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Typage et sous-typage du genre Salmonella basé sur le polymorphisme des loci CRISPR

Abstract : Salmonella is the first causative agent of bacterial diarrhea due to food poisoning in developed countries and is still considered as a major public health problem. For almost a century, serotyping has been the gold standard method for Salmonella identification. Based on the identification of somatic and flagellar antigens, this manual technique remains time consuming (3-6 days) and expensive (requires the use of > 150 manufactured antisera). About 2 600 serotypes have been described within the Salmonella genus. Their distribution being the basis for epidemiological investigations, a fast and accurate identification is essential. Moreover, due to the predominance of particular serotypes, an additional step of subtyping (Pulse-Field Gel Electrophoresis-PFGE) is necessary for a better discrimination and the identification of epidemic clones (5 more days after serotyping). In order to optimize epidemiological investigations, we need to develop a new high throughput method for fast and accurate typing/subtyping, which led us to study the polymorphism of the CRISPR (Clustered Regularly Interspaced Short Palindromic Repeats) region in Salmonella. Our study on 134 serotypes highlighted the strong correlation between spacer content and serotype/subtype. We also developed a high-throughput method to subtype serotype Typhimurium (CRISPOL), a new PCR method for fast identification of serotypes Typhi and Paratyphi A and a novel PCR method for fast detection of ST198 population of serotype Kentucky.The discriminatory power of CRISPR typing offers a new prospective for salmonellosis monitoring with a fast and accurate method that could easily replace gold standards serotyping and PFGE.
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Submitted on : Tuesday, October 19, 2021 - 12:00:18 PM
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Laetitia Fabre. Typage et sous-typage du genre Salmonella basé sur le polymorphisme des loci CRISPR. Microbiologie et Parasitologie. Institut agronomique, vétérinaire et forestier de France, 2017. Français. ⟨NNT : 2017IAVF0002⟩. ⟨tel-03385008⟩

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