间隔区寡核苷酸分型技术用于结核分枝杆菌多重感染的初步研究  

Multiple infections of Mycobacterium tuberculosis with spoligotyping

作　　者：吴小翠[1,2] 王晓樱[3] 魏剑浩 刘海灿[2,4] 赵秀芹[2,4] 夏远志[1,2] 楼永良[1] 吕建新[1] 万康林[1,2,4] 

机构地区：[1]温州医科大学检验医学院生命科学学院,温州325035 [2]中国疾病预防控制中心传染病预防控制所,传染病预防控制国家重点实验室,北京102206 [3]四川大学华西基础医学与法医学院病理生理教研室,成都610041 [4]感染性疾病诊治协同创新中心,杭州310003

出　　处：《中国人兽共患病学报》2015年第1期1-5,共5页Chinese Journal of Zoonoses

基　　金：国家科技重大专项课题(No.2013ZX10003002-001)资助~~

摘　　要：目的初步评价间隔区寡核苷酸分型(Spoligotyping)技术在鉴定结核分枝杆菌多重感染中的应用。方法选取结核分枝杆菌临床分离株,5μm孔径滤膜过滤法制备单个细菌菌悬液,平板接种37℃培养,选取生长良好的单个菌落培养增菌,提取基因组DNA,Spoligotyping进行基因分型鉴定,比较临床分离株与相应的单克隆分离株的分型结果。结果共选取32株结核分枝杆菌临床分离株,获得306个单克隆分离株。Spoligotyping分型鉴定结果显示30株结核分枝杆菌临床分离株与相应的单克隆分离株的分型结果一致,2株临床分离株的单克隆分离株呈现了多种基因型,多重感染率为6.25%。结论 Spoligotyping可用于快速,有效地区分结核分枝杆菌多重感染。We assessed the application of spacer oligotyping（Spoligotyping）for identifying the multiple infections of Mycobacterium tuberculosis（M.tuberculosis）preliminarily in this study.Clinical M.tuberculosis isolates were randomly selected and prepared to single colony suspensions by filtration method with 5μm pore diameter membranes.Then those single colony suspensions were inoculated on Lwenstein-Jensen slants at 37℃,from which the cultures of all M.tuberculosis colonies were gathered and genomic DNA samples were extracted.The original clinical isolates and corresponding monoclonal strains were typed by means of Spoligotyping to compare the genotypes.Results showed that 32 clinical M.tuberculosis isolates and 306 corresponding monoclonal strains were obtained.And the results of spoligotyping showed that 286 monoclonal strains had same spoligotypes as 30 original clinical isolates,but the monoclonal strains from two original clinical isolates showed diversity.The multiple infections rate of M.tuberculosis was 6.25%.It＇s suggested that spoligotyping can be used to distinguish multiple infections of M.tuberculosis quickly and effectively.

关 键 词：结核分枝杆菌 间隔区寡核苷酸分型 多重感染 

分 类 号：R378.91[医药卫生—病原生物学]