Detection of Mycobacterium tuberculosis resistance mutations to rifampin and isoniazid by real-time PCR

Indian Journal of Medical Microbiology
A HristeaA Streinu-Cercel

Abstract

The objective of our study was to evaluate the use of a real-time polymerase chain reaction (PCR)-based technique for the prediction of phenotypic resistance of Mycobacterium tuberculosis. We tested 67 M tuberculosis strains (26 drug resistant and 41 drug susceptible) using a method recommended for the LightCycler platform. The susceptibility testing was performed by the absolute concentration method. For rifampin resistance, two regions of the rpoB gene were targeted, while for identification of isoniazid resistance, we searched for mutations in katG and inhA genes. The sensitivity and specificity of this method for rapid detection of mutations for isoniazid resistance were 96% (95% CI: 88% to 100%) and 95% (95% CI: 89% to 100%), respectively. For detection of rifampin resistance, the sensitivity and specificity were 92% (95% CI: 81% to 100%) and 74% (95% CI: 61% to 87%), respectively. The main isoniazid resistance mechanism identified in our isolates is related to changes in the katG gene that encodes catalase. We found that for rifampin resistance the concordance between the predicted and observed phenotype was less than satisfactory. Using this method, the best accuracy for genotyping compared with phenotypic resistance tes...Continue Reading

References

Jul 1, 1997·Antimicrobial Agents and Chemotherapy·W H HaasH J Bremer
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Citations

May 17, 2011·FEMS Immunology and Medical Microbiology·Laila NimriRaymond Batchoun
Jan 14, 2011·The New England Journal of Medicine·Norbert HeinrichMichael Hoelscher
Apr 12, 2021·Methods : a Companion to Methods in Enzymology·Anastasiya KostyushevaVladimir Chulanov

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