Assay Methods for Clostridium perfringens Type A Enterotoxin

Abstract

Enterotoxin produced by a sporulating culture of Clostridium perfringens type A NCTC 8798 was purified to a level of 3,500 mouse mean lethal doses per mg of nitrogen. High-titer sera were obtained from rabbits injected with enterotoxin and used to compare the sensitivity of serological tests and bioassays for C. perfringens enterotoxin. Reversed passive hemagglutination was by far the most sensitive test, followed by microslide diffusion, single gel diffusion and electroimmunodiffusion, guinea pig skin test, mouse test, and rabbit ileal loop test.

Measurement of biological activities of purified and crude enterotoxin of Clostridium perfringens

“…Toxicity of the pure enterotoxin for mice when injected i.v. and corrected for body weight was 684 LD,0/mg of protein, which compares with 556 LD,0/mg of protein obtained by Genigeorgis et al (3). Since the latter value was not weight-corrected certain differences can be expected, since the response of the animals to i.v.-injected enterotoxin is dependent on the body weight (10).…”

Response of Ligated Rabbit Ileal Loop to Clostridium perfringens Type C Strains and Their Toxic Filtrates

“…Antitoxin sera of C. perfringens types A, B, C, D, and E were purchased from Wellcome Research Laboratory, Bechenham, England. Antisera against the enterotoxins of C. perfringens types A and C were prepared by the method of Genigeorgis et al (5).…”

The Suitability of Tórtora's Medium for the Production of Enterotoxin in Clostridium perfringens Strains

“…Purification of the enterotoxin was carried out as described by Granum and Whitaker (12) ; however the chromatography on a Sephadex column was omitted. The partially purified enterotoxin was injected subcutaneously into rabbits by the method of Genigeorgis et al (11). The sera obtained showed a titer between 1 :50 and 1:80 in microslide double immunodiffusion tests (21) and showed enterotoxin specificity when diluted 1:10 or more in sterile NaCl (0.85% NOT) solution.…”