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首页> 外文期刊>Journal of Clinical Microbiology >Discriminatory Power and Reproducibility of Novel DNA Typing Methods for Mycobacterium tuberculosis Complex Strains
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Discriminatory Power and Reproducibility of Novel DNA Typing Methods for Mycobacterium tuberculosis Complex Strains

机译：结核分枝杆菌复杂菌株新型DNA分型方法的鉴别力和可重复性

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In recent years various novel DNA typing methods have been developed which are faster and easier to perform than the current internationally standardized IS6110 restriction fragment length polymorphism typing method. However, there has been no overview of the utility of these novel typing methods, and it is largely unknown how they compare to previously published methods. In this study, the discriminative power and reproducibility of nine recently described PCR-based typing methods for Mycobacterium tuberculosis were investigated using the strain collection of the interlaboratory study of Kremer et?al. (J. Clin. Microbiol. 37:2607-2618, 1999). This strain collection contains 90 M. tuberculosis complex and 10 non-M. tuberculosis complex mycobacterial strains, as well as 31 duplicated DNA samples to assess reproducibility. The highest reproducibility was found with variable numbers of tandem repeat typing using mycobacterial interspersed repetitive units (MIRU VNTR) and fast ligation-mediated PCR (FLiP), followed by second-generation spoligotyping, ligation-mediated PCR (LM-PCR), VNTR typing using five repeat loci identified at the Queens University of Belfast (QUB VNTR), and the Amadio speciation PCR. Poor reproducibility was associated with fluorescent amplified fragment length polymorphism typing, which was performed in three different laboratories. The methods were ordered from highest discrimination to lowest by the Hunter-Gaston discriminative index as follows: QUB VNTR typing, MIRU VNTR typing, FLiP, LM-PCR, and spoligotyping. We conclude that both VNTR typing methods and FLiP typing are rapid, highly reliable, and discriminative epidemiological typing methods for M. tuberculosis and that VNTR typing is the epidemiological typing method of choice for the near future.
机译：近年来，已经开发出各种新颖的DNA分型方法，它们比当前国际标准化的IS 6110 限制性片段长度多态性分型方法更快，更容易执行。但是，还没有关于这些新颖的打字方法的实用性的概述，并且在很大程度上如何将它们与以前发表的方法进行比较尚不清楚。在这项研究中，使用Kremer等人的实验室间研究的菌株收集品，研究了九种最近描述的基于PCR的结核分枝杆菌的判别能力和可重复性。 （J.Clin.Microbiol.37：2607-2618，1999）。该菌株收集物包含90M。结核病复合物和10个非 M。结核病复杂的分枝杆菌菌株以及31个重复的DNA样品，以评估可重复性。使用分枝杆菌散布的重复单位（MIRU VNTR）和快速连接介导的PCR（FLiP）进行可变数目的串联重复键入，然后进行第二代spoligotyping，连接介导的PCR（LM-PCR）和VNTR分型，发现了最高的重复性使用在贝尔法斯特女王大学（QUB VNTR）鉴定的5个重复基因座和Amadio物种PCR。可重复性差与在三个不同实验室中进行的荧光扩增片段长度多态性分型有关。按照Hunter-Gaston判别索引，从最高辨别力到最低辨别力，将方法按以下顺序排序：QUB VNTR分型，MIRU VNTR分型，FLiP，LM-PCR和Spoligotyping。我们得出结论，VNTR分型方法和FLiP分型方法都是针对 M的快速，高度可靠和有区别的流行病学分型方法。结核病和VNTR分型是近期流行的流行病分型方法。

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