Comparative analysis of nontoxigenic and neurotoxigenic Clostridium butyricum by molecular typing methods (original) (raw)

2000

Abstract

ABSTRACT Neurotoxigenic Clostridium butyricum has been identified as occurring in the natural environment. Nontoxigenic and neurotoxigenic C. butyricum strains were comparatively analyzed by PCR assay and Southern blot hybridization for the type E botulinum toxin gene (bont/E), random amplified polymorphic DNA (RAPD) assay, and pulsed-field gel electrophoresis (PFGE). With the PCR assay and Southern blot hybridization, the bont/E gene was detected in all seven strains of neurotoxigenic C. butyricum (BL 5262, BL 6340, LCL 155, LCL 063, LCL 095, KZ 1886, and KZ 1887), but not in nontoxigenic strains (MIYAIRI 588, MIYAIRI 595, MIYAIRI 630, SI 293-2, RU 063-3, GU-2, ATCC 19398, and IFO 3315), indicating that there were no partial bont/E gene fragments in the nontoxigenic C. butyricum strains. All strains were successfully analyzed by RAPD assay. In contrast to the RAPD assay, two strains of nontoxigenic C. butyricum could not be analyzed by PFGE, probably due to the DNase activity. Nontoxigenic strains SI 293-2 and GU-2 shared an identical RAPD or PFGE pattern. The other nontoxigenic strains showed unique RAPD and PFGE patterns, and these patterns differed from those of all neurotoxigenic C. butyricum strains. The present results are discussed in relation to the transfer of the bont/E gene.

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