Comparison of amplified ELISA and mouse bioassay procedures for determination of botulinal toxins A, B, E, and F.
The amplified enzyme-linked immunosorbent assay (amp-ELISA) was compared to the mouse bioassay for determination of botulinal neurotoxin types A, B, E, and F. Twelve different toxin-producing type A, 13 proteolytic type B, 9 nonproteolytic type B, 16 type E, 8 proteolytic type F, 5 nonproteolytic type F, and 6 nontoxigenic clostridial strains were tested. The cultures were inoculated into cooked meat medium (CMM) and tryptone-peptone-glucose-yeast extract (TPGY) medium, incubated for 5 days, and then examined for biological toxicity in mice and amp-ELISA endpoints. The amp-ELISA was less sensitive in detecting toxins produced by nonproteolytic than proteolytic strains of type B and F organisms. All of the toxin-producing strains tested were positive by the AOAC method and the amp-ELISA in either undiluted TPGY or CMM culture fluids regardless of mouse toxicity level, source, or strain. Cross-reactivity was observed between some but not all of the botulinal strains tested. None of the nontoxigenic strains were positive by the amp-ELISA. Purified botulinal toxins were also assayed using these 2 methods. The sensitivity of the amp-ELISA using purified neurotoxins was about 0.1 ng/mL for types A, B, and E and about 1.0 ng/mL for type F.
US Food and Drug Administration, Southeast Regional Laboratory, Atlanta, GA 30309, USA.
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D000818:Animals; D001681:Biological Assay; D001905:Botulinum Toxins / Q000032:analysis*; D019274:Botulinum Toxins, Type A / Q000032:analysis; D003429:Cross Reactions; D003470:Culture Media / Q000032:analysis; D004797:Enzyme-Linked Immunosorbent Assay; D051379:Mice; D012015:Reference Standards
D003470:Culture Media; C096323:rimabotulinumtoxinB; C096325:botulinum toxin type D; D001905:Botulinum Toxins; D019274:Botulinum Toxins, Type A; C096321:botulinum toxin type F