Clostridium perfringens  isolate typing by multiplex PCR

Ahsani MR (1), Mohammadabadi MR (1), Shamsaddini MB (2), Shahabi A (3)

(1) Department of Animal Science, Shahid Bahonar University of Kerman, Kerman, Iran; (2) Razi Vaccine and Serum Research Institute, Kerman, Iran;  (3) Department of Animal Science, Islamic Azad University of Karaj, Tehran, Iran.

ABSTRACT: Clostridium perfringens is an important pathogen that provokes numerous different diseases. This bacterium is classified into five different types, each of which capable of causing a different disease. There are various methods for the bacterium identification, many are labor-intensive, time-consuming, expensive and also present low sensitivity and specificity. The aim of this research was to identify C. perfringens different types using PCR molecular method. In this study, 130 sheep-dung samples were randomly collected from areas around the city of Kerman, southeastern Iran. After processing and culturing of samples, the produced colonies were morphologically studied, gram stain test was also carried out and the genera of these bacteria were identified through biochemical tests. DNA extracted from isolated bacteria for genotyping was tested by multiplex PCR with specific primers. Based on length of synthesized fragments by PCR, toxin types and bacterium strains were detected. C. perfringens isolated types were divided as follows: 17.39% type A, 21.74% type B, 34.78% type C and 26.09% type D. It should be emphasized that, up to the present moment, C. perfringens type A has not been reported in Iran.

KEY WORDS: Clostridium perfringens, multiplex PCR, biochemical tests, genetic typing.

CORRESPONDENCE TO: 

MOHAMMAD REZA AHSANI, Department of Animal Sciences, Shahid Bahonar University of Kerman, 22 Bahman BLVD, Kerman, Kerman, Iran, PO Box: 76169133. Email: Ahsani2001@gmail.com