VNTR技术用于安徽省耐药结核分支杆菌基因分型的初步研究  

Genotyping of drug-resistant Mycobacterium tuberculosis strains isolated from the patients in Anhui province with variable number tandem repeat method

作　　者：董海燕[1] 王庆[2] 刘志广[1] 赵秀芹[1] 万康林[1] 

机构地区：[1]中国疾病预防控制中心传染病预防控制所,北京102206 [2]安徽省肺科医院

出　　处：《中华微生物学和免疫学杂志》2005年第8期687-690,共4页Chinese Journal of Microbiology and Immunology

基　　金：国家"十五"重点项目资助(项目号2001DEA10074);国家自然科学基金项目(项目号30471526)

摘　　要：目的探讨VNTR(variable number of tandem repeat)技术在安徽省耐药结核分支杆菌基因分型中的应用。方法初步选取13个分型效果较好的VNTR基因位点。应用聚合酶链反应(PCR)和琼脂糖凝胶电泳,建立检测耐药结核分支杆菌DNA指纹多态性的方法,分析耐药结核分支杆菌DNA多态性。结果共对78株耐药结核分支杆菌的13个VNTR位点进行了检测,根据这些菌株的指纹多态性特征,共分为4个基因型(Ⅰ型、Ⅱ型、Ⅲ型、Ⅳ型),分别为Ⅰ型3.8%(378)、Ⅱ型6.4%(578)、Ⅲ型10.3%(878)、Ⅳ型所占比例最大,为79.5%(6278)。在Ⅳ型菌中,耐药菌株主要为耐多药(结核分支杆菌至少耐异烟肼和利福平两种药)和单耐利福平菌株,其所占比例分别为45.8%和22.6%。结论本资料分析表明,安徽省耐药结核分支杆菌的传播似以Ⅳ型菌株为主,应加强此型菌株流行的监控。Objective To genotype resistant Mycobacterium tuberculosis strains isolated from the patients in Anhui province with a new genotyping method based on the variable-number tandem repeats（VNTR）. Methods To select 13 VNTR loci and develop a fast fingerprinting technology for M. tuberculosis strains by polymerase chain reaction（PCR） and agarose gel electrophoresis technique and analyzed the polymorphism of DNA fingerprinting of drug-resistant M. tuberculosis. Results Seventy-eight M. tuberculosis strains detected with 13 VNTR loci were classified into four types（Ⅰ,Ⅱ,Ⅲ,Ⅳ） based on their characters of PCR amplified fingerprints, type Ⅰ 3.8% （3/78）,type Ⅱ 6.4%（5/78）, type Ⅲ 10.3%（8/78） and type Ⅳ 79.5%（62/78）. In type Ⅳ group, the main drug-resistant strains were multidrug-resistant ones （45.8%） and rifampin-resistant strains（22.6%）. Conclusion The results showed that the prevalence of drug-resistant tuberculosis was primarily due to the transmission of type Ⅳ in Anhui orovince. It is suggested that surveillance of genotype Ⅳ needs to be strengthened.

关 键 词：结核分支杆菌 VNTR技术 耐药 基因分型 耐药结核分支杆菌 VNTR位点 安徽省 聚合酶链反应(PCR) 步研究 DNA多态性 

分 类 号：R378[医药卫生—病原生物学]