Characterisation of Clostridium botulinum groups I and II by randomly amplified polymorphic DNA analysis and repetitive element sequence-based PCR

Random amplified polymorphic DNA analysis (RAPD) and repetitive element seq uence-based PCR (rep-PCR) were evaluated with respect to their applicabilit y to characterise Clostridium botulinum group I and II strains, the species causing human botulism. Fifteen group I and 21 group II strains of various geographical and temporal origins were characterised with four single arbi trary RAPD primers at low stringency amplification conditions and with a de generate REP primer pair at moderately stringent conditions. Ready-To-Go RA PD Analysis Beads(TM) and Ready-To-Go PCR Beads(TM) were used for PCR react ions with RAPD and rep-PCR, respectively. Arbitrary primer OPJ 6 yielded th e most discriminating patterns, and distinguished group II C. botulinum ser otypes at the strain level. Group I strains were mainly discriminated at th e serotype level. The discriminatory power of rep-PCR was found to be infer ior to that of RAPD. The REP1R-Dt and REP2R-Dt primer pair generated group I- and II-specific fragments and arbitrary primer OPJ 13 produced a serotyp e E-specific fragment. The use of pre-dispensed and pre-optimised beads att ributed to highly reproducible results. As compared to more time-consuming typing methods, such as pulsed-field gel electrophoresis (PFGE), both RAPD and rep-PCR were characterised by rapid performance and a typeability of 10 0%. (C) 1999 Elsevier Science B.V. All rights reserved.