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利用新型微阵列对肠炎沙门氏菌进行毒力特性分析:在非典型宿主鼠伤寒沙门氏菌中检测和评估细胞致死性扩张毒素基因活性

Virulence Characterization of Salmonella enterica by a New Microarray: Detection and Evaluation of the Cytolethal Distending Toxin Gene Activity in the Unusual Host S. Typhimurium.

作者信息

Figueiredo Rui, Card Roderick, Nunes Carla, AbuOun Manal, Bagnall Mary C, Nunez Javier, Mendonça Nuno, Anjum Muna F, da Silva Gabriela Jorge

机构信息

Faculty of Pharmacy and Center for Neuroscience and Cell Biology, University of Coimbra, Coimbra, Portugal; Department of Bacteriology, Animal and Plant Health Agency, Weybridge, New Haw, Addlestone, Surrey, KT15 3NB, United Kingdom.

Department of Bacteriology, Animal and Plant Health Agency, Weybridge, New Haw, Addlestone, Surrey, KT15 3NB, United Kingdom.

出版信息

PLoS One. 2015 Aug 5;10(8):e0135010. doi: 10.1371/journal.pone.0135010. eCollection 2015.

Abstract

Salmonella enterica is a zoonotic foodborne pathogen that causes acute gastroenteritis in humans. We assessed the virulence potential of one-hundred and six Salmonella strains isolated from food animals and products. A high through-put virulence genes microarray demonstrated Salmonella Pathogenicity Islands (SPI) and adherence genes were highly conserved, while prophages and virulence plasmid genes were variably present. Isolates were grouped by serotype, and virulence plasmids separated S. Typhimurium in two clusters. Atypical microarray results lead to whole genome sequencing (WGS) of S. Infantis Sal147, which identified deletion of thirty-eight SPI-1 genes. Sal147 was unable to invade HeLa cells and showed reduced mortality in Galleria mellonella infection model, in comparison to a SPI-1 harbouring S. Infantis. Microarray and WGS of S. Typhimurium Sal199, established for the first time in S. Typhimurium presence of cdtB and other Typhi-related genes. Characterization of Sal199 showed cdtB genes were upstream of transposase IS911, and co-expressed with other Typhi-related genes. Cell cycle arrest, cytoplasmic distension, and nuclear enlargement were detected in HeLa cells infected by Sal199, but not with S. Typhimurium LT2. Increased mortality of Galleria was detected on infection with Sal199 compared to LT2. Thus, Salmonella isolates were rapidly characterized using a high through-put microarray; helping to identify unusual virulence features which were corroborated by further characterisation. This work demonstrates that the use of suitable screening methods for Salmonella virulence can help assess the potential risk associated with certain Salmonella to humans. Incorporation of such methodology into surveillance could help reduce the risk of emergence of epidemic Salmonella strains.

摘要

肠炎沙门氏菌是一种人畜共患的食源性病原体,可导致人类急性肠胃炎。我们评估了从食用动物及其产品中分离出的106株沙门氏菌菌株的毒力潜力。高通量毒力基因微阵列显示,沙门氏菌致病岛(SPI)和黏附基因高度保守,而前噬菌体和毒力质粒基因则存在差异。分离株按血清型分组,毒力质粒将鼠伤寒沙门氏菌分为两个簇。非典型微阵列结果导致对婴儿沙门氏菌Sal147进行全基因组测序(WGS),该测序鉴定出38个SPI-1基因缺失。与携带SPI-1的婴儿沙门氏菌相比,Sal147无法侵袭HeLa细胞,并且在大蜡螟感染模型中显示出较低的死亡率。首次在鼠伤寒沙门氏菌中确定了鼠伤寒沙门氏菌Sal199的微阵列和WGS,其存在cdtB和其他与伤寒相关的基因。Sal199的特征表明,cdtB基因位于转座酶IS911的上游,并与其他与伤寒相关的基因共表达。在感染Sal199的HeLa细胞中检测到细胞周期停滞、细胞质扩张和核增大,但在感染鼠伤寒沙门氏菌LT2的细胞中未检测到。与LT2相比,感染Sal199后检测到大蜡螟的死亡率增加。因此,使用高通量微阵列可快速鉴定沙门氏菌分离株;有助于识别异常的毒力特征,并通过进一步表征得到证实。这项工作表明,使用合适的沙门氏菌毒力筛选方法有助于评估某些沙门氏菌对人类的潜在风险。将这种方法纳入监测可以帮助降低流行沙门氏菌菌株出现的风险。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/aca8/4526557/b7e9097ea37f/pone.0135010.g001.jpg

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