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

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.