Distribution of minor virulence genes among Clostridium perfringens Isolates

Clostridium perfringens (C. perfringens) belongs to the family of Clostridiaceae and produces a wide range of toxins (four major and a variety of minor toxins). Some toxins associated with virulence have been shown to participate in the pathogenesis of enteric diseases in sheep, goats, and other animals. The aim of this study was to determine the presence of some minor virulence genes and their genetic diversity in C. perfringens isolates.About ۸۴ isolates collected from sheep and goat flocks were provided by the microbial archive of Razi Institute (south-east branch) that located in Kerman. Isolates were removed from the cryotube stored at -۷۰° C; then, it smeared on blood agar medium containing ۵ % defibrinated sheep blood, which was incubated in an anaerobic condition.After DNA extraction, detection of toxin genes (cpb۲, tpel, and PFO) was carried out, using three pairs of specific primers were examined and sequenced for the presence of minor virulence genes( PFO , cpb۲ and tpel ) by PCR method. Purified PCR products were sequenced using the Sinacolon facility, Tehran, Iran. The gene sequences were aligned according to their nucleotides, using computer program MEGA ۷. Then, analysis was performed for confirm the nucleotide sequences.Results showed that PFO and cpb۲ were found in ۷۹ out of ۸۴ (۹۴.۴ %) and the presence of tpel was confirmed in ۲۹ out of ۸۴ (۳۵ %) isolates so the dominant minor virulence genes were PFO and cpb۲. prevalence of these genes in C. perfringens isolates would provide more information regarding the importance of these toxins and lead to a greater understanding of the pathogenesis of diseases caused by C. perfringens, furthermore DNA sequencing revealed closed relationships with others world strains that were range approximately (۹۷−۱۰۰ %) with the GenBank database.

کلیدواژه ها:

C. perfringens ، minor virulence genes ، Sequence ، PCR