Open AccessRapid and sensitive identification of pathogenic and apathogenic Bacillus anthracis by real‐time PCR
Author(s) -
Ellerbrok Heinz,
Nattermann Herbert,
Özel Muhsin,
Beutin Lothar,
Appel Bernd,
Pauli Georg
Publication year - 2002
Publication title -
fems microbiology letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.899
H-Index - 151
eISSN - 1574-6968
pISSN - 0378-1097
DOI - 10.1111/j.1574-6968.2002.tb11324.x
Subject(s) - bacillus anthracis , taqman , rpob , microbiology and biotechnology , biology , plasmid , real time polymerase chain reaction , spore , enumeration , polymerase chain reaction , anthrax toxin , bacteria , dna , gene , genetics , 16s ribosomal rna , mathematics , combinatorics , fusion protein , recombinant dna
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.