Visual Detection of rpoB Mutations in Rifampin-Resistant Mycobacterium tuberculosis Strains by Use of an Asymmetrically Split Peroxidase DNAzyme
Visual Detection of rpoB Mutations in Rifampin-Resistant Mycobacterium tuberculosis Strains by Use of an Asymmetrically Split Peroxidase DNAzyme
复制标题
使用不对称裂解过氧化物酶脱氧核糖核酸酶目视检测利福平耐药结核分枝杆菌菌株中的 rpoB 突变
DOI：
10.1128/jcm.01292-12
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发表时间：
2012-11-01
影响因子：
9.4
通讯作者：
Zhang, Xiao-Lian
中科院分区：
文献类型：
--
作者：
Deng, Minggang;Feng, Shuo;Zhang, Xiao-Lian
摘要耐多药结核分枝杆菌对异烟肼和利福平这两种一线抗结核药物耐药，导致结核病复发。M.结核病生长缓慢，传统的检测方法检测其耐药性需要时间，不能满足临床需要。DNA探针的使用使得检测利福平耐药性成为可能。我们建立了一种检测耐利福平分枝杆菌耐药突变的不对称分裂组装DNA过氧化物酶法。结核病rpoB基因迅速和明显。同时设计了一种新的策略，以消除目标DNA复杂的二级结构对探针检测的不利影响，提高探针的检测效率。该检测体系由5组检测组成，覆盖rpoB基因的利福平耐药决定区，检测40种突变，包括最常见的密码子531和526处突变。每一组检测或单个突变等位基因检测可以区分相应的突变DNA序列和野生型DNA序列。
ABSTRACT Multidrug-resistant Mycobacterium tuberculosis is resistant to two first-line antituberculosis drugs, isoniazid and rifampin, resulting in the relapse of tuberculosis. M. tuberculosis grows very slowly, and thus traditional examination methods take time to test its drug resistance and cannot meet clinical needs. The use of a DNA probe makes it possible to test rifampin resistance. We developed an asymmetrical split-assembly DNA peroxidase assay to detect drug-resistant mutation of rifampin-resistant M. tuberculosis in the rpoB gene rapidly and visibly. A new strategy was also designed to eliminate the adverse effects caused by the complicated secondary structure of the target DNA and to improve the efficiency of the probes. This detection system consists of five group detections, covers rifampin-resistant determination region of the rpoB gene, and tests 40 kinds of mutations, including the most common mutations at codons 531 and 526. Every group detection or individual mutant allele detection can distinguish corresponding mutant DNA sequences from the wild-type DNA sequences.