多种基因分型技术在识别结核分枝杆菌基因型的联合应用  

The combined application of multiple genotyping methods in identifying genotypes of Mycobacterium tuberculosis strain circulating in rural China

作　　者：胡屹[1] 蒋伟利[1] 赵琦[1] 王伟炳[1] 徐飚[1] 

机构地区：[1]复旦大学公共卫生学院流行病学教研组,上海200032

出　　处：《中华结核和呼吸杂志》2009年第8期576-580,共5页Chinese Journal of Tuberculosis and Respiratory Diseases

基　　金：国家自然科学基金资助项目（30771843）;上海重点学科建设项目资助项目（B118）

摘　　要：目的评价MTB散在分布重复单位（MIRUs）和IS5110限制性内切酶片段长度的多态性（IS6110-RFLP）基因分型技术，以及两者联合应用在我国华东农村地区MTB分子流行病学研究中的应用及其效果。方法以人群为基础的横断面研究设计，采用比例法确定2004-2005年从浙江省德清县和江苏省灌云县收集的351株MTB菌株的耐药性，利用间隔区寡核苷酸分型方法识别北京基因型MTB，并进行MIRUs和IS6110-RFLP独立基因分型和串联联合应用。结果先对351株MTB菌株采用MIRUs基因分型可以识别235种基因型，包括46个成簇基因型和189个惟一基因型，分辨力为0．9317；用IS6110-RFLP可进一步从成簇基因型中识别出28个亚组，分辨力为0．9989。相对而言，先对目标菌株采用IS6110一RFLP基因分型，可识别267基因型，分辨力为0．9684，包括46个成簇基因型和221个惟一基因型，再用MIRUs基因分型技术可进一步从成簇基因型中识别出31个亚型，分辨力为0．9992。两种顺序不同联合分型方法的基因分型的辨别能力相似，尤其是在耐多药MTB（分辨力分别为0．9965和0．9963）及北京基因型MTB（分辨力分别为0．9930和0．9933）的分型中。结论MIRUs分型的方法简便、快速，结果有效、可靠，在大规模的结核病分子流行病学研究中，可先采用MIRUs，再对成簇MTB菌株进行IS6110-RFLP二次分型的联合分型策略。Objective To evaluate the performance of MIRUs （mycobacterial interspersed repetitive units） genotyping alone, IS6110-RFLP （IS6110 restriction fragment length polymorphism） genotyping alone and their combination applied in the molecular epidemiological study of Mycobacterium tuberculosis （ MTB ） isolates circulating in rural China. Methods A cross-sectional study was designed to collect MTB isolates from the TB patients registered in local TB dispensaries of Deqing county and Guanyun county from 2004 to 2005. The proportion method was used to determine drug susceptibility of MTB isolates to the first line anti-TB drugs （isoniazid, rifampin, ethambutol and streptomycin）. The Beijing family of MTB was identified by Spoligotyping. All isolates were genotyped by MIRUs alone, IS6110-RFLP alone and their combination. Results Of the 351 studied MTB isolates, 243 （69. 2% ） had the genotypes that belonged to the Beijing family, and 223 （63.5%） were resistant to at least 1 anti-TB drug, including 53 （15. 1% ） resistant to isoniazid and rifampin simultaneously or multidrug resistant （MDR）. The heterogeneity of 12 MIRUs loci differed from 0. 76 in MIRU26 to 0. 003 in MIRU2. MIRUs alone identified 235 genotgpes （HGI =0. 9317）,including 46 ＂clusters＂ containing 162 isolates and 189 ＂unique＂ pattern/isolates. Thirty-eight isolates comprised the largest MIRUs defined cluster and presented MIRUs type 2233 2517 3533. MIRUs based clusters were further identified by IS6110-RFLP into 28 subgroups containing 80 isolates （HGI = 0. 9989）. In comparison, IS6110-RFLP determined 267 genotypes from all isolates, including 46 ＂clusters＂ containing 130 isolates （HGI =0. 9684） and 221 ＂unique＂ pattern/isolates. All the 46 IS6110-RFLP defined clusters could also be further identified by MIRUs into 31 subgroups （HGI =0. 9992）. The performance of MIRUs prior to IS6110-RFLP combination was comparable to that of IS6110-RFLP prior to MIRUs combination, especially in Beijing genotype str

关 键 词：分枝杆菌 结核 串联重复序列 限制性内切酶图谱法 基因型 

分 类 号：R686[医药卫生—骨科学]