Real-time PCR system targeting a chromosomal marker specific for Bacillus anthracis

Markus H. Antwerpen*, Pia Zimmermann, Kevin Bewley, Dimitrios Frangoulidis, Hermann Meyer

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

55 Citations (Scopus)

Abstract

Specific identification of Bacillus anthracis and differentiation from closely related Bacillus cereus and Bacillus thuringiensis strains is still a major diagnostic problem. Commercially available diagnostic kits targeting plasmid-markers cannot differentiate between B. anthracis, non-anthracis Bacillus species harbouring anthrax-specific virulence plasmids, and plasmidless B. anthracis strains. A TaqMan® PCR assay was designed targeting sequences of gene locus BA_5345 of the B. anthracis strain Ames. Specificity was determined by using a panel of 328 Bacillus strains; sensitivity was determined by probit analysis. All B. anthracis isolates (n = 92) were specifically detected by using the genomic TaqMan® PCR assay whereas 236 strains belonging to 19 Bacillus species other than B. anthracis were PCR negative. The detection limit was determined to be 12.7 copies per reaction (95% confidence interval 10.2-17.5 copies). Here we present an extensively evaluated and - to our current knowledge - specific TaqMan® PCR assay for the detection of B. anthracis based on a chromosomal marker.

Original languageEnglish
Pages (from-to)313-315
Number of pages3
JournalMolecular and Cellular Probes
Volume22
Issue number5-6
DOIs
Publication statusPublished - Oct 2008

Keywords

  • BA5345
  • Bacillus anthracis
  • Chromosomal marker
  • Real-time PCR
  • dhp61

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