Allele Specific-PCR method for rapid detection of gyrA gene mutaions in clinical isolates of Mycobacterium tuberculosis

• Main Authors: Vahideh Vahidi, Mohammad Reza Zolfaghari, Azam Ahmadi, Mana Shojapour, Seyed Reza Moadab, Mohammad Arjomandzadegan
• Format: Article
• Language: English
• Published: University of Isfahan 2014-04-01
• Series: Biological Journal of Microorganism
• Subjects: Mycobacterium tuberculosis; Allele Specific-PCR; Ofloxacin
• Online Access: http://uijs.ui.ac.ir/bjm/browse.php?a_id=59&slc_lang=en&sid=1&ftxt=1

Introduction: Resistance to Fluroquinolones drugs are increasingly expanded. A molecular method was designed and compared for rapid detection of resistance to ofloxacin in clinical isolates of Mycobacterium tuberculosis. Materials and methods: From 136 M. tuberculosis clinical isolates, 41 strains were used for comparison of detection of mutations associated with resistance in gyrA gene by Allele Specific-PCR (AS PCR). Specific internal primers were designed for detecting any changes in 90, 91 and 94 codons. Sequencing method was accomplished for evaluation of the results as gold standard. Results: AS-PCR method could detect mutations by formation or not formation of internal bounds and had good performance. Totally, from 37 strains phenotypically resistant to ofloxacine 32 strains were mutant and 5 strains were non mutant that have Sensitivity and specificity, 86/11% and 100%, respectively. Sequence results were concordant by results molecular methods. Discussion and conclusion: Results of the study showed that AS-PCR method could be used as a routine test for fast detection of resistance to fluroquinolones in M. tuberculosis.