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Comparative Genomic Hybridization Analysis of Enterococcus faecalis: Identification of Genes Absent from Food Strains

机译:粪肠球菌的比较基因组杂交分析:食品菌株缺乏基因的鉴定。

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摘要

Enterococcus faecalis, a member of the natural microbiota of animal and human intestinal tracts, is also present as a natural contaminant in a variety of fermented foods. Over the last decade, E. faecalis has emerged as a major cause of nosocomial infections. We investigated the genetic diversity in 30 clinical and food isolates, including strains V583 and MMH594, in order to determine whether clinical and food isolates could be distinguished. Data were obtained using comparative genomic hybridization and specific PCR with a total of 202 probes of E. faecalis, selected using the available V583 genome sequence and part of the MMH594 pathogenicity island. The cognate genes encoded mainly exported proteins. Hybridization data were analyzed by a two-component mixture model that estimates the probability of any given gene to be either present or absent in the strains. A total of 78 genes were found to be variable, as they were absent in at least one isolate. Most of the variable genes were clustered in regions that, in the published V583 sequence, related to prophages or mobile genetic elements. The variable genes were distributed in three main groups: (i) genes equally distributed between clinical and dairy food isolates, (ii) genes absent from dairy food-related isolates, and (iii) genes present in MMH594 and V583 strains only. Further analysis of the distribution of the last gene group in 70 other isolates confirmed that six of the probed genes were always absent in dairy food-related isolates, whereas they were detected in clinical and/or commensal isolates. Two of them corresponded to prophages that were not detected in the cognate isolates, thus possibly extending the number of genes absent from dairy food isolates. Genes specifically detected in clinical isolates may prove valuable for the development of new risk assessment markers for food safety studies and for identification of new factors that may contribute to host colonization or infection.
机译:粪肠球菌是动物和人肠道的天然微生物群的成员,在多种发酵食品中也作为天然污染物存在。在过去的十年中,粪肠球菌已经成为医院感染的主要原因。我们确定了30种临床和食品分离株的遗传多样性,包括菌株V583和MMH594,以确定是否可以区分临床和食品分离株。使用比较性的基因组杂交和特异性PCR,共202个粪肠球菌探针获得数据,使用可用的V583基因组序列和MMH594致病岛的一部分进行选择。同源基因主要编码输出蛋白。通过两组分混合模型分析杂交数据,该模型估计菌株中任何给定基因存在或不存在的概率。由于至少一个分离株中不存在78个基因,因此发现它们是可变的。大多数可变基因聚集在已发布的V583序列中与噬菌体或移动遗传元件有关的区域中。可变基因分为三个主要类别:(i)在临床和乳制品分离株之间平均分布的基因,(ii)与乳制品相关的分离株中不存在的基因,和(iii)仅存在于MMH594和V583菌株中的基因。进一步分析最后一个基因组在其他70个分离株中的分布,证实与乳制品相关的分离株中始终不存在所探测的基因中的6个,而在临床和/或常规分离株中都检测到了。它们中的两个对应于在同源分离物中未检测到的前噬菌体,因此可能扩大了乳制品分离物中不存在的基因的数量。在临床分离株中特异性检测到的基因对于开发用于食品安全性研究的新风险评估标志物以及鉴定可能有助于宿主定植或感染的新因素可能具有重要价值。

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