Identification and characterization of Bacillus anthracis by multiplex PCR analysis of sequences on plasmids pXO1 and pXO2 and chromosomal DNA |
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| Authors: | Vincent Ramisse,Guy Patra,Henri Garrigue,Jean-Luc Guesdon,Michè le Mock |
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| Affiliation: | Laboratoire de Microbiologie Appliquée, Centre d'Etudes du Bouchet, B.P. no. 3, 91710 Vert-Le-Petit, France; Service Biotechnologies, Centre d'Etudes du Bouchet, B.P. no. 3, 91710 Vert-Le-Petit, France; Laboratoire de Prédéveloppement des Sondes, Institut Pasteur, 75724 Paris Cedex 15, France; Laboratoire de Génétique Moléculaire des Toxines, CNRS URA 1858, Institut Pasteur, 75724 Paris Cedex 15, France |
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| Abstract: | Abstract 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. |
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| Keywords: | Bacillaceae Bacillus anthracis Bacillus cereus Bacillus anthracis genetics Anthrax epidemiology Bacterial identification Diagnosis Polymerase chain reaction |
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