Sciensano, Infectious Diseases in Humans, Bacterial Diseases, B-1180 Brussels, Belgium; Department of Information Technology, IDLab, Imec, Ghent University, B-9052 Ghent, Belgium.
Sciensano, Infectious Diseases in Humans, Bacterial Diseases, B-1180 Brussels, Belgium.
Food Microbiol. 2020 May;87:103394. doi: 10.1016/j.fm.2019.103394. Epub 2019 Nov 25.
Salmonella is a major pathogen having a public health and economic impact in both humans and animals. Six serotypes of the Salmonella genus are mentioned in the Belgian and European regulation as to be rapidly excluded from the food chain (EU regulation N°2160/2003, Belgian royal decree 27/04/2017). The reference method for Salmonella serotyping, including slide-agglutination and biochemical tests, is time-consuming, expensive, not always objective, and therefore does not match the fast identification criteria required by the legislation. In this study, a molecular method, using genetic markers detected by Multiplex Oligonucleotide Ligation - PCR and Luminex technology, was developed for the identification of the 6 Salmonella serotypes and their variants subjected to an official control. The resulting method was validated with the analysis of 971 Salmonella isolated from different matrixes (human, animal, food or environment) and 33 non-Salmonella strains. The results were compared with the reference identifications, achieving an accuracy of 99.7%. The cost-effective high-throughput genoserotyping assay is performed in 1 day and generates objective results, thanks to the automatic interpretation of raw data using a barcode system. In conclusion, it is fully adapted to the implementation in first line laboratories and meets the requirements of the regulation.
沙门氏菌是一种主要的病原体，对人类和动物的公共卫生和经济都有影响。在比利时和欧洲法规中，提到了沙门氏菌属的六种血清型需要从食物链中迅速排除（欧盟法规 2160/2003，比利时皇家法令 27/04/2017）。沙门氏菌血清分型的参考方法，包括玻片凝集和生化试验，既耗时又昂贵，并不总是客观的，因此不符合法规要求的快速鉴定标准。在这项研究中，开发了一种分子方法，使用通过多重寡核苷酸连接 - PCR 和 Luminex 技术检测到的遗传标记，用于鉴定受官方控制的 6 种沙门氏菌血清型及其变体。该方法通过对来自不同基质（人类、动物、食品或环境）的 971 株沙门氏菌和 33 株非沙门氏菌菌株的分析进行了验证。结果与参考鉴定进行了比较，准确率达到 99.7%。这种具有成本效益的高通量基因分型检测可以在 1 天内完成，并由于使用条码系统自动解释原始数据，因此可以生成客观的结果。总之，它完全适用于一线实验室的实施，并满足法规的要求。
