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

A rapid assay for enumeration of Arcobacter spp. in chicken meat was develo ped by using the polymerase chain reaction (PCR) coupled to an enzyme-linke d immunosorbent assay (ELISA). Following a short selective enrichment of po ultry samples, bacterial DNA was extracted and amplified using digoxigenin- labelled primers specific for 16S RNA sequences of Arcobacter spp. Amplifie d fragments were heat denatured before being quantified by an ELISA. In thi s technique, a biotinylated probe immobilized onto streptavidin-coated micr oplates was used to capture the digoxigenin-PCR products. A peroxidase anti digoxigenin conjugate was added to the plate and, in the presence of substr ate, PCR products were quantitated based on an optical density reading. Dis tinct absorbance differences were obtained when assaying poultry samples co ntaining Arcobacter spp. in the range 10-10(4) cfu/g. (C) 2001 Elsevier Sci ence Ltd. All rights reserved.