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. 1966 Mar;91(3):967–974. doi: 10.1128/jb.91.3.967-974.1966

Serological Studies of Types A, B, and E Botulinal Toxins by Passive Hemagglutination and Bentonite Flocculation

H M Johnson 1, K Brenner 1, R Angelotti 1, H E Hall 1
PMCID: PMC315986  PMID: 5326104

Abstract

Johnson, H. M. (Robert A. Taft Sanitary Engineering Center, Cincinnati, Ohio), K. Brenner, R. Angelotti, and H. E. Hall. Serological studies of types A, B, and E botulinal toxins by passive hemagglutination and bentonite flocculation. J. Bacteriol. 91:967–974. 1966.—Formalinized sheep red blood cells (SRBC), sensitized with types A, B, and E botulinal toxoids and toxins by bis-diazotized benzidine (BDB), were tested against A, B, and E antitoxins prepared in horses and rabbits. Type B antitoxin cross-reacted with A toxoid SRBC, but the reciprocal cross-reaction was not observed. E toxin SRBC were specifically agglutinated by E antitoxin. Flocculation of antigen-sensitized bentonite particles was less sensitive in titration of antitoxin than hemagglutination. Also, reciprocal cross-reactions were observed between types A and B antitoxins. Cross-reactions in both serological tests were eliminated by titration of antitoxins in the presence of the heterologous antigens, with no inhibitory effect on the homologous antitoxins. Generally, equine antitoxins were less suitable for agglutinations, especially of antigen-sensitized bentonite particles. Types A, B, and E antitoxins were specifically inhibited by 43, 39, and 245 mouse ld50 of their respective homologous toxins in the hemagglutination-inhibition test. A, B, and E antitoxins were specifically inhibited by 500, 950, and 1,500 mouse ld50 of their respective homologous toxins in bentonite flocculation inhibitions. Formalinized SRBC sensitized with rabbit types A and B antitoxins by BDB were respectively clumped by as little as 0.75 to 1.3 mouse ld50 of A toxin and 2.3 ld50 of B toxin, whereas bentonite particles sensitized by the same antitoxins were specifically clumped by 150 ld50 of A toxin and 630 ld50 of B toxin. E antitoxin sensitization of SRBC or bentonite particles was not successful. Evidence is presented that indicates that the serological procedures are applicable to the detection of botulinal toxins in food.

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Selected References

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  1. BOROFF D. A. Study of toxins of Clostridium botulinum. III. Relation of autolysis to toxin production. J Bacteriol. 1955 Oct;70(4):363–367. doi: 10.1128/jb.70.4.363-367.1955. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. BOWMER E. J. PREPARATION AND ASSAY OF THE INTERNATIONAL STANDARDS FOR CLOSTRIDIUM BOTULINUM TYPES A, B, C, D AND E ANTITOXINS. Bull World Health Organ. 1963;29:701–709. [PMC free article] [PubMed] [Google Scholar]
  3. BOZICEVICH J., NASOU J. P., KAYHOE D. E. Desoxyribonucleic acid (DNA)-bentonite flocculation test for lupus erythematosus. Proc Soc Exp Biol Med. 1960 Mar;103:636–640. doi: 10.3181/00379727-103-25620. [DOI] [PubMed] [Google Scholar]
  4. BUTLER W. T. HEMAGGLUTINATION STUDIES WITH FORMALINIZED ERYTHROCYTES. EFFECT OF BIS-DIAZO-BENZIDINE AND TANNIC ACID TREATMENT ON SENSITIZATION BY SOLUBLE ANTIGEN. J Immunol. 1963 May;90:663–671. [PubMed] [Google Scholar]
  5. CRISLEY F. D. Routine method for the goldfish assay of toxin in crude culture centrifugates of Clostridium botulinum type A. Appl Microbiol. 1960 Sep;8:282–285. doi: 10.1128/am.8.5.282-285.1960. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. DUFF J. T., WRIGHT G. G., KLERER J., MOORE D. E., BIBLER R. H. Studies on immunity to toxins of Clostridium botulinum. I. A simplified procedure for isolation of type A toxin. J Bacteriol. 1957 Jan;73(1):42–47. doi: 10.1128/jb.73.1.42-47.1957. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. GORDON J., ROSE B., SEHON A. H. Detection of non-precipitating antibodies in sera of individuals allergic to ragweed pollen by an in vitro method. J Exp Med. 1958 Jul 1;108(1):37–51. doi: 10.1084/jem.108.1.37. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. LAMANNA C., LOWENTHAL J. P. The lack of identity between hemagglutinin and the toxin of type A botulinal organism. J Bacteriol. 1951 Jun;61(6):751–752. doi: 10.1128/jb.61.6.751-752.1951. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. PETTY C. S. BOTULISM: THE DISEASE AND THE TOXIN. Am J Med Sci. 1965 Mar;249:345–359. [PubMed] [Google Scholar]
  10. SINITSYN V. A. [The use of the indirect hemagglutination reaction for detection of botulinus toxin]. Voen Med Zh. 1961 Oct;10:65–67. [PubMed] [Google Scholar]
  11. STAVITSKY A. B. Micromethods for the study of proteins and antibodies. I. Procedure and general applications of hemagglutination and hemagglutination-inhibition reactions with tannic acid and protein-treated red blood cells. J Immunol. 1954 May;72(5):360–367. [PubMed] [Google Scholar]
  12. WOLFF S. M., WARD S. B., LANDY M. SEROLOGIC PROPERTIES OF BENTONITE PARTICLES COATED WITH MICROBIAL POLYSACCHARIDES. Proc Soc Exp Biol Med. 1963 Nov;114:530–536. doi: 10.3181/00379727-114-28724. [DOI] [PubMed] [Google Scholar]

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