Department of Medical Microbiology, School of Medicine, University of Manchester, Clinical Sciences Building, Manchester Royal Infirmary, Oxford Road, Manchester M13 9WL, United Kingdom.
J Clin Microbiol. 2010 Apr;48(4):1055-60. doi: 10.1128/JCM.01566-09. Epub 2010 Feb 17.
Food-borne salmonellosis is a major manifestation of gastrointestinal disease in humans across the globe. Accurate and rapid identification methods could positively impact the identification of isolates, enhance outbreak investigation, and aid infection control. The SNaPshot multiplex system is a primer extension-based method that enables multiplexing of single nucleotide polymorphisms (SNPs). Here the method has been developed for the identification of five Salmonella serotypes, commonly detected in the United Kingdom, based on serotype-specific SNPs identified in the multilocus sequence typing (MLST) database of Salmonella enterica. The SNPs, in genes hemD, thrA, purE, and sucA, acted as surrogate markers for S. enterica serovars Typhimurium, Enteritidis, Virchow, Infantis, and Braenderup. The multiplex primer extension assay (MPEA) was conducted in two separate panels and evaluated using 152 Salmonella enterica isolates that were characterized by MLST. The MPEA was shown to be 100% specific and sensitive, within this collection of isolates. The MPEA is a sensitive and specific method for the identification and detection of Salmonella serotypes based upon SNPs seen in MLST data. The method can be applied in less than 6 h and has the potential to improve patient care and source tracing. The utility of the assay for identification of Salmonella serotypes directly from clinical specimens and food samples warrants further investigation.
食源性沙门氏菌病是全球人类胃肠道疾病的主要表现。准确快速的鉴定方法可以积极影响分离物的鉴定，增强暴发调查，并有助于感染控制。SNaPshot 多重系统是一种基于引物延伸的方法，可实现单核苷酸多态性 (SNP) 的多重化。在这里，该方法是基于沙门氏菌 enterica 的多位点序列分型 (MLST) 数据库中鉴定的血清型特异性 SNP，开发用于鉴定在英国常见的五种沙门氏菌血清型。hemD、thrA、purE 和 sucA 基因中的 SNP 作为 S. enterica 血清型 Typhimurium、Enteritidis、Virchow、Infantis 和 Braenderup 的替代标记。多重引物延伸分析 (MPEA) 在两个单独的面板中进行，并使用通过 MLST 表征的 152 株沙门氏菌 enterica 分离株进行评估。该 MPEA 在该分离株集合中显示出 100%的特异性和敏感性。该 MPEA 是一种基于 MLST 数据中 SNP 的沙门氏菌血清型鉴定和检测的敏感和特异性方法。该方法可以在不到 6 小时的时间内完成，并有可能改善患者护理和溯源。该检测方法用于直接从临床标本和食物样本中鉴定沙门氏菌血清型的效用值得进一步研究。
