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High-Throughput CRISPR Typing of Mycobacterium tuberculosis Complex and Salmonella enterica Serotype Typhimurium

Abstract : Spoligotyping was developed almost 18 years ago and still remains a popular fi rst-lane genotyping technique to identify and subtype Mycobacterium tuberculosis complex (MTC) clinical isolates at a phylogeo-graphic level. For other pathogens, such as Salmonella enterica , recent studies suggest that specifi cally designed spoligotyping techniques could be interesting for public health purposes. Spoligotyping was in its original format a reverse line-blot hybridization method using capture probes designed on " spacers " and attached to a membrane's surface and a PCR product obtained from clustered regularly interspaced short palindromic repeats (CRISPRs). Cowan et al. and Fabre et al. were the fi rst to propose a high-throughput Spoligotyping method based on microbeads for MTC and S. enterica serotype Typhimurium, respectively. The main advantages of the high-throughput Spoligotyping techniques we describe here are their low cost, their robustness, and the existence (at least for MTC) of very large databases that allow comparisons between spoligotypes from anywhere.
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Christophe Sola, Edgar Abadia, Simon Le Hello, François-Xavier Weill. High-Throughput CRISPR Typing of Mycobacterium tuberculosis Complex and Salmonella enterica Serotype Typhimurium. Methods in Molecular Biology, Humana Press/Springer Imprint, 2015, CRISPR : Methods and Protocols, 1311, pp.91-109. ⟨10.1007/978-1-4939-2687-9_6⟩. ⟨pasteur-01159705⟩

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