Fast detection of Clostridium perfringens Type A,B,C and D by Multiplex PCR

Document Type : Full Research Paper

Authors

1 Biology Department, Basic Science Research Center, Imam Houssein University

2 Applied Biotechnology and Environmental Research Center, Baqiyatallah Medical Science University

3 Department of Anaerobiology, Razi Serum and Vaccine Institute, Karaj

Abstract

Clostridium perfringens is a part of the microflora of soil, and many reports have revealed the widespread occurrence of this organism in raw and processed foods. C. perfringens is classified into five types (types A to E), depending upon their ability to express alpha, beta, epsilon, and iota toxins In this study, a multiplex polymerase chain reaction (PCR) assay was developed for detection of C. perfringens types A, B, C and D. These strains have formerly been confirmed by the biochemical methods as well as serological test. For detection by PCR, four primer pairs with equal melting temperatures, were designed. DNA amplification fragments of 1167 bp for C. perfringens type A alone (α toxin), 1167 bp, 1025 bp and 961 bp for C. perfringens type B (α, β and ε toxin) 1167 bp and 1025 bp for C. perfringens type C (α and β toxin), and 1167 bp and 961 bp for C. perfringens type D (α and ε toxin) were obtained. Clostridium botulinum ued as negative control and did not yield a PCR product. These findings suggest that the assay is easy, timesaving, sensitive and specific and can use for detection of A, B, C and D types of C. perfringens.

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History

  • Receive Date: 26 April 2015
  • Accept Date: 26 April 2015

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