title: Identification and classification of bcl genes and proteins of Bacillus cereus group organisms and their application in Bacillus anthracis detection and fingerprinting.
abstract: The Bacillus cereus group includes three closely related species, B. anthracis, B. cereus, and B. thuringiensis, which form a highly homogeneous subdivision of the genus Bacillus. One of these species, B. anthracis, has been identified as one of the most probable bacterial biowarfare agents. Here, we evaluate the sequence and length polymorphisms of the Bacillus collagen-like protein bcl genes as a basis for B. anthracis detection and fingerprinting. Five genes, designated bclA to bclE, are present in B. anthracis strains. Examination of bclABCDE sequences identified polymorphisms in bclB alleles of the B. cereus group organisms. These sequence polymorphisms allowed specific detection of B. anthracis strains by PCR using both genomic DNA and purified Bacillus spores in reactions. By exploiting the length variation of the bcl alleles it was demonstrated that the combined bclABCDE PCR products generate markedly different fingerprints for the B. anthracis Ames and Sterne strains. Moreover, we predict that bclABCDE length polymorphism creates unique signatures for B. anthracis strains, which facilitates identification of strains with specificity and confidence. Thus, we present a new diagnostic concept for B. anthracis detection and fingerprinting, which can be used alone or in combination with previously established typing platforms.
authors.affiliation: Nova Research, Inc., Alexandria, Virginia 22308, USA.
authors.forename: Tomasz A
authors.initials: TA
authors.lastname: Leski
authors.forename: Clayton C
authors.initials: CC
authors.lastname: Caswell
authors.forename: Marcin
authors.initials: M
authors.lastname: Pawlowski
authors.forename: David J
authors.initials: DJ
authors.lastname: Klinke
authors.forename: Janusz M
authors.initials: JM
authors.lastname: Bujnicki
authors.forename: Sean J
authors.initials: SJ
authors.lastname: Hart
authors.forename: Slawomir
authors.initials: S
authors.lastname: Lukomski
mesh_terms: D001408:Bacillus anthracis / Q000235:genetics* / Q000302:isolation & purification*; D001409:Bacillus cereus / Q000235:genetics*; D001426:Bacterial Proteins / Q000737:chemistry / Q000235:genetics; D015373:Bacterial Typing Techniques; D016172:DNA Fingerprinting; D014644:Genetic Variation; D008562:Membrane Glycoproteins / Q000737:chemistry / Q000235:genetics*; D008957:Models, Genetic; D011110:Polymorphism, Genetic / Q000235:genetics; D012336:RNA, Ribosomal, 16S / Q000145:classification / Q000235:genetics
chemical_list: D001426:Bacterial Proteins; C470593:BclA protein, Bacillus anthracis; C533739:BclB protein, Bacillus anthracis; D008562:Membrane Glycoproteins; D012336:RNA, Ribosomal, 16S