摘要
目的探讨淮南矿区尘肺并发结核患者感染耐多药结核分枝杆菌(multiple drugs resistant Mycobacterium tuberculosis,MDR-MTB)L型异烟肼耐药基因katG突变特点。方法收集114株MTB菌L型临床分离株,用药敏试验鉴定MDR-MTB菌L型;抽提MDR-MTB L型和H37Rv标准菌株DNA,PCR法扩增katG基因,并对katG基因的突变集中区域进行测序分析。结果从114株临床分离株中检出MDR-MTB菌L型31株(27.2%),其katG基因突变率为61.3%(19/31);主要集中在315位点碱基置换突变,以Ser315Thr为主(AGC→ACC)(48.4%,15/31),其次是315位点Ser315Asn(AGC→AAC)突变(9.7%,3/31)以及431位点Ala431Val(GCG→GTG)突变(3.2%,1/31),未发现多位点联合突变,12株(38.7%,12/31)未发现katG基因突变;随机检测的10株异烟肼敏感株未见katG基因单链构象异常。结论淮南矿区尘肺并发结核患者感染的MDR-MTB菌L型异烟肼耐药情况较为严重,高度保守的katG基因突变是导致异烟肼耐药的分子基础,其突变位点呈多样性。
Objective To investigate the mutational characteristics of isoniazid-resistant gene katG in multiple drugs resistant Mycobacterium tuberculosis （MDR-MTB） L-form from the patients with pneumoconiosis and tuberculosis in Huainan mining district. Methods A total of 114 clinical isolates of MTB L-form were collected, and MDR-MTB were identified by the conventional antimicrobial susceptibility test （AST）. The genome DNA from MDR-MTB L-form and standard strain H37Rv was extracted, and the katG gene was amplified by PCR. Then, the hot mutation regions in katG gene were analyzed by direct sequencing. Results Thirty-one （27.2%） MDR-MTB L-form strains were detected from 114 clinical isolates, and the mutation rate of katG gene was 61.3% （ 19/31 ）. The mutation sites were mainly located on codon 315, including Ser315Thr （AGC→ACC, 15/31,48.4% ） and Ser315Asn （AGC→AAC, 3/ 31, 9.7% ） , and codon 431 （Ala431Val, GCG→GTG, 1/31, 3.2% ）. No multisite mutation was found. Moreover, no single strand conformation abnormity of katG gene was found in randomly collected 10 isoniazid-sensitive strains. Conclusion There was serious isoniazid-resistance of MDR-MTB L-form in the patients with pneumoconiosis and tuberculosis in Huainan mining district. The mutation of highly conserved katG gene was responsible for isoniazid-resistance of MDR-MTB L-form, and there were diversiform mutation sites.
出处
《临床检验杂志》
CAS
CSCD
北大核心
2013年第6期434-437,共4页
Chinese Journal of Clinical Laboratory Science
基金
国家自然科学基金(81172778)
安徽省高校省级重点自然科学研究项目(KJ2010A087
KJ2012A081)
2011年淮南市科技计划项目(2011A07902)
2011年安徽理工大学中青年学术骨干基金
关键词
尘肺
结核
耐多药结核分枝杆菌L型
耐药
KATG基因
序列分析
pneumoconiosis
tuberculosis
multiple drugs resistant Mycobacterium tuberculosis L-form
drug-resistance
katG gene
sequence analysis
作者简介
陆军，1972年生，男，副教授，硕士研究生导师，博士研究生，主要从事病原生物学检验、环境污染与人类健康等研究。
通信作者：李朝品，教授，博士研究生导师，E-mail：epli001@126．com。
