RAPD技术在唐山市结核分枝杆菌指纹分型研究中的应用  

THE APPLICATION OF RAPD TECHNOLOGY IN FINGERPRINTING OF MYCOBACTERIUM TUBERCULOSIS

作　　者：王东[1] 冯福民[1] 刘茜[1] 郭梅[1] 曹燕花[1] 李世明[2] 么作义 高宝霞[1] 孙永红[1] 

机构地区：[1]华北煤炭医学院预防医学系流行病与卫生统计学学科,唐山063000 [2]唐山市结核病医院

出　　处：《现代预防医学》2007年第17期3212-3214,共3页Modern Preventive Medicine

基　　金：卫生部科技攻关项目子课题(2003BA712A11-24)

摘　　要：[目的]RAPD DNA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RAPD DNA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]RAPD指纹图谱分析将102株分枝杆菌分为7种菌型,其中Ⅰ型35株(34.31%),Ⅱ型28株(27.45%),Ⅲ型24株(23.53%),Ⅳ型6株(5.88%),Ⅴ型3株(2.94%),Ⅵ型4株(3.92%),Ⅶ型2株(1.97%),Ⅰ、Ⅱ、Ⅲ型是主要的3种类型。RAPD扩增特征条带大小为201～1 018 bp。分组统计显示,3种主要的RAPD DNA指纹的分布与患者的居住地、职业、结核菌耐药性有关,而与患者的性别、年龄、结核接触史等无关。[结论]RAPD DNA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。[Objectives] To develop the RAPD molecular typing method and apply it in genotyping of Mycobacterium tuberculosis （MTB）. [Methods] 102 sputum specimens of tuberculosis patient were colleted and cultured at 37℃, the clinical isolates of MTB were obtained, and the phenotype and drug resistance were identified. The chromosomal DNA of MTB was extracted, and the characteristics of DNA fingerprints were analyzed with RAPD. [Results] The 102 strains of MTB were finally classified into 7 genotypes according to DNA fingerprinting. There were 35 strains of genotype Ⅰ （34.31%）, 28 strains of genotype Ⅱ （27.45%）, 24 strains of genotypeⅢ （23.53%）, 6 strains of genotypeⅣ （5.88%）, 3 strains of genotypeⅤ （2.94%）, 4 strains of genotypeⅥ （3.92%）, and 2 strains of genotypeⅦ （1.97%）, genotype Ⅰ , Ⅱ , Ⅲ were the main types. The specific segment amplified by RAPD was about 201-1 018 bp. The distribution of main genotypes had significant relation with inhabitancy, profession and drug resistance to MTB of TB patients, but had no relation with the gender, age, and history of TB contact of TB patients. [Conclusions] RAPD is a fast and simple molecular typing method for MTB genotyping, and the epidemic of three main MTB strains exists in Tangshan.

关 键 词：RAPD 结核分枝杆菌 基因分型 

分 类 号：Q789[生物学—分子生物学]