High resolution melting curve assay for rapid detection of drug-resistant Mycobacterium tuberculosis

被引：18
|
作者：
Nagai, Yuhki [1 ]
Iwade, Yoshito [1 ]
Hayakawa, Eri [2 ]
Nakano, Manabu [2 ]
Sakai, Takashi [2 ]
Mitarai, Satoshi [3 ]
Katayama, Masahiko [1 ]
Nosaka, Tetsuya [4 ]
Yamaguchi, Tetsuo [1 ]
机构：
[1] Mie Prefecture Hlth & Environm Res Inst, Yokaichi, Mie 5121211, Japan
[2] Natl Hosp Org, Mie Chuo Med Ctr, Tsu, Mie 5141101, Japan
[3] Japan AntiTB Assoc, Res Inst TB, Dept Mycobacterium Reference & Res, Kiyose, Tokyo 2048533, Japan
[4] Mie Univ, Grad Sch Med, Dept Microbiol & Mol Genet, Tsu, Mie 5148507, Japan
来源：
JOURNAL OF INFECTION AND CHEMOTHERAPY | 2013年 / 19卷 / 06期
关键词：
Mycobacterium tuberculosis; Drug resistance; High resolution melting (HRM) analysis; REAL-TIME PCR; STRAND CONFORMATION POLYMORPHISM; RPOB MUTATIONS; ETHAMBUTOL RESISTANCE; ISONIAZID RESISTANCE; STREPTOMYCIN RESISTANCE; MOLECULAR ANALYSIS; SEQUENCE-ANALYSIS; EMBB CODON-306; GENE-MUTATIONS;
D O I：
10.1007/s10156-013-0636-3
中图分类号：
R51 [传染病];
学科分类号：
100401 ;
摘要：
We developed and evaluated a high resolution melting (HRM) curve assay by using real-time PCR for the detection of the most frequent mutations of Mycobacterium tuberculosis, which are responsible for the resistance of four anti-TB drugs: rifampicin, isoniazid, ethambutol, and streptomycin. The HRM assay was successfully used for the detection of dominant mutations: A516V, H526A, H526T, S531L, L533P, and A516G/S531L in rpoB; S315T, and S315A in katG; -15C/T, and -8T/C in mab-inhA; M306I in embB; K88Q and K43R in rpsL; and 513A/C in rrs. We were able to discriminate the mutant from the wild type by analyzing the melting-curve shape in 40 clinical M. tuberculosis isolates, and the results of the HRM assay were completely consistent with those of DNA sequencing. This HRM assay is a simple, rapid, and cost-effective method that can be performed in a closed tube. Therefore, our assay is a potentially useful tool for the rapid detection of drug-resistant M. tuberculosis.
引用
收藏
页码：1116 / 1125
页数：10
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