Molecular characterization of Bacillus anthracis using multiplex PCR, ERIC-PCR and RAPD.
OBJECTIVE
To investigate the molecular characterization of Bacillus anthracis strains by multiplex PCR, enterobacterial repetitive intergenic consensus-PCR (ERIC-PCR) and random amplification of polymorphic DNA (RAPD).


RESULTS
Three primers were used to amplify the cya, cap and cereolysinAB genes in the multiplex PCR. Two distinct ERIC-PCR and RAPD fragments, which separated B. anthracis into two groups, were used as probes in Southern hybridization experiments. The probes hybridized only to the cya+ B. anthracis strains identified by the multiplex PCR. Nucleotide sequence analysis of the two cloned fragments showed they were from the pXO1 plasmid of B. anthracis.


CONCLUSIONS
Multiplex PCR simultaneously identified isolates of the Bacillus cereus group and the B. anthracis virulence factors. ERIC-PCR and RAPD, combined with the Southern hybridization analyses, differentiated B. anthracis strains and separated them from the closely related B. cereus group bacteria.


CONCLUSIONS
ERIC-PCR and RAPD assay could be effective in differentiating virulent from avirulent B. anthracis. Our results also show that the amplification of the large plasmids was allowed in the ERIC-PCR and RAPD assay.
Shangkuan
Y H
YH
Division of Bacteriology, Institute of Preventive Medicine, National Defense Medical Center, Taipei, Taiwan, ROC. yhshang@tpts6.seed.net.tw
Chang
Y H
YH
Yang
J F
JF
Lin
H C
HC
Shaio
M F
MF
D001408:Bacillus anthracis / Q000145:classification / Q000235:genetics* / Q000472:pathogenicity; D001409:Bacillus cereus / Q000145:classification / Q000235:genetics / Q000472:pathogenicity; D004269:DNA, Bacterial / Q000032:analysis*; D006801:Humans; D016133:Polymerase Chain Reaction / Q000379:methods; D019105:Random Amplified Polymorphic DNA Technique; D014774:Virulence
D004269:DNA, Bacterial