Arcobacter spp. enumeration in poultry meat using a combined PCR-ELISA assay

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 RNA 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. (C) 2001 Elsevier Science Ltd. All rights reserved.