Triplex real-time PCR melting curve analysis for detecting Mycobacterium tuberculosis mutations associated with resistance to second-line drugs in a single reaction.

ABSTRACT:

Objectives

Resistance to anti-tuberculosis (TB) drugs has been a great challenge for global TB control. Limited tools have been developed to detect resistance to second-line drugs that are associated with extensively drug-resistant (XDR) TB. In this study we aimed to develop a simple and widely applicable assay for detecting mutations associated with second-line drug resistance in Mycobacterium tuberculosis.

Methods

Three dually labelled probes targeting gyrA, rrs and the promoter of eis were designed to detect resistance to fluoroquinolones and second-line injectable agents (capreomycin, amikacin and kanamycin). A triplex reaction with all three probes and corresponding primers was first tested against 13 isolates with different mutations in the targeted regions. Then, the

SUBMITTER: Liu Q 

PROVIDER: S-EPMC3625432 | biostudies-literature | 2013 May

REPOSITORIES: biostudies-literature