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首页> 外文期刊>Journal of Clinical Microbiology >Rapid Multiple-Locus Variant-Repeat Assay (MLVA) for Genotyping of Streptococcus agalactiae
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Rapid Multiple-Locus Variant-Repeat Assay (MLVA) for Genotyping of Streptococcus agalactiae

机译：无乳链球菌基因分型快速多位点变异重复测定（MLVA）

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Several methods have been used for typing of Streptococcus agalactiae (group B streptococci [GBS]). Methods currently in use may provide inadequate resolution (e.g., typing of capsular polysaccharides and surface protein) or are labor-intensive and expensive (e.g., multilocus sequence typing [MLST] or pulsed-field gel electrophoresis). This work describes the construction and use of a multiple-locus variant-repeat assay (MLVA) on 126 well-characterized human GBS strains, consisting mostly of invasive Norwegian strains and international reference strains. Based on in silico whole-genomic analysis of the genomes of strains A909, NEM316, and 2603V/R, 18 candidate loci were selected and investigated by PCR. Eleven loci showed diversity, and the five most diverse loci were used for the construction of an MLVA, consisting of a multiplex PCR followed by fragment analysis with capillary electrophoresis. The assay generated clusters which corresponded well with those observed by other methods. However, it provided a considerably higher degree of diversity, with 70 different MLVA types compared to 36 types generated by MLST. Simpson's index of diversity for the 5-locus MLVA was 0.963, compared to 0.899 for the MLST in this strain collection. MLVA results will generally be available within 2 days, which is usually faster than MLST. In our hands, MLVA of GBS represents a rapid, easy, and comparably inexpensive method for high-resolution genotyping of GBS.
机译：几种方法已被用于无乳链球菌（B群链球菌[GBS]）的分型。当前使用的方法可能提供不足的分辨率（例如，荚膜多糖和表面蛋白的分型）或者劳动强度大且昂贵（例如，多位点序列分型[MLST]或脉冲场凝胶电泳）。这项工作描述了对126个特征明确的人GBS菌株（主要由入侵性挪威菌株和国际参考菌株组成）的多位点变异重复测定（MLVA）的构建和使用。基于 in silico 的菌株A909，NEM316和2603V / R的基因组全基因组分析，选择了18个候选基因座并通过PCR进行了研究。 11个基因座显示出多样性，而5个多样性最高的基因座用于MLVA的构建，该基因座由多重PCR和随后的带有毛细管电泳的片段分析组成。该测定产生的簇与通过其他方法观察到的簇非常吻合。但是，与MLST生成的36种类型相比，它具有70种不同的MLVA类型，提供了更高的多样性。在该菌株收集中，五基因座MLVA的辛普森多样性指数为0.963，而MLST的辛普森多样性指数为0.899。 MLVA结果通常会在2天内获得，通常比MLST更快。在我们手中，GBS的MLVA代表了一种快速，简便且相对便宜的GBS高分辨率基因分型方法。

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