Identification and classification of bcl genes and proteins of Bacillus cereus group organisms and their application in Bacillus anthracis detection and fingerprinting.
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.
Leski
Tomasz A
TA
Nova Research, Inc., Alexandria, Virginia 22308, USA.
Caswell
Clayton C
CC
Pawlowski
Marcin
M
Klinke
David J
DJ
Bujnicki
Janusz M
JM
Hart
Sean J
SJ
Lukomski
Slawomir
S
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
D001426:Bacterial Proteins; C470593:BclA protein, Bacillus anthracis; C533739:BclB protein, Bacillus anthracis; D008562:Membrane Glycoproteins; D012336:RNA, Ribosomal, 16S