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首页> 外文期刊>Applied Microbiology >High-Throughput Typing Method To Identify a Non-Outbreak-Involved Legionella pneumophila Strain Colonizing the Entire Water Supply System in the Town of Rennes, France
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High-Throughput Typing Method To Identify a Non-Outbreak-Involved Legionella pneumophila Strain Colonizing the Entire Water Supply System in the Town of Rennes, France

机译：高通量分型方法，用于鉴定在法国雷恩镇定居整个供水系统的非暴发性嗜肺军团菌菌株

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Two legionellosis outbreaks occurred in the city of Rennes, France, during the past decade, requiring in-depth monitoring of Legionella pneumophila in the water network and the cooling towers in the city. In order to characterize the resulting large collection of isolates, an automated low-cost typing method was developed. The multiplex capillary-based variable-number tandem repeat (VNTR) (multiple-locus VNTR analysis [MLVA]) assay requiring only one PCR amplification per isolate ensures a high level of discrimination and reduces hands-on and time requirements. In less than 2 days and using one 4-capillary apparatus, 217 environmental isolates collected between 2000 and 2009 and 5 clinical isolates obtained during outbreaks in 2000 and 2006 in Rennes were analyzed, and 15 different genotypes were identified. A large cluster of isolates with closely related genotypes and representing 77% of the population was composed exclusively of environmental isolates extracted from hot water supply systems. It was not responsible for the known Rennes epidemic cases, although strains showing a similar MLVA profile have regularly been involved in European outbreaks. The clinical isolates in Rennes had the same genotype as isolates contaminating a mall's cooling tower. This study further demonstrates that unknown environmental or genetic factors contribute to the pathogenicity of some strains. This work illustrates the potential of the high-throughput MLVA typing method to investigate the origin of legionellosis cases by allowing the systematic typing of any new isolate and inclusion of data in shared databases.
机译：在过去的十年中，法国雷恩市发生了两次军团菌病暴发，需要深入监测供水网络中的嗜肺军团菌和该市的冷却塔。为了表征所得的大量分离株，开发了一种自动化的低成本分型方法。基于多重毛细管的可变数目串联重复序列（VNTR）（多位点VNTR分析[MLVA]）测定法每个分离株仅需进行一次PCR扩增，可确保较高的区分度并减少动手和时间要求。在不到2天的时间内，使用一台4毛细管仪器，分析了2000年至2009年之间收集的217种环境分离株以及2000年至2006年雷恩暴发期间获得的5种临床分离株，并鉴定了15种不同的基因型。具有密切相关基因型且代表77％人口的一大群分离株完全由从热水供应系统提取的环境分离株组成。尽管已知相似的MLVA病毒株经常参与欧洲爆发，但它对已知的雷恩流行病不负责任。雷恩的临床分离株与污染购物中心冷却塔的分离株具有相同的基因型。这项研究进一步证明未知的环境或遗传因素会导致某些菌株的致病性。这项工作通过允许对任何新分离株进行系统分型并将数据包括在共享数据库中，说明了高通量MLVA分型方法调查军团菌病病例起源的潜力。

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