title: Evaluation of a monoclonal antibody-based immunoassay for detecting type A Clostridium botulinum toxin produced in pure culture and an inoculated model cured meat system.
abstract: A monoclonal antibody-based amplified enzyme-linked immunosorbent assay (ELISA) method for detecting Clostridium botulinum type A toxin was evaluated for its ability to detect the toxin in the supernatant fluid of pure cultures and after growth from Cl. botulinum spores inoculated into pork slurries. Slurries containing NaCl (1.5-4.5% w/v) and polyphosphate (0.3% w/v) were either unheated or heated, 80 degrees C/5 min + 70 degrees C/2 h, before storage at 15 degrees, 20 degrees or 27 degrees C. The presence of specific toxin was confirmed by mouse bioassay and results compared with those of the amplified ELISA method. A total of 49 strains, 39 Cl. botulinum and 10 Cl. sporogenes (putrefactive anaerobes), and 95 slurry samples were tested. Fourteen of 15 strains of type A Cl. botulinum and 34 of 36 slurry samples containing type A toxin were positive by ELISA. No false positive reactions occurred with Cl. botulinum types B, C, D, E and F, or with the 10 strains of Cl. sporogenes. However, toxin produced by one strain of Cl. botulinum type A (NCTC 2012) was not detected by the amplified ELISA.
authors.affiliation: Agricultural and Food Research Council, Bristol Laboratory, Langford, UK.
authors.forename: A M
authors.initials: AM
authors.lastname: Gibson
authors.forename: N K
authors.initials: NK
authors.lastname: Modi
authors.forename: T A
authors.initials: TA
authors.lastname: Roberts
authors.forename: C C
authors.initials: CC
authors.lastname: Shone
authors.forename: P
authors.initials: P
authors.lastname: Hambleton
authors.forename: J
authors.initials: J
authors.lastname: Melling
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal; D001905:Botulinum Toxins / Q000032:analysis* / Q000096:biosynthesis; D003014:Clostridium botulinum / Q000302:isolation & purification / Q000378:metabolism*; D004797:Enzyme-Linked Immunosorbent Assay; D005516:Food Microbiology; D008460:Meat; D051379:Mice; D008954:Models, Biological; D013552:Swine; D013696:Temperature
chemical_list: D000911:Antibodies, Monoclonal; D001905:Botulinum Toxins