Last data update: May 06, 2024. (Total: 46732 publications since 2009)
Records 1-2 (of 2 Records) |
Query Trace: Hooks DP[original query] |
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Evaluation of mycobacterial interspersed repetitive-unit-variable-number tandem-repeat genotyping as performed in laboratories in Canada, France, and the United States
Cowan LS , Hooks DP , Christianson S , Sharma MK , Alexander DC , Guthrie JL , Jamieson FB , Supply P , Allix-Beguec C , Cruz L , Desmond E , Kramer R , Lugo S , Rudrik J . J Clin Microbiol 2012 50 (5) 1830-1 The external quality assessment of 24-locus mycobacterial interspersed repetitive-unit–variable-number tandem-repeat (MIRU-VNTR) genotyping by de Beer et al. reveals issues with its international performance (5). Detailed analysis of the data was confounded by the complexity of the participants. The five genotyping laboratories in Canada and the United States participating in this study use similar typing protocols based on the standardized protocol proposed by Supply et al. (8) and developed in collaboration with each other. Systems for routine handling of samples and data management are well established. Quality control (QC) and assurance measures include routine testing of the Mycobacterium tuberculosis strain H37Rv and repeat testing of 1% of isolates at an external laboratory. The laboratorians conducting the analysis have at a minimum 5 years of experience performing MIRU-VNTR typing. This cohesiveness allows for a more in-depth analysis of the data collected by de Beer et al. | Each laboratory reported 24-locus MIRU-VNTR results for the proficiency testing panel of 30 DNA samples (including 10 pairs of duplicates), and their performance is summarized in Table 1. Reproducibility as calculated at the sample level and disregarding missing results ranged from 93% to 100%, and typeability as calculated by the percentage of loci with a reportable result ranged from 98.9% to 100%. Here we present a detailed description of the 38 observed discrepancies to provide a more complete understanding of the performance of MIRU-VNTR typing in our laboratories. |
Molecular detection of mutations associated with first- and second-line drug resistance compared with conventional drug susceptibility testing of Mycobacterium tuberculosis.
Campbell PJ , Morlock GP , Sikes RD , Dalton TL , Metchock B , Starks AM , Hooks DP , Cowan LS , Plikaytis BB , Posey JE . Antimicrob Agents Chemother 2011 55 (5) 2032-41 The emergence of multi and extensively drug-resistant tuberculosis is a significant impediment to the control of this disease because treatment becomes more complex and costly. Reliable and timely drug susceptibility testing is critical to ensure patients receive effective treatment and become non-infectious. Molecular methods can provide accurate and rapid drug susceptibility results. We used DNA sequencing to detect resistance to the first-line antituberculosis drugs, isoniazid (INH), rifampin (RIF), pyrazinamide (PZA), and ethambutol (EMB), and the second-line drugs, amikacin (AMK), capreomycin (CAP), kanamycin (KAN), ciprofloxacin, (CIP) and ofloxacin (OFX). Nine loci were sequenced: rpoB for resistance to RIF, katG and inhA (INH), pncA (PZA), embB (EMB), gyrA (CIP and OFX), rrs, eis, and tlyA (KAN, AMK, and CAP). A total of 314 clinical M. tuberculosis complex isolates, representing a variety of antibiotic resistance patterns, genotypes and geographical origins were analyzed. The molecular data were compared to the phenotypic data and the accuracy values were calculated. Sensitivity and specificity values (as percentages) for the first-line drug loci were rpoB (97.1, 93.6), katG (85.4, 100), inhA (16.5, 100), katG and inhA together (90.6, 100) pncA (84.6, 85.8), and embB (78.6, 93.1). The values for the second-line drugs were also calculated. The size and scope of this study, in numbers of loci and isolates examined, and the phenotypic diversity of those isolates, support the use of DNA sequencing to detect drug resistance in the M. tuberculosis complex. Further, the results can be used to design diagnostic tests utilizing other mutation detection technologies. |
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