Rapid and sensitive identification of pathogenic and apathogenic Bacillus anthracis by real-time PCR.
Bacillus anthracis spores have been shown to be an efficient biological weapon and their recent use in bioterrorist attacks has demonstrated the need for rapid and specific diagnostics. A TaqMan real-time PCR for identification of B. anthracis was developed, based on the two plasmids, pX01 and pX02, both of which are necessary for pathogenicity, as well as on the chromosomally encoded rpoB gene. Bacteria picked from colonies or pelleted from liquid cultures were directly inoculated into the PCR mix, thus avoiding time-consuming DNA preparation and minimizing handling risks. B. anthracis spores were cultivated for a few hours in enrichment broth before PCR analysis, or used directly for real-time PCR, thus allowing to confirm or exclude potential attacks approximately 2-3 h after the material has arrived in the laboratory.
Robert Koch-Institut, Nordufer 20, 13353 Berlin, Germany. ellerbrokh@rki.de
Heinz
H
Ellerbrok
Herbert
H
Nattermann
Muhsin
M
Ozel
Lothar
L
Beutin
Bernd
B
Appel
Georg
G
Pauli
D000881:Anthrax / Q000175:diagnosis* / Q000382:microbiology; D000942:Antigens, Bacterial; D001408:Bacillus anthracis / Q000145:classification* / Q000235:genetics / Q000472:pathogenicity*; D016667:Bacterial Capsules / Q000235:genetics; D001427:Bacterial Toxins / Q000235:genetics; D015373:Bacterial Typing Techniques; D001694:Biological Warfare; D004269:DNA, Bacterial / Q000032:analysis; D012321:DNA-Directed RNA Polymerases / Q000235:genetics; D006801:Humans; D010957:Plasmids / Q000235:genetics; D016133:Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity; D013045:Species Specificity; D013171:Spores, Bacterial; D019914:Taq Polymerase; D013997:Time Factors; D014774:Virulence / Q000235:genetics
D000942:Antigens, Bacterial; D001427:Bacterial Toxins; D004269:DNA, Bacterial; C030325:anthrax toxin; D019914:Taq Polymerase; D012321:DNA-Directed RNA Polymerases; C105078:RNA polymerase beta subunit