CHEMILUMINESCENT DETECTION OF AMPLIFIED PSEUDORABIES VIRUS GP50 DNA WITH IMMOBILIZED PROBES ON MICROTITER WELLS

A highly specific and sensitive PCR assay for the envelope glycoprotei n gp50 has been developed for the detection of PRV-DNA sequences. Prim er pairs from PRV gp50 gene were used with the enzyme uracil N-glycosy lase and dUTP instead dTTP to prevent contamination due to PCR product carry-over. Biotinylated PCR products were captured in microtiter wel ls by specific oligonucleotide covalently linked to the polystyrene we lls. After recognition of the biotinylated PCR product with a streptav idin phosphatase conjugate, a chemiluminescent detection was realised. Different factors influencing the binding, the hybridization and the detection efficiency have been tested.