Multiplex PCR assay for toxinotyping Clostridium perfringens isolates obtained from Finnish broiler chickens


Annamari Heikinheimo, PO Box 66, FIN-00014, Helsinki University, Finland (e-mail:


Aims:  The aim of the study was to determine the presence of genes coding for alpha (cpa), beta (cpb), epsilon (etx), iota (iA) and enterotoxin (cpe) from Clostridium perfringens broiler chicken isolates, using multiplex PCR assay established in the study.

Methods and Results:  The multiplex PCR assay was shown to be specific when tested with 10 C. perfringens strains representing different toxin types, and 15 strains of other bacterial species. All 118 broiler chicken C. perfringens isolates were shown to carry the cpa gene but not cpb, etx, iap or cpe genes, signifying that all isolates represented type A and were cpe-negative.

Conclusions:  The assay established in the study enables the simultaneous detection of the major toxin genes and the cpe gene from C. perfringens isolates.

Significance and Impact of the Study:  The present study offers a new primer pair for detecting cpa, combined with a multiplex PCR assay. In addition, the study provides data of the presence of different toxin genes in C. perfringens isolates obtained from broiler chickens.