Identification and characterization of Bacillus anthracis by multiplex PCR analysis of sequences on plasmids pXO1 and pXO2 and chromosomal DNA

FEMS Microbiol Lett. 1996 Nov 15;145(1):9-16. doi: 10.1111/j.1574-6968.1996.tb08548.x.


Bacillus anthracis can be identified on the basis of the detection of virulence factor genes located on two plasmids, pXO1 and pXO2. Thus isolates lacking both pXO1 and pXO2 are indistinguishable from closely related B. cereus group bacteria. We developed a multiplex PCR assay for characterization of B. anthracis isolates, and simultaneous confirmation of the species identity independent of plasmid content. The assay amplifies lef, cya, pag (pXO1) and cap (pXO2) genes, and a B. anthracis specific chromosomal marker, giving an easy-to-read profile. This system unambiguously identified virulent (pXO1+/2+) and avirulent (pXO1+/2-, pXO1-/2+ and pXO1-/2-) strains of B. anthracis and distinguished "anthrax-like' strains from other B. cereus group bacteria.

Publication types

  • Comparative Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Bacillus anthracis / genetics*
  • Bacillus anthracis / isolation & purification
  • Bacillus anthracis / pathogenicity
  • Chromosomes / genetics
  • DNA Primers
  • DNA, Bacterial / analysis
  • Plasmids*
  • Polymerase Chain Reaction
  • Sensitivity and Specificity
  • Virulence


  • DNA Primers
  • DNA, Bacterial