DRE-PCR分型方法在唐山市结核分枝杆菌研究中的应用

目的DRE-PCRDNA指纹分型方法的建立及其在分枝杆菌基因分型中的应用。方法收集结核病患者痰标本,分离培养获得分枝杆菌,进行表型鉴定及耐药性检测;提取基因组DNA,在建立DRE-PCRDNA指纹分型方法的基础上对分枝杆菌进行DNA指纹图谱分析。结果DRE-PCR指纹图谱分析将80株分枝杆菌分为13种菌型,其中A型菌26株(32.5%),B型菌14株(17.5%),C型菌16株(20.0%),其余各型均<7.5%。分组统计显示,3种主要的DRE-PCRDNA指纹在40~60岁组的分布偏高(P=0.019),而与患者的性别、职业、长期居住地、结核接触史、痰涂片、耐药性等无关。结论DRE-PCRDNA指纹分型方法是一种简单、快速的分枝杆菌的分型方法;3种主要类型的结核分枝杆菌在唐山地区流行。

Application of the double repetitive element-PCR (DRE-PCR) based DNA fingerprinting analysis to the genotyping of Mycobacterium tuberculosis

To establish the double repetitive element-PCR (DRE-PCR) molecular typing method to be used in the genotyping of Mycobacterium tuberculosis(MTB), 80 sputum specimens of patients with pulmonary tuberculosis were collected and cultivated at 37℃ to obtain the clinical isolates of MTB. These isolates were identified by means of the phenotypic method. And their chromosomal DNA was extracted. The DNA-fingerprinting analysis of MTB was performed on the basis of the established method of DRE-PCR. As analyzed with this method, the 80 MTB isolates were finally classified into 13 genotypes, in which most of them belonged to A (32.5%), B (17.5%) and C (20.0%) genotypes and the other 10 genotypes appeared to be scattered. There was a significant difference among different age groups in the DRE-PCR fingerprinting.This result indicates that the DRE-PCR based DNA fingerprinting analysis is proved to be a fast and simple molecular typing method used for the genotyping, and there exists three main genotypes in the epidemic strains of Mycobacterium tuberculosis. in Tangshan city.