Development of a m-PCR assay for simultaneous identification of Campylobacter jejuni and C. coli

TLDR

A multiplex PCR assay with three primer sets was developed to simultaneously identify Campylobacter jejuni and C. coli. The assay was applied to poultry faecal samples enriched in Preston broth, cultured on selective media, and tested by m‑PCR on bacterial isolates, with species identification confirmed by hippurate hydrolysis.

Abstract

Multiplex PCR assay (m–PCR) with three sets of primers was developed for simultaneous identification of Campylobacter jejuni and C. coli. Poultry faecal samples were enriched in Preston broth for 24 h and streaking on selective media was performed before and after enrichment. m–PCR was applied on bacterial cultures harvested from media plates. The data showed a selective effect of Preston broth which favoured the growth of C. coli. Identification of the species by the hippurate hydrolysis test and by the m–PCR was performed on 294 isolates of Campylobacter. The efficiency of the identification by the biochemical test is only 34% in comparison to 100% efficiency with the PCR. The use of our m–PCR in combination with the culture method allowed reliable detection and identification of C. jejuni and C. coli within 3–4 d.