Development of a single multiplex amplification refractory mutation system PCR for the detection of rifampin-resistant Mycobacterium tuberculosis |
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Authors: | Xiaodan Shi Chen Zhang Ming Shi Mengjie Yang Yi Zhang Ji Wang Hongwei Shen Gang Zhao Xuejun Ma |
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Institution: | 1. Key Laboratory of Medical Virology, Ministry of Health, National Institute for Viral Disease Control and Prevention, Chinese Center for Disease Control and Prevention, Changbai Rd 155, Beijing 102206, China;2. Department of Neurology, Xijing Hospital, The Fourth Military Medical University, Changle-Xi Road 167, Xi''an 710032, China |
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Abstract: | A rapid and simple method for the detection of drug-resistant Mycobacterium tuberculosis is critical for the efficient treatment and control of this pathogen in developing country. Here we developed a single multiplex amplification refractory mutation system (M-ARMS) PCR, in which chimeric-primer and temperature switch PCR (TSP) strategy were included. Using this method, we detected rifampin resistance-associated mutations at codons 511, 516, 526 and 531 in the rifampin resistance-determining region of rpoB gene. The performance of M-ARMS-PCR assay was evaluated with 135 cultured isolates of M. tuberculosis. The sensitivity and specificity were 94.2% and 100%, respectively, compared with direct DNA sequencing, and 86.67% and 89.71%, respectively, compared with culture-based phenotypic drug susceptibility testing. Therefore, this newly-developed M-ARMS-PCR method is useful and efficient with an intended application in provincial Centers for Disease Control and Prevention for rapid detection of rifampin resistance-associated mutations. |
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Keywords: | RRDR rifampicin resistance determining region Mtb Mycobacterium tuberculosis INH isoniazid RIF rifampicin EMB ethambutol SM streptomycin MDR-TB multidrug-resistant tuberculosis DST drug susceptibility testing DHHS Department of Health and Human Services ARMS-PCR amplification refractory mutation system PCR M-ARMS-PCR multiplex amplification refractory mutation system PCR DR drug resistance LJ Lowenstein&ndash Jensen media SDS sodium dodecyl sulfate |
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