Abstract
The emergence of multidrug-resistant tuberculosis (MDR-TB) and extensively drug-resistant tuberculosis (XDR-TB) strains is a major health problem for high Tuberculosis (TB) incidence countries. Therefore, it is of interest to identify antibiotic resistant bacteria by mismatch detection using DNA hybridization. We generated PCR products for five genes (rpoB, inhA, katG, gyrA and rrs) associated with drug resistance TB from MDR and XDR Mycobacterium tuberculosis (MTB) DNA samples. These were hybridized to PCR products from MTB H37Rv (pansusceptible laboratory strain) to generate DNA hetero-duplex products, which was digested by Detection Enzyme (GeneArt Genomic Cleavage Detection Kit) and visualized by agarose gel electrophoresis. Results show different bands with sizes of 400 bp and 288 bp (rpoB), 280 bp (inhA), 310 bp (katG), 461 bp (gyrA) and 427 bp (rrs) suggesting mutations in DNA heteroduplex for each gene. Detection Enzyme specifically cleaves DNA hetero-duplex with mismatch. The technique helps in the improved detection of MDR (mutations in rpoB, inhA and katG) and XDR (mutations in rpoB, inhA katG, gyrA and rrs) MTB strains. Moreover, the technique is customized without expensive specialized equipment to detect mutations. It is also fast, efficient and easy to implement in standard molecular biology laboratories.
Highlights
In 2016, there were an estimated 10.4 million incident cases of TB [1] .The estimated global burden of multidrug-resistant tuberculosis (MDR-TB) is 500 000 patients and approximately half of MDR-TB new cases are found in China, India and Russia [2]
It is of interest to identify antibiotic resistant bacteria by mismatch mutation detections in gene fragments associated to drug resistance in Mycobacterium tuberculosis (MTB) samples
We hybridized five DNA fragments from M. tuberculosis samples to detect mutations in genes associated with drug resistance
Summary
In 2016, there were an estimated 10.4 million incident cases of TB (range, 8.8 million to 12.2 million) [1] .The estimated global burden of MDR-TB is 500 000 patients and approximately half of MDR-TB new cases are found in China, India and Russia [2]. Different studies indicate that mutations in rpoB, inhA, katG, gyrA and rrs MTB genes confer drug resistance to rifampicin (RIF), isoniazid (INH), fluoroquinolone (FLQ) and aminoglycosides (AMG), respectively [4]. GyrA mutations account for more than 90% of drug resistance to FLQs. Mutations in rrs gene correlate phenotypically with high levels of resistance to KAN, AMK and CAP [6] in specific regions.
Sign-in/Register to view highlights & summary Sign-in/Register to access full text options 
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a callDisclaimer: All third-party content on this website/platform is and will remain the property of their respective owners and is provided on "as is" basis without any warranties, express or implied. Use of third-party content does not indicate any affiliation, sponsorship with or endorsement by them. Any references to third-party content is to identify the corresponding services and shall be considered fair use under The CopyrightLaw.
