Clostridium perfringens isolate typing by multiplex PCR

Mr Ahsani,Mb Shamsaddini,Mr Mohammadabadi

doi:10.1590/s1678-91992010000400006

Mr Ahsani, Mb Shamsaddini + Show 1 more

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https://doi.org/10.1590/s1678-91992010000400006

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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 bacterial identification, many are labor-intensive, time-consuming, expensive and also present low sensitivity and specificity. The aim of this research was to identify the different types of C. perfringens 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 bacterial 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.

Highlights

C. perfringens has a notorious reputation for possessing an extremely wide toxin arsenal, with over 15 toxins and may be the most widely occurring pathogenic in the globe [1, 2]
The DNA extracted from all bacterial strains was identified by biochemical tests and standard strains
Toxins of C. perfringens are responsible for enterotoxaemia in sheep [21]

Summary

Introduction

C. perfringens has a notorious reputation for possessing an extremely wide toxin arsenal, with over 15 toxins and may be the most widely occurring pathogenic in the globe [1, 2]. The organism is capable of producing different pathological pictures and of causing numerous different histotoxic and enteric diseases in both humans and other animals [1]. Alpha (α), beta (β), epsilon (ε) and iota (ι) comprise the four main C. perfringens toxins which form the basis of categorizing this bacterium into five different types, from A to E [3,4,5]. These different bacterial types provoke diverse diseases and even one strain may cause the infection in numerous hosts [6, 7]. In order to reduce or eliminate this risk, strategies must be developed to diagnose and prevent infected animals from entering the food chain [10]

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