Typing of Toxigenic Isolates of Clostridium perfringens by Multiplex PCR in Ostrich
Subject Areas : CamelE. Zandi 1 , M.R. Mohammadabadi 2 , M. Ezzatkhah 3 , A.K. Esmailizadeh 4
1 - Department of Animal Science, Shahid Bahonar University of Kerman, Kerman, Iran
2 - Department of Animal Science, Shahid Bahonar University of Kerman, Kerman, Iran
3 - Razi Vaccine and Serum Research Institute, Kerman, Iran
4 - Department of Animal Science, Shahid Bahonar University of Kerman, Kerman, Iran
Keywords: Multiplex PCR, biochemical tests, genetic typing, <i>Clostridium perfringens</i>,
Abstract :
Clostridium perfringens is an important pathogen that provokes numerous different diseases. This bacterium is classified into five various types, each of which capable of causing a distinct disease. There are various methods for the bacterial identification, many are labor-intensive, time-consuming, expensive and also show low sensitivity and specificity. The aim of this research was to identify the unlike types of Clostridium perfringens using PCR method. In this study, 120 ostrich-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 bacterial strains were detected. Clostridium perfringens isolated types were divided as follows: 100% type A, 0% type B, 0% type C and 0% type D. It should be emphasized that, up to now, Clostridium perfringens type A has not been reported in Iran.
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