结核杆菌利福平耐药与rpoB基因突变的相关性研究
Correlation between rifampin-resistance and rpoB gene mutation in Mycobacterium tuberculosis
卢阳 1薛欣 1楚雍烈1
作者信息
- 1. 西安交通大学医学院病原生物学及免疫学系,陕西,西安,710061
- 折叠
摘要
目的:分析西安地区耐利福平(RIF)结核杆菌临床分离株rpoB基因突变的特点,探讨快速检测结核杆菌耐RIF基因型的方法.方法:对120株西安地区收集的结核杆菌临床分离株(药敏结果为耐RIF 81株,对RIF敏感39株),分别采用聚合酶链反应.单链构象多态性(PCR-SSCP)和等位基因特异性多重聚合酶链反应(MAS-PCR)检测rpoB基因突变情况.结果:PCR-SSCP检测,耐RIF菌株中rpoB基因突变发生率为85%(69/81),39株敏感株中仅有1株发生突变,突变率为2%(1/39).MAS-PCR检测,耐RIF菌株中rpoB基因突变发生率为93%(75/81),39株敏感株中未发现rpoB基因突变;531,516和526位点突变的比例分别为45%(34/75),24%(18/75)和20%(15/75).结论:西安地区结核杆菌临床分离株耐RIF与rpoB基因突变相关,MAS-PCR检测rpoB基因可作为临床耐RIF结核杆菌的简单快速准确的早期诊断方法.
Abstract
AIM: To analyze the character of rpoB genemutation in rifampin (RIF)-resistant Mycobacterium tuberculosis (M. TB) strains isolated from tuberculosis patients in Xi'an and to develop and evaluate methods to detect the RIF-resistant gene of M. TB. METHODS: RpoB gene mutation was examined in 120 M. TB strains (81 RIF-resistant and 39 RIF-sensitive) isolated from tuberculosis patients in Xi'an by polymerase chain reactionsingle strand conformation polymorphism (PCR-SSCP) and multiplex allele specific polymerase chain reaction (MAS-PCR). RESULTS: By PCR-SSCP, the ratio of rpoB mutation in RIF-resistant M. TB strains was 85% (69/81) and there was only 1strain with rpoB mutation in the 39 RIF-sensitive M.TB strains. By MAS-PCR, the ratio of rpoB gene mutation in RIF-resistantM. TB strains was 93% (75/81) and no rpoB gene mutation was found in the 39 RIF-sensitive M. TB strains. The ratio of rpoB gene mutation point at codon 531, codon 516 and codon 526 was respectively 45% ( 34/75 ), 24% (18/75) and 20% ( 15/75 ). CONCLUSION: RIF resistance of M. TB strains isolated from tuberculosis patients in Xi'an is correlative with rpoB gene mutation. Detection of rpoB gene using MAS-PCR can be a simple, rapid and accurate method for the early diagnosis of RIF-resistant M.TB.
关键词
结核杆菌/rpoB基因/利福平/等位基因特异性多重聚合酶链反应Key words
mycobacterium tuberculosis/rpoB gene/rifampin/multiplex allele specific-polymerase chain reaction引用本文复制引用
基金项目
西安市科技支撑计划基金(YF075158)
出版年
2009
NETL
NSTL
维普
万方数据