TY - JOUR
T1 - Real-time PCR system targeting a chromosomal marker specific for Bacillus anthracis
AU - Antwerpen, Markus H.
AU - Zimmermann, Pia
AU - Bewley, Kevin
AU - Frangoulidis, Dimitrios
AU - Meyer, Hermann
PY - 2008/10
Y1 - 2008/10
N2 - 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.
AB - 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.
KW - BA5345
KW - Bacillus anthracis
KW - Chromosomal marker
KW - Real-time PCR
KW - dhp61
UR - http://www.scopus.com/inward/record.url?scp=56749106410&partnerID=8YFLogxK
U2 - 10.1016/j.mcp.2008.06.001
DO - 10.1016/j.mcp.2008.06.001
M3 - Article
C2 - 18602986
AN - SCOPUS:56749106410
SN - 0890-8508
VL - 22
SP - 313
EP - 315
JO - Molecular and Cellular Probes
JF - Molecular and Cellular Probes
IS - 5-6
ER -