Submitted: 23 Apr 2017
Revised: 01 Jul 2017
Accepted: 08 Aug 2017
First published online: 31 Aug 2017
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Avicenna J Clin Microbiol Infect. 2017;4(3): e11983.
doi: 10.5812/ajcmi.11983
  Abstract View: 81
  PDF Download: 295

Research Article

Polymerase Chain Reaction-Restriction Fragment Length Polymorphism (PCR-RFLP) Analysis of Three Lipooligosaccharide-Associated Genes of Campylobacter jejuni and Campylobacter coli Isolated From Animal Samples

Rahem Khoshbakht 1 * , Mohammad Tabatabaei 2, Saeid Hosseinzadeh 3

1 Department of Food Hygiene, Faculty of Veterinary Medicine, Amol University of Special Modern Technologies, Amol, IR Iran
2 Department of Pathobiology, School of Veterinary Medicine, Shiraz University, Shiraz, IR Iran
3 Department of Public Health and Food Hygiene, School of Veterinary Medicine, Shiraz University, Shiraz, IR Iran
Corresponding author: Rahem Khoshbakht, DVM, Ph.D in Bacteriology, Department of Pathobiology, Faculty of Veterinary Medicine, Amol University of Special Modern Technologies, Emam Khomeini street, Aftab 24, Amol, Iran. Tel: +98-9113919059, Fax: +98-1144271054 Email: khoshbakht.r@gmail.com

Article

Background: Several genetic mechanisms are used by Campylobacter spp. to achieve pathogenesis. One of the involved virulence factors is lipooligosacharide-associated genes, which are related to ganglioside mimicry by Campylobacter species.

Objectives: The current study was conducted to determine the genetic diversity of 3 LOS-associated genes among Campylobacter jejuni and C. coli isolated from animal fecal samples.

Methods: One hundred broiler, cattle, and sheep fecal C. jejuni and C. coli isolates, which had been collected previously from Shiraz slaughterhouses and had been formerly identified by polymerase chain reaction (PCR) reactions, were used in the present study. Campylobacter species were subjected to detect wlaN, cgtB, and waaC genes. The PCR products of three LOS-genes were subjected for restriction fragment length polymorphism (RFLP) using HindIII and AluI restriction enzymes in separate reactions. The most prevalent RFLP patterns in the combination of 2 enzymes were subjected for sequencing and sequence analysis software.

Results and Conclusions: Patterns of RFLP for PCR products of all 69 wlaN and 29 cgtB genes were similar yet among 86 waaC gene PCR products, 6 different RFLP patterns were obtained. In conclusion, PCR-RFLP analysis demonstrated considerable variation in gene content and overall sequence heterogeneity in the animal-associated C. jejuni and C. coli LOS biosynthesis genes.

Copyright © 2017, Avicenna Journal of Clinical Microbiology and Infection. This is an open-access article distributed under the terms of the Creative Commons Attribution-NonCommercial 4.0 International License (http://creativecommons.org/licenses/by-nc/4.0/) which permits copy and redistribute the material just in noncommercial usages, provided the original work is properly cited.
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