Application of real - time PCR for rapid detection of Rifampicin resistant Mycobacterium tuberculosis

The emergence of drug-resistant strain of M. tuberculosis is one of the most critical issues facing TB researchers and clinicians. rapid diagnosis of drug-resistant tuberculosis is essential for the prompt initiation of effective second-line therapy to improve treatment outcome and limit transmission of this obstinate disease. The aim of this study is to develop a Real-time PCR assay for the detection of mutations in RRDR (rifampcin resistance determinant region) of rpoB which conferring rifampicin resistance in Mycobacterium tuberculosis In this experimental study، the primer and probe set were designed for a RRDR region of rpoB gene using a specialized software. Clinical specimens that had previously been evaluated resistant or sensitive by using convential method، were used for assessing the clinical sensitivity and specificity of the assay. The clinical sensitivity of the assay was determined 100%. The primers and the probes were rpoB specific and no cross-reaction was observed with other microorganisms and human genome bioinformatically. The clinical specificity of developed Real-timePCR assay was examined experimentally using 25 negative samples and determined to be 100%. The developed real-time PCR assay can be used as an appropriate and efficient tool for the rapid detection of rifampicin-resistant Mycobacterium tuberculosis.