Detection of Botulinal Toxins by Immunodiffusion

Abstract

A procedure employing concentration with Sephadex and analysis by gel diffusion (Ouchterlony) was used to detect the toxins of Clostridium botulinum in foods. Botulinal toxins with toxic levels of 370 to 557 mouse LD 50 per milliliter were detected in the food samples. Test results were verified by use of the mouse protection test. Approximately 24 hr were required to complete the entire procedure.

Selection and Development of Nontoxic Nonproteolytic Clostridium botulinum Surrogate Strains for Food Challenge Testing

“…Botulinum selective medium (BSM), a medium containing egg yolk, cycloserine, sulfamethoxazole, and trimethoprim, was developed decades ago but does not suppress or allow to distinguish several other clostridial species and, importantly, inhibits some nonproteolytic C. botulinum strains [ 31 , 32 ]. The inclusion of antitoxin antibodies in the growth medium was shown to improve the detection of BoNT-producing strains by the formation of a precipitation zone around the colonies, but no data is available on the performance of these plates with complex samples containing background microbiota [ 33 , 34 ]. Finally, fluorescent antibodies against vegetative cell walls of C. botulinum have been used in culture, but this technique has never been adopted for use in solid media to determine C. botulinum cell counts [ 35 ].…”

Untersuchungen zu Auftreten von Clostridium botulinum, betriebsspezifischen Risikofaktoren und Symptomen beim Krankheitsbild des viszeralen Botulismus

“…Durch das gleichzeitige Herauslösen von Stoffen, die inhibierend wirken, kann die Amplifikation wesentlich beeinträchtigt werden (Bessetti, 2007) (Higuchi et al, 1992;Heid et al, 1996). Farbstoffe wie Ethidiumbromid oder SYBR ® Green interkalieren in die doppelsträngige DNA, wodurch die Fluoreszenz dieser (Campbell et al, 1993b) x x x (Fach et al, 1995) x x x x x (Kimura et al, 2001) x (Akbulut et al, 2004) x x x (Yoon et al, 2005) x (Artin et al, 2007) x (Fenicia et al, 2007) x (Raphael und Andreadis, 2007) x (Fach et al, 2009 (Kozaki et al, 1974;Gill, 1982) (Simpson, 1971;Simpson, 1973;Simpson, 1974 (Boroff und Shu-Chen, 1973;Ashton et al, 1985), der Geldiffusionsassay (Vermilyea et al, 1968;Miller und Anderson, 1971;Ferreira et al, 1981) oder auch der passive Hämagglutinationsassay (Johnson et al, 1966) In der Botulismus-Diagnostik sind ELISAs für reines BoNT (Ekong et al, 1995;Szílagyi et al, 2000;Poli et al, 2002), toxische C.-botulinum-Kulturen (Dezfulian und Bartlett, 1984;Dezfulian und Bartlett, 1985;Shone et al, 1985;Schrödl et al, 2007), Lebensmittel (Doellgast et al, 1993;Rodriguez und Dezfulian, 1997;Ferreira und Crawford, 1998), Serum (Poli et al, 2002) Yoon et al, 2004;Hörman et al, 2005;Sharma et al, 2005;…”