套式PCR及测序法检测痰中结核分枝杆菌耐乙胺丁醇embB基因突变  

Detection of EmbB Gene Mutation of Ethambatol-resistant in Mycobaterium Tuberculosis from Sputum Specimens by Nested PCR and DNA Sequencing

作　　者：刘珍琼[1] 刘金辉[2] 熊国亮[1] 辛茶香[1] 

机构地区：[1]江西省胸科医院检验科 [2]南昌大学医学院微生物研究室,南昌330006

出　　处：《江西医学院学报》2008年第4期22-24,共3页Acta Academiae Medicinae Jiangxi

基　　金：江西省卫生厅课题(050314)

摘　　要：目的应用套式PCR及测序法直接检测痰中结核分枝杆菌相关的embB基因突变,以期建立一种直接、快速、准确地检测结核分枝杆菌耐乙胺丁醇的实验方法。方法采用套式PCR扩增技术及测序法直接检测130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embB基因突变情况,同份痰标本同时做涂片抗酸染色、罗氏培养及菌型鉴定。结果29例耐乙胺丁醇标本有18例发生基因突变,均为306位密码子突变,基因突变率为62.1%(18/29)。结论套式PCR及测序法可望成为一种快速、特异、准确地检测痰中结核分枝杆菌embB基因突变的好方法。Objective Todetect embB gene mutation of ethambatol-resistant in sputum specimens by nested PCR and DNA sequencing,and expected to develop a direct, rapid and accuracy method for the detection of ethambatol-resistant in Mycobaterium tuberculosis. Methods One hundred and thirty sputum specimens from the patients with active pulmonary tuberculosis and 20 sputum specimens from the patients with non-tuberculous pulmonary disease were detected by nested PCR and DNA sequencing, with smear, culture and species identification at the same time. Results Eighteen of 29 sputum specimens with ethambatol-resistant strains had embB mutations with all mutations at codon 306, and the mutation rate was 62.1% （18/29）. Conclusion Nested PCR-DNA sequencing may become a rapid, specific, accuracy method for direct defection of embB gene mutation in Mycobacterium tuberculosis from clinical specimens.

关 键 词：套式PCR 分枝杆菌 结核 乙胺丁醇 DNA测序 基因 

分 类 号：R446.1[医药卫生—诊断学] R378.911[医药卫生—临床医学]