Detection of Clostridium botulinum type A toxin by enzyme-linked immunosorbent assay with antibodies produced in immunologically tolerant animals

Abstract

Immunological tolerance is a state of unresponsiveness to foreign substances (antigens) which can develop in human and animal species as the result of continued exposure to antigens early in life. We utilized this principle for the preparation of antibodies against Clostridium botulinum type A toxin. By selective suppression of the immunological response of rabbits to unwanted antigens and subsequent immunization with a toxoid, we were able to produce a specific type A antitoxin without the need to purify the toxin. Despite cross-reactivity with C. botulinum type B, our type A antitoxin was otherwise specific since it did not react with culture filtrates of nontoxigenic variants of type B, any other C. botulinum type (C, D, E, F, and G), nor with 18 other Clostridium species, including Clostridium sporogenes. Using this antitoxin, we developed a sensitive enzyme-linked immunosorbent assay for detection of C. botulinum type A toxin.

Quantification of Factors Which Influence Nisin's Inhibition of CIostridium botulinum 56A in a Model Food System

“…We considered these results to be false negatives because if toxin was produced in the presence of nisin, it should also have been produced in its absence. We attribute these few false-negatives to the lack of sensitivity in the ELISA method which is about lo-fold less sensitive than the mouse assay (Dezfulian and Bartlett, 1984). We have observed similar results for borderline detectable amounts of toxin in a model gravy system (Crandall and Montville, 1993).…”

“…based on Dezfulian and Bartlett (1984). From the results of the preliminary experiments, protein, phospholipid, and soluble starch were selected for inclusion in the main modeling experiment.…”

Enzyme-linked immunosorbent assay for detection of Clostridium botulinum type A and type B toxins in stool samples of infants with botulism

“…Antigens to produce these antitoxins in burros were the partially purified toxoid prepared for human immunization (3). Rabbit antitoxins specific for botulinal toxins A and B were prepared by the method described previously (5). The optimal dilutions of rabbit and burro antitoxins used for coating wells for the ELISA were determined by checkerboard titration.…”

Die Behandlung von Dystonien mit Botulinumtoxin

“…1 Indikationen zur Behandlung mit BTA.Periphere Wirkung: BTA hemmt die Freisetzung von Azetyicholin an der neuromuskularen Endplatte und induziert je nach Dosierung eine unterschiedlich schwere Muskelschwäche, die bei lokaler Injektion weitgehend auf den behandelten Muskel beschränkt Inwieweit Immunisierung, d. h. Antikorperbildung, weiche sich derzeit nur in biologischen Assays zuverlassig nachweisen IaBt, eine Rolle spielt, 1st noch offen. Generell kann man sagen, dad BTA em schlechtes Antigen ist(9,23,27).NebenwirkungenHauptnebenwirkung 1st eine zu starke Abschwachung der injizierten oder eine unerwunschte Mitbeteiligung benachbarter, nicht behandelter Muskein, die wahrscheinlich durch Diffusion des Toxins bedingt ist. Da die Nebenwirkungen gleichsam die Spitze eines Eisberges darstellen, klingen sie wesentlich rascher ab als die erwunschte Wirkung und sind meist nach 3 his vier Wochen verschwunden.…”