Molecular typing of toxigenic Clostridum perfringens isolated from sheep in Iran

Abstract

In this research a molecular method based on polymerase chain reaction for typing of Clostridiumperfringens was developed and toxin genotypes of 64 isolates from sheep and goats in Iran weredetermined. The PCR assays were developed for detection of alpha (cpa), beta (cpb) and epsilon (etx)toxin genes, allowing classification of the isolates into genotypes A B, C and D. The field isolates wereassigned to genotypes A (n=9, 14.07 %), B (n=20, 31.25%), C (n=17, 26.56%) and D (n=18, 28.12%). Inthis PCR system the fragments of 900, 611 and 402 bp were amplified using specific primers for alpha, beta and epsilon toxins, respectively. The fragments were confirmed by sequencing and blasting in GenBank. The sequence alignment of the fragments showed more than 98% similarity with other related published sequences from other sources. Our results suggest that PCR genotyping is an acceptable tool for in vitro typing of C. perfringens.

Keywords

• Clostridium perfringens
• typing
• toxin
• PCR