Molecular typing of toxigenic Clostridum perfringens isolated from sheep in Iran

In this research a molecular method based on polymerase chain reaction for typing of Clostridium perfringens was developed and toxin genotypes of ۶۴ isolates from sheep and goats in Iran were determined. 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 were assigned to genotypes A (n=۹, ۱۴.۰۷ %), B (n=۲۰, ۳۱.۲۵%), C (n=۱۷, ۲۶.۵۶%) and D (n=۱۸, ۲۸.۱۲%). In this PCR system the fragments of ۹۰۰, ۶۱۱ and ۴۰۲ 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 ۹۸% 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