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


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.


CONCLUSIONS
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.
Heikinheimo
A
A
Department of Food and Environmental Hygiene, University of Helsinki, Helsinki, Finland. annamari.heikinheimo@helsinki.fi
Korkeala
H
H
D036002:ADP Ribose Transferases / Q000235:genetics; D000818:Animals; D001427:Bacterial Toxins / Q000235:genetics*; D002135:Calcium-Binding Proteins / Q000235:genetics; D002645:Chickens / Q000382:microbiology*; D003015:Clostridium Infections / Q000382:microbiology / Q000662:veterinary; D003016:Clostridium perfringens / Q000235:genetics* / Q000302:isolation & purification; D004269:DNA, Bacterial / Q000032:analysis / Q000302:isolation & purification; D004768:Enterotoxins / Q000235:genetics; D016133:Polymerase Chain Reaction / Q000379:methods; D011201:Poultry Diseases / Q000382:microbiology; D012680:Sensitivity and Specificity; D010738:Type C Phospholipases / Q000235:genetics
D001427:Bacterial Toxins; C031884:CPB protein, Clostridium perfringens; D002135:Calcium-Binding Proteins; C039811:Clostridium perfringens epsilon-toxin; D004269:DNA, Bacterial; D004768:Enterotoxins; C035598:enterotoxin, Clostridium; C050737:iota toxin, Clostridium perfringens; D036002:ADP Ribose Transferases; D010738:Type C Phospholipases; C040649:alpha toxin, Clostridium perfringens