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全基因组测序为葡萄球菌食物中毒暴发的调查提供了附加价值。

Whole Genome Sequencing Provides an Added Value to the Investigation of Staphylococcal Food Poisoning Outbreaks.

作者信息

Nouws Stéphanie, Bogaerts Bert, Verhaegen Bavo, Denayer Sarah, Laeremans Lasse, Marchal Kathleen, Roosens Nancy H C, Vanneste Kevin, De Keersmaecker Sigrid C J

机构信息

Transversal Activities in Applied Genomics, Sciensano, Brussels, Belgium.

IDLab, Department of Information Technology, Ghent University - IMEC, Ghent, Belgium.

出版信息

Front Microbiol. 2021 Nov 2;12:750278. doi: 10.3389/fmicb.2021.750278. eCollection 2021.

Abstract

Through staphylococcal enterotoxin (SE) production, is a common cause of food poisoning. Detection of staphylococcal food poisoning (SFP) is mostly performed using immunoassays, which, however, only detect five of 27 SEs described to date. Polymerase chain reactions are, therefore, frequently used in complement to identify a bigger arsenal of SE at the gene level () but are labor-intensive. Complete profiling of isolates from different sources, i.e., food and human cases, is, however, important to provide an indication of their potential link within foodborne outbreak investigation. In addition to complete gene profiling, relatedness between isolates is determined with more certainty using pulsed-field gel electrophoresis, protein A gene typing and other methods, but these are shown to lack resolution. We evaluated how whole genome sequencing (WGS) can offer a solution to these shortcomings. By WGS analysis of a selection of isolates, including some belonging to a confirmed foodborne outbreak, its added value as the ultimate multiplexing method was demonstrated. In contrast to PCR-based gene detection for which primers are sometimes shown to be non-specific, WGS enabled complete gene profiling with high performance, provided that a database containing reference sequences for all genes was constructed and employed. The custom compiled database and applied parameters were made publicly available in an online user-friendly interface. As an all-in-one approach with high resolution, WGS additionally allowed inferring correct isolate relationships. The different DNA extraction kits that were tested affected neither gene profiling nor relatedness determination, which is interesting for data sharing during SFP outbreak investigation. Although confirming the production of enterotoxins remains important for SFP investigation, we delivered a proof-of-concept that WGS is a valid alternative and/or complementary tool for outbreak investigation.

摘要

通过产生葡萄球菌肠毒素(SE),是食物中毒的常见原因。葡萄球菌食物中毒(SFP)的检测大多使用免疫测定法进行,然而,该方法仅能检测出迄今为止描述的27种SE中的5种。因此,聚合酶链反应经常作为补充手段用于在基因水平上鉴定更多种类的SE(),但该方法 labor-intensive。然而,对来自不同来源(即食品和人类病例)的分离株进行完整的 分析,对于在食源性疾病暴发调查中表明它们之间可能存在的联系至关重要。除了完整的 基因分析外,使用脉冲场凝胶电泳、蛋白A基因分型和其他方法可以更确定地确定分离株之间的相关性,但这些方法显示出分辨率不足。我们评估了全基因组测序(WGS)如何能够解决这些缺点。通过对一系列分离株进行WGS分析,包括一些属于已确认食源性疾病暴发的分离株,证明了其作为终极多重分析方法的附加价值。与基于PCR的 基因检测不同,有时会显示引物具有非特异性,WGS能够以高性能实现完整的 基因分析,前提是构建并使用包含所有 基因参考序列的数据库。定制编译的数据库和应用参数通过在线用户友好界面公开提供。作为一种具有高分辨率的一体化方法,WGS还允许推断正确的分离株关系。测试的不同DNA提取试剂盒对 基因分析和相关性测定均无影响,这对于SFP暴发调查期间的数据共享很有意义。尽管确认肠毒素的产生对于SFP调查仍然很重要,但我们提供了一个概念验证证明,即WGS是暴发调查中一种有效的替代和/或补充工具。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0671/8593433/b07ced25cde0/fmicb-12-750278-g001.jpg

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