Specific detection of Clostridium botulinum types A, B, E, and F using the polymerase chain reaction.
Clostridium botulinum organisms generally produce 1 of 4 neurotoxin types (A, B, E, and F) associated with human illness. Neurotoxin type determination is important in identification of the bacterium. A polymerase chain reaction (PCR) method was developed to identify 24 h botulinal cultures as potential types A, B, E, and F neurotoxin producers as well as other clostridial species which also produce neurotoxins. Components of the PCR and amplification conditions were adjusted for optimal amplification of toxin gene target regions to enable simultaneous testing for types A, B, E, and F in separate tubes using a single thermal cycler. Each primer set was specific for its corresponding toxin type. A DNA extraction procedure was also included to remove inhibitory substances that may affect amplification. This procedure is rapid, sensitive, and specific for identification of toxigenic C. botulinum.
Craven
Kathy E
KE
U.S. Food and Drug Administration, Southeast Regional Laboratory, Atlanta, GA 30309, USA. kcraven@ora.fda.gov
Ferreira
Joseph L
JL
Harrison
Mark A
MA
Edmonds
Paul
P
D001905:Botulinum Toxins / Q000032:analysis*; D003014:Clostridium botulinum / Q000737:chemistry*; D003470:Culture Media; D017931:DNA Primers / Q000032:analysis; D004269:DNA, Bacterial / Q000032:analysis / Q000302:isolation & purification; D007202:Indicators and Reagents; D020133:Reverse Transcriptase Polymerase Chain Reaction
D003470:Culture Media; D017931:DNA Primers; D004269:DNA, Bacterial; D007202:Indicators and Reagents; D001905:Botulinum Toxins