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

Christophe Sola (1) , Edgar Abadia (2) , Simon Le Hello (3) , François-Xavier Weill (3)

1 I2BC - Institut de Biologie Intégrative de la Cellule
2 IVIC - Instituto Venezolano de Investigaciones Cientificas
3 CNR-ESS - Centre National de Référence - National Reference Center Escherichia coli, Shigella et Salmonella

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.

Keywords: CRISPR locus; Spoligotyping; High-throughput; Molecular epidemiology; Microbeads

Domains: Life Sciences [q-bio]; Santé publique et épidémiologie

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, 2015, CRISPR : Methods and Protocols, 1311, pp.91-109. ⟨10.1007/978-1-4939-2687-9_6⟩. ⟨pasteur-01159705⟩