MLVA用于新疆部分地区结核分枝杆菌基因分型的初步研究

目的探讨结核菌株进行多位点可变数目串联重复序列分析(the multiple locus VNTR analysis,MLVA)。方法选择2010年至2011年新疆维吾尔自治区第五次结核病流行病学抽样调查资料,采用经典24位点方法进行基因分型。并采用Bio Numerics5.0数据库进行基因聚类分析。将结核分枝杆菌原始株,取一菌环溶于400μl TE中悬菌,80℃1 h灭活,12 000 r/min离心10 min,弃上清,600μl TE重新悬菌,进行多位点串联重复序列分析。结果新疆99株结核分枝杆菌分为2个基因群:分别为基因群Ⅰ和基因群Ⅱ,基因群Ⅰ66株(66.7%),基因株Ⅱ33株(33.3%);基因群Ⅰ是北京家族,基因群Ⅰ的66株结核分枝杆菌有65种不同的基因型,有2株结核分枝杆菌属于同一簇,成簇率为1.5%,基因群II33株结核分枝杆菌菌株的MLVA图谱不同,成簇率为0。结论新疆结核分枝杆菌菌株存在明显的基因多态性,以北京基因型菌株为主,同时还存在一定比例的非北京基因型,应加强对主要流行菌株流行的监控及管理。

Preliminary study on genotype of Mycobacterium tuberculosis in some areas of Xinjiang by the multiple locus VNTR analysis

Objective To explore the multiple locus VNTR analysis (MLVA) of Mycobacterium tuberculosis.Methods It was selected that the data of the fifth TB epidemiological sampling srvvey in Xinjiang Uygur Autonomously Region from 2010 to 2011,and the genotyping was carried out by the classic 24 site method.The genetic clustering was analyzed by the BioNumerics5.0 database.The original strains of Mycobacterium tuberculosis,the annulus were dissolved and suspended bacterium in 400 μl TE,80 ℃ 1 h inactivated,12 000 r/min,the centrifugal 10 min,abandon supernatant,suspended bacterium in 600 μl TE again;multiple site tandem repeat sequence analysis.Results There were two genetic groups in Xinjiang:gene group Ⅰ and gene group][Ⅱ,66 strains in genetic group Ⅰ (66.7％),and 33 strains in gene strain Ⅱ (33.3％).gene growp Ⅰ was Beijing genotype strains 65 different genotype from gene group of 66 strains of Mycobacterium tuberculosis Ⅰ,2 strains of Mycobacterium tuberculosis belong to the same cluster,clusters rate was 1.5％,It was different that the gene group of Ⅱ 33 of Mycobacterium tuberculosis strain of MLVA pictures,clusters rate was 0.Conclusion Mycobacterium tuberculosis strains exist obvious gene polymorphism in Xinjiang,with the majority of Beijing genotype strains,and a certain proportion of strains are not Beijing genotype,we should strengthen the monitoring and management for major epidemic strains.