Publication: Use of mycobacteriophage quantitative PCR on MGIT broths for a rapid tuberculosis antibiogram
Issued Date
2014-01-01
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ISSN
1098660X
00951137
00951137
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2-s2.0-84899529888
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Mahidol University
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SCOPUS
Bibliographic Citation
Journal of Clinical Microbiology. Vol.52, No.5 (2014), 1523-1528
Suggested Citation
Suporn Foongladda, Wiphat Klayut, Rattapha Chinli, Suporn Pholwat, Eric R. Houpt Use of mycobacteriophage quantitative PCR on MGIT broths for a rapid tuberculosis antibiogram. Journal of Clinical Microbiology. Vol.52, No.5 (2014), 1523-1528. doi:10.1128/JCM.03637-13 Retrieved from: https://repository.li.mahidol.ac.th/handle/20.500.14594/34614
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Title
Use of mycobacteriophage quantitative PCR on MGIT broths for a rapid tuberculosis antibiogram
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Abstract
Phenotypic culture-based drug susceptibility testing (DST) for Mycobacterium tuberculosis is a valuable tool to identify four to six active drugs for individualized multidrug-resistant (MDR) tuberculosis (TB) regimens. Current culture-based methods are slow, however; therefore, we evaluated a rapid mycobacteriophage-based quantitative PCR (qPCR) assay for use directly on M. tuberculosis-positive MGIT broths. We compared phage qPCRs, using a simple cutoff of 3 for the ΔCq value (where Cq is quantification cycle, and ΔCq is calculated as the Cq of starting phage minus the Cq of TB isolates in drug-containing medium), on 325 clinical M. tuberculosis MGIT broth cultures versus the respective subcultured isolates tested by agar proportion. The median accuracy for the 13 drugs/concentrations tested was 98%, with most discrepancies being false-resistant results. Evaluation of phage qPCR on greater numbers of resistant strains of 393 isolates grown on Löwenstein-Jensen medium showed similar findings, with a median accuracy, sensitivity, and specificity of 97%, 90%, and 99%, respectively. This rapid culture-based DST methodology can be performed for any drug on TB-positive MGIT broths, with a specimen-to-antibiogram turnaround time of approximately 23.9 days, compared with waiting 58.6 days for isolate growth on solid medium followed by agar proportion DST. © 2014, American Society for Microbiology. All Rights Reserved.