快速检测结核分枝杆菌γpoB基因突变的研究  

Rapid technique in detection γpoB gene mutation in mycobacterium tuberculosis by PCR-SSCP

作　　者：刘国华[1] 闫国蕊[2] 刘传玉[2] 祖燕[2] 贾振 王子玲[2] 李英 

机构地区：[1]河南省卫生防疫站,河南郑州450003 [2]河南省结核病防治研究所,河南郑州450003 [3]周口市结核病防治所,河南周口466000 [4]焦作市结核病防治所,河南焦作454152

出　　处：《河南医学研究》2001年第4期316-318,共3页Henan Medical Research

基　　金：河南省科技攻关项目 ( 98110 70 40 8)

摘　　要：目的 :应用PCR SSCP技术直接快速检测痰标本中结核分枝杆菌γpoB基因突变 ,评价此方法用于结核分枝杆菌利福平 (RFP)耐药性的意义。方法 :以结核分枝杆菌H37Rv为对照、5 0例耐RFP(单耐和多耐 )结核分枝杆菌临床分离株、12例RFP敏感菌株、35例耐RFP肺结核患者痰标本 ,采用PCR SSCP检测痰标本和菌株中结核杆菌γpoB基因突变。结果 :5 0例耐RFP菌株中有 45例PCR SSCP条带与结核分枝杆菌H37Rv条带有明显差别 ,与药敏试验结果做对比 ,PCR SSCP检测敏感性为 90 % ,特异性为 10 0 % ;12例敏感菌株与H37Rv条带一致 ,35份肺结核病人痰标本 ,2 1份痰PCR扩增阳性 ,15份SSCP图谱与H37Rv图谱有差别 ,而用PCR SSCP 2 8份图谱与H37Rv有差别 ,阳性率为 80 %。结论 :PCR SSCP检测结核分枝杆菌γpoB基因突变快速、简便、易行 ,有望用于结核分枝杆菌RFP耐药性的快速测定 。Objective: To evaluate rifampin resistant Mycobacterium tuberculosis by PCR SSCP technique to detect γpoB gene mutation in mycobacterium tuberculosis of sputums.Methods: Fifty clinicae isolates of rifampin resistant and multidruy rdsistant mycobacterium tuberculosis,twelve sensitive strains, thirty five rifampin resistant strains from sputum specimens of patients with pulmonary tuberculosis, were analyzed by PCR SSCP technique, with H37Rv reference as control group.Results: The characteristics of PCR SSCP graph in fifty strains of rifampin resistant were different from H37Rv except for five strains.In comparison with susceptibility testing methods, the sensitivity and specificity were 90% and 100% respectively.Twelve sensitive strains were same as H37Rv.In thirty five strain from sputum spcimens of patients with pulmonary tuberculosis, twenty one strains of them were positive by PCR.The characteristic of SSCP graph in fifteen of them were different from H37Rv,and the characteristics of PCR SSCPgraph in twenty eight of them were different from H37Rv,the positivity was 80%.Conclusion: The PCR SSCP can easily and rapidly detect rifampin resistance of mycobacterium tuberculosis,the technigue might be used for rapidly detect mycobacterium tuberculosis rifampin resistance in clinical specimen directly.

关 键 词：结核分枝杆菌 γpoB基因 聚合酶链反应 多态现象 单链构象 

分 类 号：R521[医药卫生—内科学]