Development of a novel, rapid, in vitro assay for the detection of Clostridium botulinum neurotoxin type E

• Main Author: Cadieux, Brigitte.
• Other Authors: Smith, J. P. (advisor)
• Format: Others
• Language: en
• Published: McGill University 2001
• Subjects: Clostridium botulinum.; Botulinum toxin.; Toxicity testing > In vitro.
• Online Access: http://digitool.Library.McGill.CA:80/R/?func=dbin-jump-full&object_id=32836

Botulism is a foodborne intoxication caused by ingestion of Clostridium botulinum neurotoxin (BoNT). Preliminary studies focussed on the production of polyclonal antisera against BoNT/E by immunizing a rabbit with botulinal toxoid type E. The antiserum was subsequently used to detect BoNT/E using the slot blot immunoassay where samples were applied to a slot blot filtration manifold and drawn by vacuum through a membrane. The membrane was then immunologically processed before chemiluminescent detection. However, the antisera lacked specificity and cross-reacted with closely related clostridia strains. === The specificity of the antisera was increased by adsorbing cross-reactive antibodies from whole antisera with affinity columns made with total proteins from culture supernatants of closely related clostridia. Alternatively, specific antibodies were isolated with an affinity column prepared with C. botulinum type E toxoid. === Different methods of concentrating BoNT/E in each sample prior to testing them were evaluated to increase the sensitivity of the assay. === The slot blot immunoassay was then evaluated for detection of BoNT/E in mixed cultures and in food samples. (Abstract shortened by UMI.)