DNA指纹技术检测耐多药结核分枝杆菌的应用研究

摘要/Abstract

摘要： 目的应用结核分枝杆菌DNA指纹技术,了解耐多药结核分枝杆菌的基因型状况并探讨耐多药结核病的分子流行病学。方法以结核分枝杆菌插入序列IS6110序列为模板,设计一对特异外向引物,建立一种结核分枝杆菌DNA指纹技术方法,对临床标本中分离的220株耐多药结核分枝杆菌进行DNA指纹基因分析。同时对实验数据进行聚类分析,进行耐多药结核病分子流行病学研究。结果根据结核分枝杆菌DNA指纹图谱分析,220株耐多药结核分枝杆菌均产生特征DNA指纹图谱,DNA指纹图谱变异很大。每一菌株DNA指纹的拷贝数介于3至18之间。其中大部分菌株,即205株（93.2%）包含613个拷贝,平均为11个带。这205株DNA指纹图谱中,162株为特异的,提示它们在流行病学上是独立的。有58株（26.4%）指纹图谱组成了15个簇,每簇28株,它们的IDNA指纹图谱相同,可能代表了近期的传播。尤其是在此DNA指纹图谱中,其中分子量为200 bp扩增带频率最大,在220株耐多药结核分枝杆菌均出现。结论结核杆菌DNA指纹技术检测可应用耐多药结核分枝杆菌DNA指纹图谱分型,并可用于耐多药结核病分子流行病学调查。

关键词: 抗药性,多药, 分枝杆菌,结核, 印迹法,DNA

Abstract: Objective To investigate genotypes of mutidrug-resistant(MDR) M.tuberculosis strains By DNA fingerprinting technique,and to discuss their role in molecular epidemiology.Methods A pair of outer primers for the amplification of IS6110 sequence from M.tuberculosis was designed.DNA fingerprinting method was established and used to analyze the genotypes of 220 MDR-resistant M.tuberculosis clinical isolates.The data from all of the tests were performed cluster analysis,and molecular epidemiology analysis.Results According to analysis of the DNA fingerprinting patterns,220 strains of Mycobacterium tuberculosis isolated were all shown specific DNA fingerprinting patterns,which were highly variable.The number of DNA copies per strain varied from 3 to 18.The majority(205,93.2%) of the strains contained 6-13 copies,with a mean of 11 bands.Of 205 DNA fingerprinting patterns observed,162 were unique,indicating epidemiological independence.However,58(26.4%) strains clustered in 15 groups,each cluster consisted of 2-8 isolates with identical patterns,presumably representing cases of recent transmission.In particular,one distinct pattern within this DNA fingerprint group was shared,220 strains of M.tuberculosis isolated all found one 200bp molecular weight of pattern.Conclusion DNA fingerprinting technique might be effectively applicable as a tool for the detection of genotype of drug-resistant M.tuberculosis,and for the survey on molecular epidemiology of multidrug resistance tuberculosis(MDR-TB).

Key words: Drug resistance,multiple, Mycobacterium tuberculosis, Blotting,southern

周琳,谭守勇,唐林国,梁志强,刘志辉,陈其琛,蔡杏珊,黎意芬,李妙琼,. DNA指纹技术检测耐多药结核分枝杆菌的应用研究[J]. 中国防痨杂志, 2007, 29(6): 502-507.

Zhou Lin,Tan Shouyong,Tang Linguo,et al.. Detection of mutidrug-resistant Mycobacterium tuberculosis using DNA fingerprinting technique[J]. Chinese Journal of Antituberculosis, 2007, 29(6): 502-507.

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