Furukawa Maika, Goji Noriko, Janzen Timothy W, Thomas Matthew C, Ogunremi Dele, Blais Burton, Misawa Naoaki, Amoako Kingsley K
a Canadian Food Inspection Agency (CFIA) National Centres for Animal Disease, Lethbridge Laboratory, P.O. Box 640, Township Road 9-1, Lethbridge, AB T1J 3Z4, Canada.
b Laboratory of Veterinary Public Health, Department of Veterinary Medical Science, Faculty of Agriculture, University of Miyazaki, 1-1 Gakuenkibanadai-nishi, Miyazaki, 889-2192, Japan.
Can J Microbiol. 2018 Jan;64(1):75-86. doi: 10.1139/cjm-2017-0496. Epub 2017 Oct 31.
Serotyping of Salmonella enterica subsp. enterica is a critical step for foodborne salmonellosis investigation. To identify Salmonella enterica subsp. enterica serovars, we have developed a new assay based on a triplex polymerase chain reaction (PCR) with pyrosequencing for amplicon confirmation and phylogenetic discrimination of strains. The top 54 most prevalent serovars of S. enterica in Canada were examined with a total of 23 single-nucleotide polymorphisms (SNPs) and (or) single-nucleotide variations (SNVs) located on 3 genes (fliD, sopE2, and spaO). Seven of the most common serovars, Newport, Typhi, Javiana, Infantis, Thompson, Heidelberg, and Enteritidis, were successfully distinguished from the other serovars based on their unique SNP-SNV combinations. The remaining serovars, including Typhimurium, ssp I:4,[5],12:i:-, and Saintpaul, were further divided into 47 subgroups that demonstrate the relatedness to phylogenetic classifications of each serovar. This pyrosequencing assay is not only cost-effective, rapid, and user-friendly, but also provides phylogenetic information by analyzing 23 selected SNPs. With the added layer of confidence in the PCR results and the accuracy and speed of pyrosequencing, this novel method would benefit the food industry and provides a tool for rapid outbreak investigation through quick detection and identification of common S. enterica serovars in Canada.
肠炎沙门氏菌亚种的血清分型是食源性沙门氏菌病调查的关键步骤。为了鉴定肠炎沙门氏菌亚种血清型，我们开发了一种基于三重聚合酶链反应（PCR）并结合焦磷酸测序的新方法，用于扩增子确认和菌株的系统发育鉴别。我们利用位于3个基因（fliD、sopE2和spaO）上的总共23个单核苷酸多态性（SNP）和（或）单核苷酸变异（SNV），对加拿大最常见的54种肠炎沙门氏菌血清型进行了检测。根据其独特的SNP - SNV组合，成功区分出了7种最常见的血清型，即纽波特、伤寒、贾维亚纳、婴儿、汤普森、海德堡和肠炎血清型。其余血清型，包括鼠伤寒、ssp I:4,[5],12:i: -和圣保罗血清型，进一步分为47个亚组，这些亚组显示了各血清型与系统发育分类的相关性。这种焦磷酸测序方法不仅具有成本效益、快速且用户友好，还通过分析23个选定的SNP提供系统发育信息。由于PCR结果的可信度增加以及焦磷酸测序的准确性和速度，这种新方法将使食品行业受益，并为通过快速检测和鉴定加拿大常见的肠炎沙门氏菌血清型进行快速疫情调查提供一种工具。
