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

Methods Mol Biol. 2015:1311:91-109. doi: 10.1007/978-1-4939-2687-9_6.

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.

MeSH terms

  • Clustered Regularly Interspaced Short Palindromic Repeats*
  • Microspheres
  • Molecular Typing / methods*
  • Mycobacterium tuberculosis / classification*
  • Mycobacterium tuberculosis / genetics*
  • Oligonucleotides / chemistry
  • Oligonucleotides / genetics
  • Salmonella typhimurium / classification*
  • Salmonella typhimurium / genetics*

Substances

  • Oligonucleotides