Polymerase Chain Reaction for Detection of Type A Clostridium botulinum in Foods

A DNA fragment of the type A Clostridium botulinum neurotoxin gene was demonstrated in canned food products with the polymerase chain reaction (PCR). The fragment, 1340-bp in size, was amplified from green peas, whole kernel corn, green beans, lima beans, black-eyed peas, and turnip greens previously inoculated with type A C. botulinum . The PCR products were identified by agarose gel electrophoresis and also confirmed by dot blot DNA hybridization with a type A specific gene probe. Some inoculated foods were PCR negative but became PCR positive after subculture and overnight incubation in brain heart infusion broth. No PCR amplification products were obtained from uninoculated foods. The procedure was highly sensitive and detected as few as 100 vegetative cells per ml brain heart infusion broth culture.