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Lett Appl Microbiol. 2001 Mar;32(3):139-45.

Molecular characterization of Bacillus anthracis using multiplex PCR, ERIC-PCR and RAPD.

Author information

1
Division of Bacteriology, Institute of Preventive Medicine, National Defense Medical Center, Taipei, Taiwan, ROC. yhshang@tpts6.seed.net.tw

Abstract

AIMS:

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).

METHODS AND 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.

CONCLUSION:

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.

SIGNIFICANCE AND IMPACT OF THE STUDY:

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.

PMID:
11264741
[Indexed for MEDLINE]
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