淮南矿区煤工尘肺结核患者耐多药结核分枝杆菌耐药基因rpoB序列分析  

Sequence analysis on drug-resistant gene of rpoB in MDR-TB among pneumoconiosis patients complicated with tuberculosis in Huainan mining district

作　　者：陆军[1] 叶松[1] 李朝品[2] 秦蕊[1] 李卫鹏[3] 

机构地区：[1]安徽理工大学医学院,淮南232001 [2]安徽皖南医学院,淮南232001 [3]蚌埠医学院附属医院

出　　处：《中华劳动卫生职业病杂志》2012年第8期579-581,共3页Chinese Journal of Industrial Hygiene and Occupational Diseases

基　　金：2010年安徽省高校省级重点自然科学研究项目（KJ2010A087）,2011年淮南市科技计划项目（2011A07902）

摘　　要：目的分析淮南矿区尘肺结核患者感染的耐多药结核分枝杆菌（multiple drugs resistant bacillus tuberculosis，MDR．TB）利福平耐药基因rpoB突变特点。方法收集结核分枝杆菌临床分离株114株，采用药敏试验鉴定MDR．TB，抽提MDR．TBDNA和H37Rv标准菌株DNA，PCR法扩增rpoB基因，并应用全自动DNA测序仪对rpoB基因的突变集中区域进行测序分析。结果从114株结核分枝杆菌临床分离株中检出MDR^B31株（27．19％，31／114），31株MDR．TB中rpoB基因突变率为93．55％（29／31），主要集中在531（45．16％，14／31）和526位点（29．03％，9／31）碱基置换突变，其次是516（3．23％，1／31）、535位点（3．23％，1／31）；1株526位和531位同时突变，1株511位和526位同时突变，2株未发现rpoB基因突变。随机检测的lO株利福平敏感株未见rpoB基因单链构象异常。结论高度保守的rpoB基因突变是导致尘肺结核患者MDR．TB利福平耐药的分子基础，其突变位点呈多样性。Objective To study the characteristics of drug-resistant genetic mutation of rpoB in multiple drugs resistant bacillus tuberculosis （MDR-TB） among patients of pneumoconiosis complicated with pulmonary tuberculosis. Methods A total of 114 clinical isolated strains of Mycobacterium tuberculosis were collected, MDR-TB were identified by conventional antimicrobial susceptibility test （AST）. Their genomes DNA were extracted, the target genes were amplified by PCR, and the hot regions in the rpoB gene were analyzed by automated DNA sequenator. Results The results by AST showed that there were 31 strains of MDR-TB in the 114 clinical isolated strains, the rate of drug resistance was 27.19% （31/114）. No mutation of rpoB was identified in 10 rifampicin-sensitive strains that randomly chosen, while conformation changes were found in MDR-TB strains, and the mutation rate of rpoB was 93.55% （29/31） in resistant strains, mainly concentrated in codon 531 （45.16%, 14/31 ） and 526 （29.03%, 9/31）, happened base substitutions, including 27 unit point mutation and 2 two point mutation. Conclusions The substitution of highly conserved amino acids encoded by rpoB gene results in the molecular mechanism responsible for RFP resistance in MDR-TB among patients of pneumoconiosis complicated with pulmonary tuberculosis. It also proves that moB gene is diversiform.

关 键 词：尘肺 结核 MDR-TB 耐药 RPOB基因 序列分析 

分 类 号：R135.2[医药卫生—劳动卫生] R521[医药卫生—公共卫生与预防医学]