Comparison of Toxicity Neutralization-, ELISA- and PCR Tests for Typing of Clostridium perfringensand Detection of the Enterotoxin Gene by PCR

Abstract

A polymerase chain reaction (PCR) was developed for the specific amplification of a part of each of the five Clostridium perfringenstoxin genes: alpha (α), beta (β), epsilon (ϵ), iota (ι), and enterotoxin (CPE). While the toxicity neutralization test (TNT) only showed limited ability to detect the α toxin, the lecithinase test and PCR test (PCR α) concordantly detected the α toxin and the α toxin gene, respectively. A monoclonal enzyme linked immunosorbent assay (ELISA) and a PCR βtest were compared and were in accordance for the detection of the β toxin (gene) from pure and mixed cultures from piglets suffering from necrotizing enteritis. However, the PCR βtest was superior to the ELISA for detection of the β toxin (gene) in necrotic intestinal mucosa without culturing. An internal standard to be co-amplified with the β toxin gene was constructed and served as a control for inhibition of the PCR βtest. The enterotoxin gene was not detected in any of 95 Danish Clostridium perfringensfield isolates. This indicates that the C. perfringensenterotoxin is not involved in diarrhoea in certain animal species from this area. The origin of enterotoxin-positive C. perfringensinvolved in intoxication of humans will need special attention in future studies.