套式PCR在检测结核分枝杆菌embB基因突变中的应用价值

The Value of Nested PCR in Detecting EmbB Gene Mutations of Mycobacterium Tuberculosis

目的采用套式PCR(二次扩增)替代传统PCR(一次扩增)扩增痰中结核分枝杆菌embB基因,探讨其在检测结核分枝杆菌embB基因突变中的应用价值。方法选取本院已确诊的活动性肺结核住院病人68例和非结核肺部感染病人16例,嘱其留取晨痰,分别进行套式PCR扩增和传统PCR扩增,再行产物分析。结果68例活动性肺结核病人传统PCR扩增embB基因阳性结果为29例,阳性率为42.6%,套式PCR扩增embB基因阳性结果为47例,阳性率为69.1%,两者比较差异有显著性(χ2=9.66,P<0.05);16例非结核病人传统PCR和套式PCR和检测结果均为阴性,两者特异度均为100%。结论检测结核分枝杆菌embB基因突变,套式PCR灵敏度高于传统PCR,两者特异度相同,套式PCR能快速、敏感、特异地扩增结核分枝杆菌embB基因片段。

Objective To detect embB gene mutation of ethambutol-resistance Mycobacterium tuberculosis by nested PCR, and to evaluate their clinical value. Methods Sijxty-ejight sputum specimens from the patients with active pulmonary tuberculosis and 16 sputum specimens from the patients with non-tuberculous pulmonary disease were detected by nested PCR and normal PCR. Results Twenty-nine of 68 sputum specimens from the patients with active pulmonary tubercolosis by normal PCR were positive, and the positive rate was 42.6%; 47 of 68 sputum specimens by nested PCR were positive, and the positive rate was 69. 1%. No positive result was found in 16 sputum specimens from the non-uberenlosis pulmonary disease by nested PCR and normal PCR. Conclusion Nested PCR is better than normal PCR in detecting embB gene mutation of ethambutol-resistance Mycobacterium tuberculosis. Nested PCR may become for directing detection of embB gene mutation in Mycobacterium tuberculosis.