Open AccessPerformance of TaqMan array card to detect TB drug resistance on direct specimens
Author(s) -
Sayera Banu,
Suporn Pholwat,
Suporn Foongladda,
Rattapha Chinli,
Duangjai Boonlert,
Sara Sabrina Ferdous,
S. M. Mazidur Rahman,
Ananna Rahman,
Shahriar Ahmed,
Eric R. Houpt
Publication year - 2017
Publication title -
plos one
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.99
H-Index - 332
ISSN - 1932-6203
DOI - 10.1371/journal.pone.0177167
Subject(s) - rpob , mycobacterium tuberculosis , inha , sputum , genotype , taqman , rifampicin , tuberculosis , isoniazid , drug resistance , pyrazinamide , genotyping , biology , sputum culture , virology , darunavir , medicine , microbiology and biotechnology , polymerase chain reaction , genetics , gene , pathology , viral load , human immunodeficiency virus (hiv) , antiretroviral therapy
Culture based phenotypic drug susceptibility testing (DST) for Mycobacterium tuberculosis (TB) is time consuming therefore rapid genotypic methods are increasingly being utilized. We previously developed and evaluated on TB isolates a rapid genotypic TaqMan array card (TAC) that detects mutations in several resistance-associated genes using dozens of primer pairs, probes, and high resolution melt analysis, with >96% accuracy versus Sanger sequencing. In this study we examined the performance of TAC on sputum, comparing results between 71 paired sputum and TB isolates of which 62 were MDR-TB. We also adapted the TAC to include wild-type probes and broadened coverage for rpoB and gyrA mutations. TAC was 89% successful at detecting wild-type or mutations within inhA , katG , rpoB , eis , gyrA , rplC , and pncA on smear positive sputa and 33% successful on smear negative sputa. The overall accuracy of these detections as compared to the TAC results of the paired isolate was 95% ± 7 (average sensitivity 98% ± 3; specificity 92% ± 14). Accuracy of sputum TAC results versus phenotypic DST for isoniazid, rifampin, ofloxacin/moxifloxacin, and pyrazinamide was 85% ± 12. This was similar to that of the isolate TAC results (accuracy 88% ± 13), thus inaccuracies primarily reflected intrinsic genotypic-phenotypic discordance. The TAC is a rapid, modular, comprehensive, and accurate TB DST for the major first and second line TB drugs and could be used for supplemental testing of GeneXpert resistant smear positive sputum.