An official website of the United States government.
The objectives of this project were to determine if the DuPont Riboprint system can effectively differentiate between types and individual stains of C. botulinum and therefore serve as a tool to help reduce the incidence of foodborne illness.
The Qualicon RiboprinterJ Microbial Characterization System, currently used to provide genetic 'fingerprints' or Riboprint patterns of microbiological isolates, was evaluated for its ability to differentiate between major types and individual strains of Clostridium botulinum. Pure spores of C. botulinum type A, proteolytic type B, nonproteolytic type B, type E were inoculated onto modified anaerobic egg yolk agar and incubated 24 hrs at 35oC. Plates were rinsed with buffer (2mM Tris + 20mM EDTA) to remove cells which were heated for 10 minutes at 80oC, treated with lysing agent, then ribotyped using the Qualic-on Riboprint-erJ utilizing the enzyme EcoRI. Riboprint patterns obtained for 30 strains of the 4 major types of C. botulinum most commonly involved in human foodborne botulism showed good differentiation within the genus.Proteolytic strains yielded the best and most consistent results. Pattern comparisons determined that 16 separate RiboprintJ groups exist amongst the 30 strains and that distinct and multiple banding patterns appear to exist within each major C. botulinum type. This degree of differentiation between strains of C. botulinum will allow this technique to be used in improving the safety of our food supply, in hazard analysis and identification, HACCP monitoring and validation, environmental monitoring, outbreak epidemiology and in inoculation studies. Establishment of a library of C. botulinum data, or genetic banding patterns specific for each strain of C. botulinum will provide crucial epidemiological information in the case of an outbreak of botulism by identifying specific strains and possible identification of the causative agent.