河南省结核分枝杆菌间隔区寡核苷酸分型技术的应用研究

Evaluation of the Spacers Oligonucleotide typing as a practical approach in molecular epidemiology of Mycobacterium tuberculosis in Henan

目的研究间隔区寡核苷酸序列(Spoligotyping)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,了解河南省基因型种类和分布以及北京家族基因型在河南省的分布特征。方法应用2007年在河南省结核病防治机构实验室分离培养的310株结核分枝杆菌,设计引物,应用PCR和间隔区寡核苷酸分型(Spoligotyping)技术对结核分枝杆菌进行分型检测分析,基因分型分析经http://www.miru-vntrplus.org/MIRU/index.faces网站分析,统计学数据经卡方检验。结果对310株结核分枝杆菌临床分离株的Spoligotyping结果显示分为4个基因群(Beijing和Beijing近似群、T群、Manu群、S群),24个基因型,有267株为北京家族基因型,占86.1%,且北京家族基因型与MDR耐药有相关性;河南省较年轻人群(<60岁)感染的结核分枝杆菌中,北京家族基因型所占比例高于年老组(即>60岁组),北京家族基因型与年轻组的联系强于年老组;北京家族基因型的分布在不同的地区间有差异。结论河南省结核分枝杆菌以北京家族基因型为主要流行型,且与耐药及年龄、地域有相关性,重点监测北京家族基因型。

Objective To Evaluation genotyping method Spoligotyping for Mycobacterium tuberculosis,Understand the types and distribution of genotypes of Mycobacterium tuberculosis clinical isolates and distribution features of Beijing family genotype in henan province.Methods M.tuberculosis clinical isolates selected from Henan province randomly were cultured with Lowenstein-Jensen medium.The bacteria from which the genome DNA was extracted were collected.The clustering of genotype was analyzed on http：//www.miru-vntrplus.org/MIRU/index.faces website.Chi-Square test was used for statistical analysis.Results The Spoligotyping pattern of 310 M.tuberculosis clinical isolates were analyzed respectively.The clustering of genotype showed that these strains could be categorized into 4 gene clusters（Beijing and Beijing like、T、Manu、S）and 24 genotypes.267 strains were Beijing family genotype accounting for 86.1 %,and shows the association between the Beijing family and the MDR-TB.The Beijing family genotype proportion in younger groups is higher than the older groups.The association between young age and Beijing strains was even stronger in younger ages.There are different characteristics of Beijing family genotype isolated in different regions.Conclusions The popular Genotype of the M.tuberculosis in Henan is Beijing family,The association between Beijing family and drug resistance、age、geography is significant.We should strengthen monitoring the Beijing family genotype.