机构地区:[1]中国疾病预防控制中心传染病预防控制所传染病预防控制国家重点实验室,北京102206
出 处:《中华流行病学杂志》2009年第1期58-62,共5页Chinese Journal of Epidemiology
基 金:国家自然科学基金资助项目(30771853).
摘 要:目的评价结核分枝杆菌基因组中串联重复序列(VNTR)位点在中国结核分枝杆菌临床分离株的分型鉴定的分辨力。方法参考http://minisatellites.u-psud.fr/网站记载的位点选取其中VNTR位点,参照H37Rv结核分枝杆菌标准株全基因序列,利用DNAStar软件设计引物,采用PCR分别对中国结核分枝杆菌临床来分离株和H37Rv标准株的VNTR位点进行检测,根据PCR凝胶电泳图片中各菌株相应位点扩增产物的大小,确定重复单元的重复次数。计算Hunter-Gaston指数来分析各位点的分辨能力,及各位点合并分辨力。结果共检测135株中国结核分枝杆菌临床分离株和H37Rv标准株的45个VNTR位点。结果显示45个VNTR位点对136株菌的分辨力各不相同,Hunter-Gaston指数最大者为0.814(0.797-0.830),最小者为O.015(0.001-0.028),〉0.5的有13个位点。位点的合并分辨力分析显示,随着位点数量增加,分辨力增强,如Qubll-b和Qub18两个位点合并分析,Hunter-Gaston指数为0.936,分组数为44组;Qub11-b、Qub18、Mtub21、Rv2372、MIRU26、Qub26、Qub4156c、Qub11-a和Qub15等9个位点合并分析,Hunter-Gaston指数已经达到1,组数为136组,表示已达到最大分辨力,即株水平分型。结论不同VNTR位点具有不同的分辨力。其分型分辨力,多位点联合明显好于单位点。Qub11-b等9个位点的合并分型能力较好,这些位点有利于结核分枝杆菌的分型研究。Objective To evaluate the discriminatory efficiency of multiple loci of variable numbers of tandem repeats (VNTR) in Mycobaeterium tuberculosis genome. Genotyping and identification on Chinese M.tuberculosis clinical strains were used to locate a series of high discriminated loci, so as to provide the basis for creating a standardized multiple loci VNTR analysis (MLVA) to distribute fast typing and identification on Chinese M.tuberculosis. Methods VNTR loci which were chosen from the website http://minisatellites.u-psud.fr/ and referenced to the genome sequence of M.tuberculosis standard strain H37Rv were tested in Chinese M.tuberculosis clinical strains and H37Rv by means of PCR. The primers were designed by DNAStar software. The repeat member of VNTR unit was estimated by the result of PCR. The discrimination power of single locus or multiple loci was confirmed by the Hunter-Gaston index. Results 45 VNTR loci were tested in 135 Chinese M.tuberculosis clinical strains and H37Rv. The discrimination power of these loci appeared different from each other, with the biggest Hunter-Gaston index as 0.814 (0.797-0.830) , the smallest one as 0.015 (0.001-0.028) , and there were 13 loci with which the Hunter-Gaston index was bigger than 0.5. Results showed that the discrimination power was increasing by different loci that associated with each other. The more loci that were combined, the bigger the Hunter-Gaston index was. For example, the Hunter-Gaston index of Qub11-b associated with Qub18 was 0.936, by which 136 strains could be divided into 44 groups. With the combination of 9 loci including Qub 11 -b, Qub 18, Mtub21, Rv2372, MIRU26, Qub26, Qub4156c, Qub 11-a and Qubl 5, the Hunter-Gaston index could have reached 1 and by which the 136 strains could be divided into 136 groups, also showing that the biggest discrimination power to strain identification, viz. strain level genotype were reached. Conclusion The discrimination power of different locus was different. The discrimination power of multiple lo
关 键 词:结核分枝杆菌 可变数目串联重复序列 基闪分型
分 类 号:R378[医药卫生—病原生物学]
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