COMPARISON OF TOXICITY NEUTRALIZATION-TEST, ELISA-TEST AND PCR-TEST FOR TYPING OF CLOSTRIDIUM-PERFRINGENS AND DETECTION OF THE ENTEROTOXIN GENE BY PCR

A polymerase chain reaction (PCR) was developed for the specific ampli fication of a part of each of the five Clostridium perfringens toxin g enes: alpha (alpha), beta (beta), epsilon (epsilon), iota (iota), and enterotoxin (CPE). While the toxicity neutralization test (TNT) only s howed limited ability to detect the or toxin, the lecithinase test and PCR test (PCR(alpha)) concordantly detected the ct toxin and the alph a toxin gene, respectively. A monoclonal enzyme linked immunosorbent a ssay (ELISA) and a PCR(beta) test were compared and were in accordance for the detection of the beta toxin (gene) from pure and mixed cultur es from piglets suffering from necrotizing enteritis. However, the PCR (beta) test was superior to the ELISA for detection of the beta toxin (gene) in necrotic intestinal mucosa without culturing. An internal st andard to be co-amplified with the beta toxin gene was constructed and served as a control for inhibition of the PCR(beta) test. The enterot oxin gene was not in any of 95 Danish Clostridium perfringens field is olates. This indicates that the C. perfringens enterotoxin is not invo lved in diarrhoea in certain animal species from this area. The origin of enterotoxin-positive C. perfringens involved in intoxication of hu mans will need special attention in future studies. (C) 1996 Academic Press