[Show abstract][Hide abstract] ABSTRACT: . A combined PCR-culture technique was developed to detect Arcobacter spp. in fresh chicken meat. Following a short selective enrichment of chicken samples, bacterial DNA was extracted and amplified using primers targeted at the genes encoding 16S rRNA of Arcobacter spp. The selected primers amplify a 181-bp fragment from all Arcobacter spp., whereas no PCR product is generated for other bacteria, including the closely related Campylobacter and Helicobacter species. The assay was used to screen 96 retail-purchased chicken samples for the presence of Arcobacter spp. Fifty-three percent of the samples analysed were positive for this micro-organism. The assay is simple and sensitive and reduces the amount of time required to positively detect Arcobacter spp. in poultry meat.
[Show abstract][Hide abstract] ABSTRACT: A rapid assay for enumeration of Arcobacter spp. in chicken meat was developed by using the polymerase chain reaction (PCR) coupled to an enzyme-linked immunosorbent assay (ELISA). Following a short selective enrichment of poultry samples, bacterial DNA was extracted and amplified using digoxigenin-labelled primers specific for 16S rRNA sequences of Arcobacter spp. Amplified fragments were heat denatured before being quantified by an ELISA. In this technique, a biotinylated probe immobilized onto streptavidin-coated microplates was used to capture the digoxigenin-PCR products. A peroxidase antidigoxigenin conjugate was added to the plate and, in the presence of substrate, PCR products were quantitated based on an optical density reading. Distinct absorbance differences were obtained when assaying poultry samples containing Arcobacter spp. in the range 10-10(4) CFU/g.