Genome-wide single nucleotide polymorphism typing method for identification of Bacillus anthracis species and strains among B. cereus group species

J Clin Microbiol. 2010 Aug;48(8):2821-9. doi: 10.1128/JCM.00137-10. Epub 2010 Jun 16.

Abstract

As an issue of biosecurity, species-specific genetic markers have been well characterized. However, Bacillus anthracis strain-specific information is currently not sufficient for traceability to identify the origin of the strain. By using genome-wide screening using short read mapping, we identified strain-specific single nucleotide polymorphisms (SNPs) among B. anthracis strains including Japanese isolates, and we further developed a simplified 80-tag SNP typing method for the primary investigation of traceability. These 80-tag SNPs were selected from 2,965 SNPs on the chromosome and the pXO1 and pXO2 plasmids from a total of 19 B. anthracis strains, including the available genome sequences of 17 strains in the GenBank database and 2 Japanese isolates that were sequenced in this study. Phylogenetic analysis based on 80-tag SNP typing showed a higher resolution power to discriminate 12 Japanese isolates rather than the 25 loci identified by multiple-locus variable-number tandem-repeat analysis (MLVA). In addition, the 80-tag PCR testing enabled the discrimination of B. anthracis from other B. cereus group species, helping to identify whether a suspected sample originates from the intentional release of a bioterrorism agent or environmental contamination with a virulent agent. In conclusion, 80-tag SNP typing can be a rapid and sufficient test for the primary investigation of strain origin. Subsequent whole-genome sequencing will reveal apparent strain-specific genetic markers for traceability of strains following an anthrax outbreak.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Bacillus anthracis / classification*
  • Bacillus anthracis / genetics*
  • Bacillus anthracis / isolation & purification
  • Bacillus cereus / classification*
  • Bacillus cereus / genetics*
  • Bacillus cereus / isolation & purification
  • Bacterial Typing Techniques / methods*
  • Chromosomes, Bacterial
  • DNA, Bacterial / genetics
  • Genome, Bacterial
  • Genotype
  • Humans
  • Japan
  • Phylogeny
  • Plasmids
  • Polymerase Chain Reaction / methods*
  • Polymorphism, Single Nucleotide*

Substances

  • DNA, Bacterial