Chapter title |
High-Throughput CRISPR Typing of Mycobacterium tuberculosis Complex and Salmonella enterica Serotype Typhimurium.
|
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Chapter number | 6 |
Book title |
CRISPR
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Published in |
Methods in molecular biology, January 2015
|
DOI | 10.1007/978-1-4939-2687-9_6 |
Pubmed ID | |
Book ISBNs |
978-1-4939-2686-2, 978-1-4939-2687-9
|
Authors |
Sola, Christophe, Abadia, Edgar, Le Hello, Simon, Weill, François-Xavier, Christophe Sola, Edgar Abadia, Simon Le Hello, François-Xavier Weill, Hello, Simon, Hello, Simon Le |
Editors |
Magnus Lundgren, Emmanuelle Charpentier, Peter C. Fineran |
Abstract |
Spoligotyping was developed almost 18 years ago and still remains a popular first-lane genotyping technique to identify and subtype Mycobacterium tuberculosis complex (MTC) clinical isolates at a phylogeographic level. For other pathogens, such as Salmonella enterica, recent studies suggest that specifically 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 first 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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