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[目的]初步探讨结核分枝杆菌散在分布重复单位(Mycobacterial interspersed repetitiveunits)和多位点串联重复序列(variablenumber of tandem repeats)技术基因分型技术在河南省结核病分枝杆菌基因分型中的应用,初步了解河南省局部地区MIRU-VNTR基因型种类及其分布。[方法]2007年在河南省的嵩县、新密、扶沟、中牟4个县及南阳市的结核病防治机构实验室分离培养的317株结核分枝杆菌,设计引物,应用PCR和琼脂糖凝胶电泳技术对结核分枝杆菌进行分型检测分析,基因分型聚类分析采用Bio-Numerics软件。[结果]对317株结核分枝杆菌临床分离株的12个MIRU位点检测结果显示明显的基因多态性。经基因聚类分析,可分5个基因群(Ⅰ群、Ⅱ群、Ⅲ群、Ⅳ群和Ⅴ群),292个基因型。其中Ⅰ群占4.7%,含15个基因型;Ⅱ群占67.8%,含198个基因型;Ⅲ群占22.7%,含64个基因型;Ⅳ群1.8%,含4个基因型;Ⅴ群3.5%,含11个基因型。[结论]初步表明河南省结核分枝杆菌MIRUs基因存在明显的多态性,至少存在5个MIRU型,主要流行型为Ⅱ型。
[Objective] To explore the application of Mycobacterial interspersed repetitive units and variable number of tandem repeats genotyping in Mycobacterium tuberculosis genotyping in Henan Province, To understand the genotype and distribution of MIRU-VNTR in some areas of Henan Province. [Method] A total of 317 strains of Mycobacterium tuberculosis were isolated and cultured in 2007 in Songxian, Xinmi, Fugou, Zhongmou and Nanyang TB control laboratories in Henan Province. Primers were designed for PCR and agarose coagulation Gel electrophoresis technology for genotyping detection and analysis of Mycobacterium tuberculosis, genotyping cluster analysis using Bio-Numerics software. [Results] The 12 MIRU loci of 317 clinical isolates of Mycobacterium tuberculosis showed obvious genetic polymorphisms. By gene cluster analysis, five gene clusters (Ⅰ group, Ⅱ group, Ⅲ group, Ⅳ group and Ⅴ group) and 292 genotypes were obtained. Among them, group Ⅰ accounted for 4.7% and contained 15 genotypes; group Ⅱ accounted for 67.8% and contained 198 genotypes; group Ⅲ accounted for 22.7% and contained 64 genotypes; group Ⅳ accounted for 1.8% and contained 4 genotypes; group Ⅴ 3.5%, containing 11 genotypes. [Conclusion] The preliminary results showed that there are obvious polymorphisms of MIRUs in Mycobacterium tuberculosis in Henan Province. There are at least five MIRU types and the main epidemic type is type.