基因型分析法快速检测耐药结核分枝杆菌  

Rapid genotype assay for detection of drug-resistant Mycobacterium tuberculosis

作　　者：石瑞如[1] 高飞絮[1] 沈青玉[1] 黄祖峰 秦殊[1] 王伟[1] 张国龙[1] 

机构地区：[1]河南省胸科医院检验科中美结核病研究中心,郑州450008

出　　处：《中华生物医学工程杂志》2014年第1期46-50,共5页Chinese Journal of Biomedical Engineering

基　　金：河南省医学科技攻关计划重大项目（201001014）

摘　　要：目的 探讨基因型分析法检测结核分枝杆菌异烟肼和利福平耐药性的价值.方法 采用多重PCR和线性探针反向膜杂交法来检测异烟肼耐药基因katG S315T和inhA C-15T突变以及利福平耐药基因rpoB D516V,H526Y,H526D,S531L突变来判断78株结核分枝杆菌临床分离株的耐药性,并与金标准L-J固体培养基药敏法以及BACTEC960液体培养药敏法进行对比分析.结果 基因型分析法在6h内可以完成;结核分枝杆菌常规药敏检测需要3个月.与后者相比,基因型分析法检测异烟肼耐药的敏感性和特异性分别为89％ （16/18）和99％ （77/78）;检测利福平耐药的敏感性和特异性均为100％（13/13,78/78）.结论 基因型分析法检测结核菌耐药性快速准确,对耐药结核病的早期诊断和治疗很有帮助,有望在临床实验室广泛开展.Objective To explore the value of the genotype assays detection of the resistance of Mycobacterium tuberculosis to isoniazid and rifampin.Methods Seventy-eight clinical isolates susceptible of harboring Mycobacterium tuberculosis were subject to the mutation assay of katG gene S315T,inhA gene C-15Tand rpoB genes D516V,H526Y,H526D,S531L by using multiplex polymerase chain reaction-based linear probe membrane hybridization for analysis of isoniazid and rifampin resistance.These results were compared with the L-J solid medium culture,the gold standard,and MGIT BACTEC 960 drug sensitivity test.Results Genotype assay was associated with shorter time for the detection compared with routine drug sensitivity test （6 hours vs 3 months）.The genotype assay yielded the sensitivity and specificity of 89％ （16/18） and 99％ （77/78） for isoniazid and 100％ and 100％ （13/13 and 78/78） for rifampin.Conclusion Genotype assay of Mycobacterium tuberculosis offers a rapid and accurate approach for the early diagnosis and treatment of drug-resistant tuberculosis,rendering it a promising technique to be extensively applied in clinical laboratories.

关 键 词：分支杆菌 结核 微生物敏感性试验 广泛耐药结核 线性探针反向膜杂交法 基因型 

分 类 号：R440[医药卫生—诊断学]