormetoprim-sulfadimethoxine and trimethoprim-sulfamethoxazole) when using the E. coli ATCC reference strain. Minimal inhibitory concentrations of eight antimicrobials (gentamicin, enrofloxacin, ampicillin, oxytetracycline, erythromycin, florfenicol, flumequine, and oxolinic acid) were similar for all F. noatunensis orientalis isolates. The in vitro susceptibility data provided here can provide a baseline for monitoring the development of antimicrobial resistance among F. noatunensis orientalis isolates, as well as provide valuable data in the development of potential therapeutics. Received October 27, 2015; accepted April 13, 2016. PMID:27484609

  7. Archival-grade optical disc design and international standards

    NASA Astrophysics Data System (ADS)

    Fujii, Toru; Kojyo, Shinichi; Endo, Akihisa; Kodaira, Takuo; Mori, Fumi; Shimizu, Atsuo

    2015-09-01

    Optical discs currently on the market exhibit large variations in life span among discs, making them unsuitable for certain business applications. To assess and potentially mitigate this problem, we performed accelerated degradation testing under standard ISO conditions, determined the probable disc failure mechanisms, and identified the essential criteria necessary for a stable disc composition. With these criteria as necessary conditions, we analyzed the physical and chemical changes that occur in the disc components, on the basis of which we determined technological measures to reduce these degradation processes. By applying these measures to disc fabrication, we were able to develop highly stable optical discs.

  8. Susceptibility to Welding Cracking, Welding Sensitivity, Susceptibility to Welding Seam Cracking, and Test Methods for These Failures

    NASA Technical Reports Server (NTRS)

    Zeyen, K. L.

    1949-01-01

    In the years after the First World war a very rapid development, which even today has by no means come to an end, took place in the field of welding; here also, as in most technical innovations, failures made their appearance. Thereupon comprehensive Investigations were undertaken by steel manufacturers and consumers with cooperation of the authorities. b most cases the reasons for failure could be determined and eliminated and further occurrences avoided by the use of new test methods.

  9. Bile Culture and Susceptibility Testing of Malignant Biliary Obstruction via PTBD

    SciTech Connect

    Yu Haipeng; Guo Zhi Xing Wenge; Guo Xiuying; Liu Fang; Li Baoguo

    2012-10-15

    Purpose: To assess the information obtained by bile culture and susceptibility testing for malignant biliary obstruction by a retrospective one-center study. Methods: A total of 694 patients with malignant biliary obstruction received percutaneous transhepatic biliary drainage during the period July 2003 to September 2010, and subsequently, bile specimens were collected during the procedure. Among the 694 patients, 485 were men and 209 were women, ranging in age from 38 to 78 years (mean age 62 years). Results: A total of 42.9% patients had a positive bile culture (298 of 694). Further, 57 species of microorganisms and 342 strains were identified; gram-positive bacteria accounted for 50.9% (174 of 342) and gram-negative bacteria accounted for 41.5% (142 of 342) of these strains. No anaerobes were obtained by culture during this study. The most common microorganisms were Enterococcus faecalis (41 of 342, 11.9%), Escherichia coli (34 of 342, 9.9%), Klebsiella pneumoniae (28 of 342, 8.2%), Staphylococcus epidermidis (19 of 342, 5.5%), Enterococcus (18 of 342, 5.3%), and Enterobacter cloacae (16 of 342, 4.7%). The percentage of {beta}-lactamase-producing gram-positive bacteria was 27.6% (48 of 174), and the percentage of gram-negative bacteria was 19.7% (28 of 142). The percentage of enzyme-producing Escherichia coli was 61.7% (21 of 34). Conclusion: The bile cultures in malignant biliary obstruction are different from those in the Tokyo Guidelines and other benign biliary obstruction researches, which indicates that a different antibacterial therapy should be applied. Thus, knowledge of the antimicrobial susceptibility data could aid in the better use of antibiotics for the empirical therapy of biliary infection combined with malignant biliary obstruction.

  10. Redundant disc

    NASA Technical Reports Server (NTRS)

    Barack, W. N.; Domas, P. A.; Beekman, S. W. (Inventor)

    1978-01-01

    A rotatable disc is described that consists of parallel plates tightly joined together for rotation about a hub. Each plate is provided with several angularly projecting spaced lands. The lands of each plate are interposed in alternating relationship between the lands of the next adjacent plate. In this manner, circumferential displacement of adjacent sectors in any one plate is prevented in the event that a crack develops. Each plate is redundantly sized so that, in event of structural failure of one plate, the remaining plates support a proportionate share of the load of the failed plate. The plates are prevented from separating laterally through the inclusion of generally radially extending splines which are inserted to interlock cooperating, circumferentially adjacent lands.

  11. Multicenter Evaluation of Fully Automated BACTEC Mycobacteria Growth Indicator Tube 960 System for Susceptibility Testing of Mycobacterium tuberculosis

    PubMed Central

    Bemer, Pascale; Palicova, Frantiska; Rüsch-Gerdes, Sabine; Drugeon, Henri B.; Pfyffer, Gaby E.

    2002-01-01

    The reliability of the BACTEC Mycobacteria Growth Indicator Tube (MGIT) 960 system for testing of Mycobacterium tuberculosis susceptibility to the three front-line drugs (isoniazid [INH], rifampin [RIF], and ethambutol [EMB]) plus streptomycin (STR) was compared to that of the BACTEC 460 TB system. The proportion method was used to resolve discrepant results by an independent arbiter. One hundred and ten strains were tested with an overall agreement of 93.5%. Discrepant results were obtained for seven strains (6.4%) with INH (resistant by BACTEC MGIT 960; susceptible by BACTEC 460 TB), for one strain (0.9%) with RIF (resistant by BACTEC MGIT 960; susceptible by BACTEC 460 TB), for seven strains (6.4%) with EMB (six resistant by BACTEC MGIT 960 and susceptible by BACTEC 460 TB; one susceptible by BACTEC MGIT 960 and resistant by BACTEC 460 TB), and for 19 strains (17.3%) with STR (resistant by BACTEC MGIT 960 and susceptible by BACTEC 460 TB). After resolution of discrepant results, the sensitivity of the BACTEC MGIT 960 system was 100% for all four drugs and specificity ranged from 89.8% for STR to 100% for RIF. Turnaround times were 4.6 to 11.7 days (median, 6.5 days) for BACTEC MGIT 960 and 4.0 to 10.0 days (median, 7.0 days) for BACTEC 460 TB. These data demonstrate that the fully automated and nonradiometric BACTEC MGIT 960 system is an accurate method for rapid susceptibility testing of M. tuberculosis. PMID:11773109

  12. National Committee for Clinical Laboratory Standards agar dilution susceptibility testing of anaerobic gram-negative bacteria.

    PubMed Central

    Brown, W J

    1988-01-01

    One hundred nine recent clinical isolates of anaerobic gram-negative bacteria were tested in triplicate by the National Committee for Clinical Laboratory Standards agar dilution procedure for their susceptibility to 32 antimicrobial agents. All isolates were inhibited by imipenem, but there were significant numbers of strains resistant to other beta-lactam drugs, and therefore the in vitro response to these antimicrobial agents cannot be predicted. This was particularly true for the bile-resistant or Bacteroides fragilis group. beta-Lactamase production was detected in 82% of the bacteroides with the nitrocefin test. Clavulanic acid combined with amoxicillin and ticarcillin and sulbactam combined with ampicillin resulted in synergistic activity against all beta-lactamase-positive organisms. Ceftizoxime was the most active of the cephalosporins. Two percent of the isolates were resistant to chloramphenicol and metronidazole. Clindamycin resistance was detected in 38% of the B. fragilis group, which is a marked increase from the 4% detected 10 years ago at this institution. PMID:3364956

  13. An in situ antimicrobial susceptibility testing method based on in vivo measurements of chlorophyll α fluorescence.

    PubMed

    Heliopoulos, Nikolaos S; Galeou, Angeliki; Papageorgiou, Sergios K; Favvas, Evangelos P; Katsaros, Fotios K; Stamatakis, Kostas

    2015-05-01

    Up to now antimicrobial susceptibility testing (AST) methods are indirect and generally involve the manual counting of bacterial colonies following the extraction of microorganisms from the surface under study and their inoculation in a separate procedure. In this work, an in situ, direct and instrumental method for the evaluation and assessment of antibacterial properties of materials and surfaces is proposed. Instead of indirectly determining antibacterial activity using the typical gram(-) test organisms with the subsequent manual colony count or inhibition zone measurement, the proposed procedure, employs photosynthetic gram(-) cyanobacteria deposited directly onto the surface under study and assesses cell proliferation and viability by a quick, accurate and reproducible instrumental chlorophyll fluorescence spectrophotometric technique. In contrast with existing methods of determination of antibacterial properties, it produces high resolution and quantitative results and is so versatile that it could be used to evaluate the antibacterial properties of any compound (organic, inorganic, natural or man-made) under any experimental conditions, depending on the targeted application. PMID:25771834

  14. Are E-test and Vitek2 good choices for tigecycline susceptibility testing when comparing broth microdilution for MDR and XDR Acinetobacter baumannii?

    PubMed

    Grandesso, Stefano; Sapino, Barbara; Amici, Gianpaolo; Mazzucato, Sandra; Solinas, Maria; Gion, Massimo

    2014-10-01

    This study reports the results of antimicrobial susceptibility testing of 10 MDR and 74 XDR Acinetobacter bauman- nii clinical isolates from our hospital routine. We used three different methods: two automated systems (Sensititre and VITEK 2) and one standardized manual method (E-test). Since many published papers refer to in vitro tests performed by E-test, the aim of this study was to test if this method is reliable for testing tigecycline. The results obtained show that E-test significantly overestimates the MIC of the broth microdilution (reference test), thus ob- taining a significant number of major errors (resistant instead of sensitive). VITEK 2 also shows the same problem, but it is less critical. We therefore conclude that these methods do not seem to be very reliable in the performance of susceptibility testing of MDR and XDR Acinetobacter baumannii against tigecycline. PMID:25387287

  15. Intervertebral disc disease.

    PubMed

    Simpson, S T

    1992-07-01

    This article describes the functional anatomy of intervertebral discs and their relationship to the vertebrae and spinal cord. The pathologic events and clinical complications of intervertebral disc disease are described. A discussion of proper staging of disc disease and appropriate conservative management of degenerative disc disease is included. PMID:1641922

  16. Patient and provider attitudes toward genomic testing for prostate cancer susceptibility: a mixed method study

    PubMed Central

    2013-01-01

    Background The strong association between family history and prostate cancer (PCa) suggests a significant genetic contribution, yet specific highly penetrant PCa susceptibility genes have not been identified. Certain single-nucleotide-polymorphisms have been found to correlate with PCa risk; however uncertainty remains regarding their clinical utility and how to best incorporate this information into clinical decision-making. Genetic testing is available directly to consumers and both patients and healthcare providers are becoming more aware of this technology. Purchasing online allows patients to bypass their healthcare provider yet patients may have difficulty interpreting test results and providers may be called upon to interpret results. Determining optimal ways to educate both patients and providers, and strategies for appropriately incorporating this information into clinical decision-making are needed. Methods A mixed-method study was conducted in Utah between October 2011 and December 2011. Eleven focus group discussions were held and surveys were administered to 23 first-degree relatives of PCa patients living in Utah and 24 primary-care physicians and urologists practicing in Utah to present specific information about these assessments and determine knowledge and attitudes regarding health implications of using these assessments. Results Data was independently coded by two researchers (relative Kappa = .88; provider Kappa = .77) and analyzed using a grounded theory approach. Results indicated differences in attitudes and behavioral intentions between patient and provider. Despite the test’s limitations relatives indicated interest in genetic testing (52%) while most providers indicated they would not recommend the test for their patients (79%). Relatives expected providers to interpret genetic test results and use results to provide personalized healthcare recommendations while the majority of providers did not think the information would be useful in

  17. Prenatal SNP array testing in 1000 fetuses with ultrasound anomalies: causative, unexpected and susceptibility CNVs.

    PubMed

    Srebniak, Malgorzata I; Diderich, Karin Em; Joosten, Marieke; Govaerts, Lutgarde Cp; Knijnenburg, Jeroen; de Vries, Femke At; Boter, Marjan; Lont, Debora; Knapen, Maarten Fcm; de Wit, Merel C; Go, Attie Tji; Galjaard, Robert-Jan H; Van Opstal, Diane

    2016-05-01

    To evaluate the diagnostic value of single-nucleotide polymorphism (SNP) array testing in 1033 fetuses with ultrasound anomalies we investigated the prevalence and genetic nature of pathogenic findings. We reclassified all pathogenic findings into three categories: causative findings; unexpected diagnoses (UD); and susceptibility loci (SL) for neurodevelopmental disorders. After exclusion of trisomy 13, 18, 21, sex-chromosomal aneuploidy and triploidies, in 76/1033 (7.4%) fetuses a pathogenic chromosome abnormality was detected by genomic SNP array: in 19/1033 cases (1.8%) a microscopically detectable abnormality was found and in 57/1033 (5.5%) fetuses a pathogenic submicroscopic chromosome abnormality was detected. 58% (n=44) of all these pathogenic chromosome abnormalities involved a causative finding, 35% (n=27) a SL for neurodevelopmental disorder, and 6% (n=5) a UD of an early-onset untreatable disease. In 0.3% of parental samples an incidental pathogenic finding was encountered. Our results confirm that a genomic array should be the preferred first-tier technique in fetuses with ultrasound anomalies. All UDs involved early-onset diseases, which is beneficial for the patients to know. It also seems that UDs occur at a comparable frequency among microscopic and submicroscopic pathogenic findings. SL were more often detected than in pregnancies without ultrasound anomalies. PMID:26328504

  18. Effect of buffers on testing of Candida species susceptibility to flucytosine.

    PubMed Central

    MacKerrow, S D; Merry, J M; Hoeprich, P D

    1987-01-01

    Synthetic amino acid medium for fungi (SAAMF) is a totally defined, nutritionally adequate, macromolecule-free culture medium for fungi that is buffered with an organic weak acid-weak base pair: 2-(N-morpholino)-propanesulfonic acid (MOPS) and 2-amino-2-(hydroxymethyl)-1,3-propanediol (Tris). In 1984, it was reported that MOPS-Tris in SAAMF antagonized the activity of flucytosine against Candida albicans (D. L. Calhoun and J. N. Galgiani, Antimicrob. Agents Chemother. 26:364-367, 1984). Accordingly, we evaluated the buffering capacity of seven synthetic organic buffers and monobasic potassium phosphate, both singly and in pairs, over the pH range 7.4 to 6.0. Of these buffers, MOPS, BES [N,N-bis(2-hydroxyethyl)-2-aminomethanesulfonic acid], a BES-MOPS combination, and KH2PO4 provided the best buffering. Growth of C. albicans, in unbuffered SAAMF was equivalent overall to that in SAAMF containing buffers, singly or in pairs. Twelve strains of C. albicans and five strains of Candida lusitaniae were tested for susceptibility to flucytosine in SAAMF, with and without buffers. In the presence of Tris, the geometric mean MICs were 6.5- and 3.6-fold higher, respectively, for C. albicans and C. lusitaniae. We recommend replacing Tris with the nonantagonistic MOPS. PMID:3294891

  19. A Low Cost/Low Power Open Source Sensor System for Automated Tuberculosis Drug Susceptibility Testing

    PubMed Central

    Kim, Kyukwang; Kim, Hyeong Keun; Lim, Hwijoon; Myung, Hyun

    2016-01-01

    In this research an open source, low power sensor node was developed to check the growth of mycobacteria in a culture bottle with a nitrate reductase assay method for a drug susceptibility test. The sensor system reports the temperature and color sensor output frequency change of the culture bottle when the device is triggered. After the culture process is finished, a nitrite ion detecting solution based on a commercial nitrite ion detection kit is injected into the culture bottle by a syringe pump to check bacterial growth by the formation of a pigment by the reaction between the solution and the color sensor. Sensor status and NRA results are broadcasted via a Bluetooth low energy beacon. An Android application was developed to collect the broadcasted data, classify the status of cultured samples from multiple devices, and visualize the data for the end users, circumventing the need to examine each culture bottle manually during a long culture period. The authors expect that usage of the developed sensor will decrease the cost and required labor for handling large amounts of patient samples in local health centers in developing countries. All 3D-printerable hardware parts, a circuit diagram, and software are available online. PMID:27338406

  20. A Low Cost/Low Power Open Source Sensor System for Automated Tuberculosis Drug Susceptibility Testing.

    PubMed

    Kim, Kyukwang; Kim, Hyeong Keun; Lim, Hwijoon; Myung, Hyun

    2016-01-01

    In this research an open source, low power sensor node was developed to check the growth of mycobacteria in a culture bottle with a nitrate reductase assay method for a drug susceptibility test. The sensor system reports the temperature and color sensor output frequency change of the culture bottle when the device is triggered. After the culture process is finished, a nitrite ion detecting solution based on a commercial nitrite ion detection kit is injected into the culture bottle by a syringe pump to check bacterial growth by the formation of a pigment by the reaction between the solution and the color sensor. Sensor status and NRA results are broadcasted via a Bluetooth low energy beacon. An Android application was developed to collect the broadcasted data, classify the status of cultured samples from multiple devices, and visualize the data for the end users, circumventing the need to examine each culture bottle manually during a long culture period. The authors expect that usage of the developed sensor will decrease the cost and required labor for handling large amounts of patient samples in local health centers in developing countries. All 3D-printerable hardware parts, a circuit diagram, and software are available online. PMID:27338406

  1. CLSI broth microdilution method for testing susceptibility of Malassezia pachydermatis to thiabendazole

    PubMed Central

    Nascente, Patrícia da Silva; Meinerz, Ana Raquel Mano; de Faria, Renata Osório; Schuch, Luiz Filipe Damé; Meireles, Mário Carlos Araújo; de Mello, João Roberto Braga

    2009-01-01

    Thiabendazole, classified as antiparasitic and also used as an antifungal drug, can be found as otological solution indicated for treatment of parasitic and fungal external otitis in small animals. Malassezia pachydermatis is a yeast recognized as a normal inhabitant on the skin and mucous membranes of dogs and cats. However, it is considered an opportunistic agent that causes external otitis and dermatitis in these animals. The aim of this study was to evaluate the in vitro effect of thiabendazole against 51 isolates of M. pachydermatis using the CLSI Broth Microdilution method that has been adapted for this yeast species (NCCLS, 2002). Based on this test, the Minimum Inhibitory Concentrations (MIC) of thiabendazol was calculated. Subsequently, the susceptibility of each isolate against this antifungal was determined. It was observed that the MIC of thiabendazole against M. pachydermatis ranged from 0.03 to > 4 µg/mL. A total of 13.7% of the isolates were found to be resistant, 47.1% were intermediate and 39.2% were sensitive to the drug. The rate of resistance of the yeasts against thiabendazole was similar to the results previously obtained with other antifungals, while the adapted broth microdilution technique used in this study proved to be efficient. PMID:24031347

  2. Evaluation of Vitek2 and BD Phoenix in antimicrobial susceptibility testing of Acinetobacter baumannii and Pseudomonas aeruginosa.

    PubMed

    Jekarl, Dong Wook; Han, Sang Bong; Kim, Yoon Joo; Shin, Sang Hyun; Park, Kang Gyun; Park, Jung Jun; Han, Kyungja; Park, Yeon-Joon

    2010-08-01

    The accuracy of antimicrobial susceptibility testing of Vitek2 and BD Phoenix against Acinetobacter baumannii and Pseudomonas aeruginosa was evaluated. Both systems showed overall categoric agreement of < or =90% for cefepime and ceftazidime against A. baumannii and imipenem and cefepime (and ceftazidime with Vitek2) against P. aeruginosa because of high minor error rates. PMID:20638609

  3. Broken discs: warp propagation in accretion discs

    NASA Astrophysics Data System (ADS)

    Nixon, Christopher J.; King, Andrew R.

    2012-04-01

    We simulate the viscous evolution of an accretion disc around a spinning black hole. In general, any such disc is misaligned, and warped by the Lense-Thirring effect. Unlike previous studies, we use effective viscosities constrained to be consistent with the internal fluid dynamics of the disc. We find that non-linear fluid effects, which reduce the effective viscosities in warped regions, can promote breaking of the disc into two distinct planes. This occurs when the Shakura & Sunyaev dimensionless viscosity parameter α is ≲0.3 and the initial angle of misalignment between the disc and hole is ≳45°. The break can be a long-lived feature, propagating outwards in the disc on the usual alignment time-scale, after which the disc is fully co-aligned or counter-aligned with the hole. Such a break in the disc may be significant in systems where we know the inclination of the outer accretion disc to the line of sight, such as some X-ray binaries: the inner disc, and so any jets, may be noticeably misaligned with respect to the orbital plane.

  4. Disc-planet interactions in subkeplerian discs

    NASA Astrophysics Data System (ADS)

    Paardekooper, S.-J.

    2009-11-01

    Context: One class of protoplanetary disc models, the X-wind model, predicts strongly subkeplerian orbital gas velocities, a configuration that can be sustained by magnetic tension. Aims: We investigate disc-planet interactions in these subkeplerian discs, focusing on orbital migration for low-mass planets and gap formation for high-mass planets. Methods: We use linear calculations and nonlinear hydrodynamical simulations to measure the torque and look at gap formation. In both cases, the subkeplerian nature of the disc is treated as a fixed external constraint. Results: We show that, depending on the degree to which the disc is subkeplerian, the torque on low-mass planets varies between the usual type I torque and the one-sided outer Lindblad torque, which is also negative but an order of magnitude stronger. In strongly subkeplerian discs, corotation effects can be ignored, making migration fast and inward. Gap formation near the planet's orbit is more difficult in such discs, since there are no resonances close to the planet accommodating angular momentum transport. The location of the gap is shifted inwards with respect to the planet, leaving the planet on the outside of a surface density depression. Conclusions: Depending on the degree to which a protoplanetary disc is subkeplerian, disc-planet interactions can be very different from the usual Keplerian picture, making these discs in general more hazardous for young planets.

  5. Accuracy of automated and manual systems for susceptibility testing of Pseudomonas aeruginosa to piperacillin and piperacillin-tazobactam.

    PubMed

    Gagliotti, Carlo; Sarti, Mario; Sabia, Carla; Gargiulo, Raffaele; Rossolini, Gian Maria; Carillo, Carmelina; Cassani, Carla; Cipolloni, Antonio Paolo; Pedna, Federica; Rossi, Maria Rita; Incerti, Silvia Storchi; Testa, Giovanna; Venturelli, Claudia; Moro, Maria Luisa

    2011-01-01

    This study aimed to evaluate the accuracy of routine systems (Vitek2 cards AST-N022 and AST-N026; Kirby Bauer; Etest) for susceptibility testing of Pseudomonas aeruginosa to piperacillin and piperacillin-tazobactam. Vitek2 (card AST-N022) showed the worst performance; the other three methods (Vitek2 card AST-N026, Kirby-Bauer and E-test) performed comparably but never fulfilled the minimal standard proposed by FDA. PMID:21344153

  6. [Recommendations from MENSURA for selection of antimicrobial agents for susceptibility testing and criteria for the interpretation of antibiograms].

    PubMed

    2000-03-01

    This document includes the recommendations from the Spanish antibiogram committee (The MENSURA group, Mesa Española de Normalización de la Sensibilidad y Resistencia a los Antimicrobianos, under the auspices of the Sociedad Española de Quimioterapia and Sociedad Española de Enfermedades Infecciosas y Microbiología Clínica) for the selection of antimicrobials for susceptibility testing. Separate tables for each group of organism with proposed susceptibility and resistance breakpoints are updated and comparatively presented with those of other groups, such us NCCLS, CA-SFM and BSAC. The susceptibility breakpoint tends to identify the fully susceptible population, which probably lacks any specific resistance mechanism. The analysis of MIC distributions for different homogeneous populations (same species) is used to define breakpoints for susceptibility. The resistance breakpoint is based on pharmacological and clinical data obtained when the corresponding antibiotic is administered with a conventional schedule. The primary objective of the Spanish MENSURA group is to contribute to the international consensus on the establishment of breakpoints. PMID:10856132

  7. Carbapenem Resistance in Klebsiella pneumoniae Not Detected by Automated Susceptibility Testing

    PubMed Central

    Kalsi, Rajinder K.; Williams, Portia P.; Carey, Roberta B.; Stocker, Sheila; Lonsway, David; Rasheed, J. Kamile; Biddle, James W.; McGowan, John E.; Hanna, Bruce

    2006-01-01

    Detecting β-lactamase–mediated carbapenem resistance among Klebsiella pneumoniae isolates and other Enterobacteriaceae is an emerging problem. In this study, 15 blaKPC-positive Klebsiella pneumoniae that showed discrepant results for imipenem and meropenem from 4 New York City hospitals were characterized by isoelectric focusing; broth microdilution (BMD); disk diffusion (DD); and MicroScan, Phoenix, Sensititre, VITEK, and VITEK 2 automated systems. All 15 isolates were either intermediate or resistant to imipenem and meropenem by BMD; 1 was susceptible to imipenem by DD. MicroScan and Phoenix reported 1 (6.7%) and 2 (13.3%) isolates, respectively, as imipenem susceptible. VITEK and VITEK 2 reported 10 (67%) and 5 (33%) isolates, respectively, as imipenem susceptible. By Sensititre, 13 (87%) isolates were susceptible to imipenem, and 12 (80%) were susceptible to meropenem. The VITEK 2 Advanced Expert System changed 2 imipenem MIC results from >16 μg/mL to <2 μg/mL but kept the interpretation as resistant. The recognition of carbapenem-resistant K. pneumoniae continues to challenge automated susceptibility systems. PMID:16965699

  8. Antimicrobial potentials of Catharanthus roseus by disc diffusion assay.

    PubMed

    Bakht, Jehan; Syed, Fatema; Shafi, Mohammad

    2015-05-01

    The present research work investigates the in vitro antimicrobial activity of different solvent extracted samples from the aerial parts (stem, leaf, fruit and flower) of C. roseus against different microbial species using disc diffusion assay at two different concentrations of 1 and 2 mg disc-1. Hexane extracted samples inhibited the growth of all tested microbial strains except S. typhi. Similarly, ethyl acetate extracted samples was effective to control the activity of all the tested microbial strains. E. coli and S. typhi showed resistance to chloroform extracted samples and the remaining eight microbial strains were susceptible to the same extract. Butanol extracted samples did not inhibit the growth of K. pneumonia and S. typhi at low concentration, however, at higher concentration the same extract reduced the growth of different microbes. Methanol extracted samples effectively controlled the growth of all tested microbes at both concentrations except for S. typhi. Water extracted samples did not inhibit the growth at low concentration except E. coli, K. pneumonia and S. aureus and were ineffective against P. aeroginosa at both concentration. C. albicans, showed resistance against chloroform and water extracted samples at low concentration and susceptible to other solvent extracted samples at both concentration. All fractions were effective against plant pathogens i.e. E. carotovora and A. tumefaciens. PMID:26004715

  9. Antimicrobial susceptibility testing for Helicobacter pylori isolates from Brazilian children and adolescents: Comparing agar dilution, E-test, and disk diffusion

    PubMed Central

    Ogata, Silvio Kazuo; Gales, Ana Cristina; Kawakami, Elisabete

    2014-01-01

    Antimicrobial susceptibility testing for Helicobacter pylori is increasingly important due to resistance to the most used antimicrobials agents. Only agar dilution method is approved by CLSI, but it is difficult to perform routinely. We evaluated the reliability of E-test and disk diffusion comparing to agar dilution method on Helicobacter pylori antimicrobial susceptibility testing. Susceptibility testing was performed for amoxicillin, clarithromycin, furazolidone, metronidazole and tetracycline using E-test, disk-diffusion and agar dilution method in 77 consecutive Helicobacter pylori strains from dyspeptic children and adolescents. Resistance rates were: amoxicillin - 10.4%, 9% and 68.8%; clarithromycin - 19.5%, 20.8%, 36.3%; metronidazole - 40.2%33.7%, 38.9%, respectively by agar dilution, E-test and disk diffusion method. Furazolidone and tetracycline showed no resistance rates. Metronidazole presented strong correlation to E-test (r = 0.7992, p < 0.0001) and disk diffusion method (r=-0.6962, p < 0.0001). Clarithromycin presented moderate correlation to E-test (r = 0.6369, p < 0.0001) and disk diffusion method (r=−0.5656, p < 0.0001). Amoxicillin presented weak correlation to E-test (r = 0.3565, p = 0.0015) and disk diffusion (r=−0.3565, p = 0.0015). Tetracycline presented weak correlation with E-test (r = 0.2346, p = 0.04) and furazolidone to disk diffusion (r=−0.0288, p = 0.8038). E-test presented better agreement with gold standard. It is an easy and reliable method for Helicobacter pylori susceptibility testing. Disk diffusion method presented high disagreement and high rates of major errors. PMID:25763052

  10. Photographic and luminometric detection of luciferase reporter phages for drug susceptibility testing of clinical Mycobacterium tuberculosis isolates.

    PubMed

    Hazbón, Manzour Hernando; Guarín, Nora; Ferro, Beatriz Eugenia; Rodríguez, Ana Lucía; Labrada, Luz Angela; Tovar, Rafael; Riska, Paul F; Jacobs, William R

    2003-10-01

    Luciferase reporter phages (LRPs) have proven to be efficient tools for drug susceptibility testing of Mycobacterium tuberculosis. Luminometric detection of LRP activity offers higher sensitivity and quantitative results, while a Polaroid film detection method offers a "low-tech" inexpensive alternative that is called the Bronx box. In this work we evaluated, improved, and compared the performance of the luminometer and the Bronx box formats for drug susceptibility testing with LRPs by using 51 clinical isolates of M. tuberculosis, with the agar proportion method (PM) serving as reference. The sensitivity in detecting resistance to isoniazid and rifampin, antibiotics that define multidrug resistance (MDR), was 100% for both methods. The turnaround time for results was reduced from 3 weeks for PM to 54 or 94 h for luminometry or the Bronx box, respectively. These results support the utility of LRPs as a screening test for the surveillance of MDR tuberculosis. PMID:14532245