Variable-number-tandem repeat typing of mycobacterium tuberculosis isolates with low copy numbers of IS6110 by using mycobacterial interspersed repetitive units
Variable-number-tandem repeat typing of mycobacterium tuberculosis isolates with low copy numbers of IS6110 by using mycobacterial interspersed repetitive units
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DOI：
10.1128/jcm.40.5.1592-1602.2002
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发表时间：
2002-05-01
影响因子：
9.4
通讯作者：
Crawford, JT
中科院分区：
文献类型：
--
作者：
Cowan, LS;Mosher, L;Crawford, JT
使用最近引入的基于分枝杆菌散布重复单元的可变数目串联重复（MIRU-VNTR）的方法对具有低 IS6110 拷贝数的 180 个结核分枝杆菌和牛分枝杆菌分离株的研究集进行基因分型。将结果与更常用的方法 IS6110 限制性片段长度多态性 (RFLP) 和 spoligotyping 的结果进行比较。这些分离株是 1996 年至 1999 年在密歇根州收集的，作为该州新结核病例所有分离株基因分型项目的一部分。扩增了 12 个 MIRU 位点，并通过琼脂糖凝胶电泳分析扩增子以确定每个 MIRU 位点的拷贝数。 MIRU-VNTR 产生了比 IS6110 RFLP（58 个模式）更独特的模式（80 个模式），正如本研究组所预期的那样。 Spoligo 分型识别出 59 种模式。没有一种方法可以定义所有独特的分离株，所有三种分型方法的组合生成了 112 个不同的模式，识别了 90 个独特的分离株和 22 个簇中的 90 个分离株。结果证实了 MIRU-VNTR 分型的潜在效用，并表明需要使用多种方法进行分型才能获得最大特异性。
A study set of 180 Mycobacterium tuberculosis and Mycobacterium bovis isolates having low copy numbers of IS6110 were genotyped using the recently introduced method based on the variable-number tandem repeats of mycobacterial interspersed repetitive units (MIRU-VNTR). The results were compared with results of the more commonly used methods, IS6110 restriction fragment length polymorphism (RFLP) and spoligotyping. The isolates were collected in Michigan from 1996 to 1999 as part of a project to genotype all isolates from new cases of tuberculosis in the state. Twelve MIRU loci were amplified, and the amplicons were analyzed by agarose gel electrophoresis to determine the copy number at each MIRU locus. MIRU-VNTR produced more distinct patterns (80 patterns) than did IS6110 RFLP (58 patterns.), as would be expected in this study set. Spoligo-typing identified 59 patterns. No single method defined all unique isolates, and the combination of all three typing methods generated 112 distinct patterns identitying 90 unique isolates and 90 isolates in 22 clusters. The results confirm the potential utility of MIRU-VNTR typing and show, that typing with multiple methods is required to attain maximum specificity.