PCR-SSCP快速检测耐多药结核分枝杆菌  

Rapid detection of gene mutation in multiple-drug resistant mycobacterium tuberculosis by PCR and single-strand conformation polymorphism analysis

作　　者：宋薇薇[1] 华树成[2] 彭丽萍[2] 

机构地区：[1]中国人民解放军208医院呼吸内科,吉林长春130062 [2]吉林大学第一医院呼吸科,吉林长春130021

出　　处：《吉林医学》2006年第7期760-761,共2页Jilin Medical Journal

摘　　要：目的:了解本地区结核病耐药基因突变情况,探讨PCR-SSCP作为新的分子药敏试验方法在临床的应用价值。方法:通过提取耐INH、RFP、SM的肺结核患者痰中结核分枝杆菌DNA,进行PCR-SSCP分析,检测结核分枝杆菌rpoB、katG、rpsL基因是否存在突变,并与传统L-J药敏实验对照。结果:30株耐多药株中,耐RPF、INH、SM基因突变阳性率为90%(27/30)、63%(19/30)、53%(16/30)。3个基因联合突变共8株(26.7%),2个基因联合突变共18株(60%),即26株(86.7%)。单基因突变共2株,2株无基因突变。结论:通过PCR-SSCP方法可检测出绝大部分耐多药结核病的耐药基因,rpoB、katG、rpsL基因突变与本地区结核杆菌对RFP、INH、SM耐药性有关。与传统L-J药敏实验对比,PCR-SSCP是一种敏感、快速的指导临床用药的先进检测方法。Objective To establish PCR-SSCP for rapid analyzing resistant gene in multiple drug resistance of mycobacterium tuberculosis and to evaluate its use in clinics. Method 30 mycobacterium tuberculosis strains in multiple drug resistance were analyzed by PCR-SSCP and classic drug susceptibility tests. The fragments of rpoB, rpsL and katG were analyzed by single-stranded conformarion polymorphism （SSCP）. Results The gene mutation rate of rpoB, rpsL and katG resistance of isolated strains were 90% （27/ 30）,53% （16/30）,63%（19/30） Respectively. There are total 8（26.7%） strains with three genes mutation and 18（60%） strains with two genes mutation. Among the MDR-TB strains,there are 2 strains with single gene mutation and 2 strains without gene mutation, Conclusion Alterations in rpoB, rpsL, katG gene may be the important mechanisms of mycobacterium tuberculosis resistance to rifampin, streptomycin and isoniazid. The PCR-SSCP is an accurate and stable method to identify rpoB rpsL, katG showed mutation of mycobacterium tuberculosis, and it is useful for quickly identifying the MDR-TB strains in clinics.

关 键 词：聚合酶链反应 单链构象多态性 耐多药 结核分枝杆菌 突变基因 

分 类 号：R450[医药卫生—治疗学]