E. Hyytia et al., Characterisation of Clostridium botulinum groups I and II by randomly amplified polymorphic DNA analysis and repetitive element sequence-based PCR, INT J F MIC, 48(3), 1999, pp. 179-189
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.