结核分枝杆菌耐RFP快速检测的方法研究.pdfVIP

实用预防医学 2008年2月 第 l5卷 第1期 Practical Preventive Medicine，Feb．2008，Vol 15，No．1 217 文章编号：1006—3110(2008)O1—0217—02 结核分枝杆菌耐RFP快速检测的方法研究 张修明 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 rpoB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 采用PCR—SSCP检测痰标本和菌株中结核杆菌rpoB基因突变。 结果 50例耐RFP菌株中有45例PCR—SSCP条带与结核分枝杆菌H Rv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100％；12例敏感菌株与H Rv条带一致，35份肺结核病人痰标本，21份痰 PCR扩增 阳性，15份SSCP图谱与 H Rv图谱有差别，而且PCR—SSCP 28份图谱与H Rv有差别，阳性率为80％。 结论 PCR—SSCP检测结核分枝杆菌rpoB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，并将成 为直接检测l临床标本中结核分枝杆菌耐RFP快速方法。 关键词： 分枝杆菌，结核；RFP；聚合酶链反应；多态性，单链构象 中图分类号：R378．91 1 文献标识码：A Study on Rapid Detection of Rifampin——resistant Mycobacterium Tuberculosis by PCR and Single——strand Conformation Anal— ysis ZHANG xiU—ming Anxian County Center for Disease Control and Prevention，Anxian 415600，Hunan) Abstract： Obiective To evaluate rifampjn—resistant Mycobacterium tuberculosis by polymerase chain reaction—sin— gle strand conformation(PCR—SSCP)technique in detection of rpoB gene mutation of Mycobacterium tuberculosis in sputum specimens． Method rpoB gene mutation of Mycobacterium tuberculosis was detected with PCR—SSCP frOm clinical spu- lum specimens． Results The characteristics of PCR—SSCP band patterns in 50 rifampin—resistant strains were different frOm H17 Rv band patterns except 5 rifampin—resistant strains．1n comparison with susceptibility testing methods，the sensitivi． ty and specificity of PCR—SSCP were 90％ and 100％ respectively．PCR—SSCP band patterns of 12 sensitive strains were identica1 with those of H Rv．In 35 strains isolated frOm sputum specimens，2 1 were positive by PCR．The characteristics of 15 SSCP profiles and 28 PCR—SSCP profiles were both different frOm those of H ，Rv profiles，and the positive rate was 80％ ． Conclusion PCR—SSCP can easilY and rapi