Comparative restriction enzyme mapping of Campylobacter jejuni isolates from turkeys and broilers based on flaA flagellar gene using HpyF3I endonuclease

Abstract

Turkeys and broilers have been identified as important reservoirs for Campylobacter jejuni which is of public health significance. The evaluation of the genotypes among C. jejuni strains within different reservoirs is critical for our understanding of the epidemiology of this infectious agent. The present study aimed to compare the genetic diversity and differences of C. jejuni isolates from turkeys and broilers using flagellin PCR-RFLP typing (flaA typing) technique, in terms of the ease of use and discriminatory power. Sixty C. jejuni isolates were detected biochemically and confirmed by duplex-PCR from turkeys and broilers (30 strains from each bird species). Then, a flaA gene fragment (1725 bp) of C. jejuni isolates was amplified and amplicons were digested with HpyF3I enzyme. Restriction analysis by HpyF3I gave four different flaA patterns (H1, H2, H3, H4) among all tested C. jejuni isolates. In broiler isolates, all four patterns were observed but in turkey isolates, only H2 and H4 patterns were present. The results clearly demonstrated that distribution of the flaA typing patterns differed depending on the host species (broiler/turkey). H1 and H3 flaA types are more prevalent in broiler than turkey isolates, while H2 type is significantly more prevalent within isolates from turkey (p < 0.05). The flaA typing technique by digestion with HpyF3I enzyme can almost give us a clue to the source of infection in local outbreaks.

Research Progresses on the Function and Detection Methods of Insect Gut Microbes

“…Both RFLP and ARDRA require the use of endonucleases to digest specific PCR amplification products and then obtain fragments of different numbers and lengths to distinguish bacteria. This method is suitable for strains that have been successfully isolated and can determine whether there are differences in the genus of a large number of strains [ 90 , 91 ] ( Table 2 ). He et al identified 12 and 11 species in the Camponotus midgut, respectively, utilizing traditional culture methods and 16S rRNA-RFLP technology [ 92 ].…”