Palma Federica, Pasquali Frédérique, Lucchi Alex, De Cesare Alessandra, Manfreda Gerardo
Department of Food and Agricultural Sciences, Alma Mater Studiorum- University of Bologna, Bologna, Italy.
Ital J Food Saf. 2017 Sep 28;6(3):6879. doi: 10.4081/ijfs.2017.6879. eCollection 2017 Aug 16.
is a food-borne pathogen able to survive and grow in different environments including food processing plants where it can persist for month or years. In the present study the discriminatory power of Whole Genome Sequencing (WGS)-based analysis (cgMLST) was compared to that of molecular typing methods on 34 isolates collected over one year in the same rabbit meat processing plant and belonging to three genotypes (ST14, ST121, ST224). Each genotype included isolates indistinguishable by standard molecular typing methods. The virulence potential of all isolates was assessed by Multi Virulence-Locus Sequence Typing (MVLST) and the investigation of a representative database of virulence determinant genes. The whole genome of each isolate was sequenced on a MiSeq platform. The cgMLST, MVLST, and identification of virulence genes were performed using publicly available tools. Draft genomes included a number of contigs ranging from 13 to 28 and N50 ranging from 456298 to 580604. The coverage ranged from 41 to 187X. The cgMLST showed a significantly superior discriminatory power only in comparison to ribotyping, nevertheless it allows the detection of two singletons belonging to ST14 that were not observed by other molecular methods. All ST14 isolates belonged to VT107, which 7-loci concatenated sequence differs for only 4 nucleotides to VT1 (Epidemic clone III). Analysis of virulence genes showed the presence of a fulllength version in all ST14 isolates and of a mutated version including a premature stop codon (PMSC) associated to attenuated virulence in all ST121 isolates.
是一种食源性病原体,能够在包括食品加工厂在内的不同环境中存活和生长,在食品加工厂中它可以持续存在数月或数年。在本研究中,将基于全基因组测序(WGS)的分析(cgMLST)的鉴别能力与分子分型方法对在同一兔肉加工厂一年内收集的34株分离株的鉴别能力进行了比较,这些分离株属于三种基因型(ST14、ST121、ST224)。每种基因型包括通过标准分子分型方法无法区分的分离株。通过多毒力基因座序列分型(MVLST)和对毒力决定基因代表性数据库的研究,评估了所有分离株的毒力潜力。在MiSeq平台上对每个分离株的全基因组进行测序。使用公开可用的工具进行cgMLST、MVLST和毒力基因鉴定。草图基因组包含13至28个重叠群,N50范围为456298至580604。覆盖范围为41至187X。cgMLST仅与核糖体分型相比显示出显著更高的鉴别能力,然而它能够检测到属于ST14的两个单株,这是其他分子方法未观察到的。所有ST14分离株都属于VT107,其7个基因座串联序列与VT1(流行克隆III)仅相差4个核苷酸。毒力基因分析表明,所有ST14分离株中存在全长版本,而所有ST121分离株中存在包括与减弱毒力相关的提前终止密码子(PMSC)的突变版本。