Application of Triplex PCR Technique in Identification of Clostridium perfringens B, C and D Types

Document Type : Scientific - Research Articles

Authors

mohammad reza Ahsani ¹

Mohammadreza Mohammad Abadi ²

mehrdad Shamsodini bafti ³

majed Ezatkhah ³

Mahdei Hasani Derakhshan ³

ali Esmailzadeh koshkooeh ²

¹ Department of Animal Genetics and Breeding, Department of Animal Science, Shahid Bahonar University of Kerman, Kerman, Iran

² Department of Animal Science, Faculty of Agriculture, Shahid Bahonar University of Kerman, Kerman, Iran

³ Razi Vaccine and Serum Research Institute, Southeast Branch, Kerman, Iran

10.22067/ijasr.v2i2.4106

Abstract

In this research 130 sheep-dung samples were collected from Kermani breed in area of Kerman randomly. After processing
and culture of samples, the produced colonies were studied morphologically and gram stain test and the genus of these
bacteria were identified using biochemical tests. The Clostridia strains isolates were included 30 percent of total samples i.e.
clostridiums perfringense, baratii, absonum, bifermentans, sporogenes, leptum, aurantibutyricum, sporosphaeroides,
symbiosum, scatologenes, ramosum and sordellii. DNA extracted from the isolated bacteria. For genotyping of 39 bacteria,
the extracted DNA was tested by triplex PCR. Based on length of synthesised fragment in PCR, different types of toxin and
strains of this bacterium were detected that from 23 samples of clostridium perfringense isolated using biochemical tests, all
samples shown 324 bp fragment. From these 5 samples were detected as B type, 8 samples as C type and 6 samples as D
type.

Keywords

Clostridium perfringens

Biochemical testes

DNA

Triplex PCR

---