Zhang S-L, Shen J-G, Xu P-H, Li D-X, Sun Z-Q, Li L, Yang Z-R, Sun Q
College of Life Sciences, Key Laboratory of Bio-resource and Bio-control, Sichuan University, Chengdu, Sichuan, and Research Center for Tuberculosis, Henan Chest Hospital, Zhengzhou, Henan, China.
J Appl Microbiol. 2007 Oct;103(4):1262-71. doi: 10.1111/j.1365-2672.2007.03350.x.
To develop and evaluate a novel genotypic test for rapid detection of rifampicin and isoniazid resistance of multidrug-resistant (MDR) Mycobacterium tuberculosis isolates by a multiplex probe array.
A multiplex probe array was designed for genotypic test to simultaneously screen the mutations of rpoB, katG, inhA and ahpC genes, associated with rifampin and isoniazid resistance in M. tuberculosis, with a probe detecting one of the recently confirmed genetic markers of isoniazid resistance ahpC-6 and -9 locus added. By using the genotypic test developed, 52 MDR isolates were identified, among which 46 isolates had mutations in rpoB (88.5%) and 45 at codon 315 of katG, regulatory region of inhA and oxyR-ahpC intergenic region (86.5%), whereas all 35 susceptible isolates identified showed a wild-type hybridization pattern. The sensitivity and specificity were 88.5% and 100% for rifampicin resistance, and 86.5% and 100% for isoniazid resistance, respectively.
A rapid and simultaneous detection of rifampicin and isoniazid resistance caused by the mutations of rpoB, katG, inhA and ahpC genes in M. tuberculosis isolates could be achieved by a multiplex probe array developed.
This genotypic test protocol has the potential to be developed on clinical application for the rapid detection of drug resistant M. tuberculosis isolates before an efficient chemotherapy is initiated.
开发并评估一种新型基因分型检测方法，通过多重探针阵列快速检测耐多药结核分枝杆菌分离株对利福平和异烟肼的耐药性。
设计了一种用于基因分型检测的多重探针阵列，以同时筛查结核分枝杆菌中与利福平和异烟肼耐药性相关的rpoB、katG、inhA和ahpC基因的突变，并添加了一个检测最近确认的异烟肼耐药性遗传标记之一ahpC - 6和 - 9位点的探针。通过使用所开发的基因分型检测方法，鉴定出52株耐多药分离株，其中46株rpoB基因有突变（88.5%），45株katG基因第315密码子、inhA调控区和oxyR - ahpC基因间区有突变（86.5%），而所有鉴定出的35株敏感分离株均显示野生型杂交模式。利福平耐药性的敏感性和特异性分别为88.5%和100%，异烟肼耐药性的敏感性和特异性分别为86.5%和100%。
通过所开发的多重探针阵列可实现对结核分枝杆菌分离株中rpoB、katG、inhA和ahpC基因突变引起的利福平和异烟肼耐药性的快速同时检测。
这种基因分型检测方案有潜力在临床应用中得到开发，用于在开始有效化疗前快速检测耐药结核分枝杆菌分离株。
