Staphylococcal Enterotoxin D Immunological Identification with Purified Toxin

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Purification of enterotoxin D from Staphylococcus aureus 494 was attempted by utilizing the following techniques: Concentration of bacterial culture supernatant with polyethylene glycol 20000, CM-cellulose column chromatography, Sephadex G-100 gel-filtration, chromatography on a DEAE cellulose column with concave gradient elution and gel-filtration with Sephadex G-50. The final preparation obtained gave a single precipitin line with anti-crude enterotoxin D serum and no precipitation with anti-enterotoxin A, B or C when it was tested by Ouchterlony's technique. It was capable of producing 100% emesis in monkeys at about 1.25μg of protein per kg body weight. Antiserum prepared by injecting rabbits with the purified preparation gave a single precipitin line with concentrated crude toxin, and it also gave a single are when tested in immuncelectrophoresis with concentrated crude toxin. A type-specific neutralization effect in monkeys was obtained. Thus, enterotoxin D has been identified with a purified preparation.