13个VNTR位点用于113株结核分支杆菌的基因分型研究

曹晓慧; 蒋毅; 张媛媛; 刘志广; 赵秀芹; 金秀琴; 韩宝龙; 徐瑞兴; 刘敬华; 吕晶; 薛小洛; 万康林

Abstract

曹晓慧,蒋毅,张媛媛,刘志广,赵秀芹,金秀琴,韩宝龙,徐瑞兴,刘敬华,吕晶,薛小洛,万康林.13个VNTR位点用于113株结核分支杆菌的基因分型研究[J].Chinese journal of Epidemiology,2006,27(8):705-708

13个VNTR位点用于113株结核分支杆菌的基因分型研究

Study on the genotyping of 113 Mycobacterium tuberculosis strains isolated in Beijing based on 13 variable number of tandem DNA repeats

Received:January 26, 2006

DOI：

KeyWord: 结核分支杆菌基因分型聚类分析数目可变串联重复序列技术

English Key Word: Mycobacterium tuberculosis Genotyping Cluster analysis Variable number of tandem repeats

FundProject:国家自然科学基金资助项目(30471526)

Author Name	Affiliation	E-mail
CAO Xiao-hui	Institute for Infectious Disease Control and Prevention, Chinese Center for Disease Control and Prevention, Beijing 102206, China
JIANG Yi	Institute for Infectious Disease Control and Prevention, Chinese Center for Disease Control and Prevention, Beijing 102206, China
ZHANG Yuan-yuan	Institute for Infectious Disease Control and Prevention, Chinese Center for Disease Control and Prevention, Beijing 102206, China
LIU Zhi-guang	Institute for Infectious Disease Control and Prevention, Chinese Center for Disease Control and Prevention, Beijing 102206, China
ZHAO Xiu-qin	Institute for Infectious Disease Control and Prevention, Chinese Center for Disease Control and Prevention, Beijing 102206, China
JIN Xiu-qin	北京市第二医院 100031
HAN Bao-long	北京市第二医院 100031
XU Rui-xing	北京市第二医院 100031
LIU Jing-hua	Institute for Infectious Disease Control and Prevention, Chinese Center for Disease Control and Prevention, Beijing 102206, China
LV Jing	北京市第二医院 100031
XUE Xiao-luo	北京市第二医院 100031
WAN Kang-lin	Institute for Infectious Disease Control and Prevention, Chinese Center for Disease Control and Prevention, Beijing 102206, China	wankanglin@icdc.cn

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Abstract:

目的应用数目可变串联重复序列(VNTR)分子分型技术,对北京地区113株肺结核临床分离菌株进行分型研究,探讨北京地区菌株DNA多态性和基因型特征。方法采用PCR和琼脂糖凝胶电泳技术对结核分支杆菌13个VNTR位点进行检测,并应用Gel-Pro analyzer 3．1软件和BioNumerics 3．0软件进行结果分析。结果113株结核分支杆菌可分为4个基因型(分别为Ⅰ、Ⅱ、Ⅳ和Ⅴ型),其中Ⅰ型占92．0％(104／113),其他3型所占比例很小,分别为Ⅱ型占4．4％(5／113),Ⅳ和Ⅴ型均为1．8％(2／113),而标准菌株H37Rv在分型中为独立的一个基因型,即Ⅲ型。结论北京地区的结核分支杆菌存在基因多态性,其主要流行型为Ⅰ型。

English Abstract:

Objective Variable Number of Tandem Repeats(VNTRs) analysis was a recently developed method which could serve as a 'real-time' genotyping tool for Mycobacterium tuberculosis.One hundred and thirteen M. tuberculosis isolates from the patients with tuberculosis in Beijing were analysed using the reference method to study the characters of genetic diversity and genotype.Methods Thirteen tandem repeat loci(ETR-A,ETR-C,ETR-D,MIRU10，MIRU16, MIRU27，MIRU31，MIRU40，Mtub21， Mtub30, Mtub38, Qubl l a, Qubl l b) in the total genome of MTB were analyzed by PCR and agarose gel electrophoresis method. The characters of the polymorphism of DNA fingerprinting of one hundred and thirteen MTB strains were analyzed with Gel-Pro analyzer 3.1 software and BioNumerics 3.0 software.Results One hundred and thirteen MTB strains were characterized and classified in to four genotype families(type I，type 1I，type N，type v)based on thirteen tandem repeat loci. One hundred and four isolates( 92. 0 96)belonged to type 1，the other three genotypes scattered, five strains(4.4 0,6)remaining with type 11，while type N and type V having the same quantity 1.80.6 (2/113).M. tuberculosis H37Rv belonged to a unattached genotype( type ).Conclusion There was obvious length间ymorphism in the M.tuberculosis isolates which implied that type j was the epidemic strain clusters in M. tuberculosis in Beijing. VNTRs analysis seemed to be a simple, rapid, sensitive and valuable tool for epidemiological studies of M.tuberculosis complex organisms.

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