菌阴肺结核患者痰标本中结核分枝杆菌rpoB耐药基因突变的检测  

Detection of rpoB gene mutation of Mycobacterium tuberculosis in sputum negative specimens by smear and culture

作　　者：熊国亮[1] 张慧慧[1] 刘珍琼[1] 辛茶香[1] 张周云[1] 

机构地区：[1]江西省胸科医院检验科,南昌330006

出　　处：《江西医学检验》2006年第4期289-290,295,共3页Jiangxi Journal of Medical Laboratory Sciences

基　　金：江西省卫生厅科研资助(编号0301114)

摘　　要：目的应用套式PCR-DNA测序方法直接检测菌阴肺结核患者痰标本中结核分枝杆菌相关的rpoB基因突变,以期建立一种判定菌阴肺结核患者是否对利福平有无耐药性的方法。方法采用套式PCR-DNA测序方法直接检测112例活动性肺结核患者和20例非结核性肺部疾病患者痰标本中结核分枝杆菌rpoB基因突变情况。同份痰标本同时做涂片抗酸染色、罗氏培养。结果112例活动性肺结核患者痰标本39例为菌阴(涂阴培阴),39例菌阴痰中套式PCR扩增18例呈阳性,产物DNA测序2例有rpoB基因突变,位点为第526发生了CAC→GAC突变和第531发生了TCG→TTG突变。20例非结核性肺部疾病患者标本套式PCR扩增均为阴性,特异性100%。结论套式PCR-DNA测序可望为直接检测菌阴肺结核患者临床痰标本中结核分枝杆菌耐利福平的准确、特异、快速的方法。Objective To detect rpoB gene mutation of rifampin-resistant Mycobacterium tuberculosis in sputum negative specimens with smear and culture by nested PCR-DNA sequencing for developing a detective method of rifampin-resistant in Mycobaeterium tuberculosis. Methods 112 sputum specimens from the patients with active pulmonary tuberculosis and 20 sputum specimens from the patients with non-tuberculosis pulmonary disease were detected by nested PCR-DNA sequencing, smear and culture. Results 39 of 112 sputum specimens from the patients with active pulmonary tuberculosis were negative in.the smear and the culture, 2 of 39 sputum negative specimens by smear and culture had rpoB mutation, with＇the identical mutations（CAC→GAC）at codon 526 of the rpoB gene, and （TCG→TTG） at codon 531 of the rpoB gene. No positive ones was found in 20 sputum specimens from the non-tuberculosis pulmonary diseases. Conclusion Nested PCR-DNA sequencing may become an accuracy, specific, rapid method for directing the detection of rpoB gene mutation in Mycobacterium tuberculosis.

关 键 词：分枝杆菌 结核 利福平 抗药性 微生物 

分 类 号：R446.5[医药卫生—诊断学] R378.911[医药卫生—临床医学]