AUTOMATED DETECTION OF DIGOXIGENIN-LABELED B19 PARVOVIRUS AMPLICONS BY A CAPTURE HYBRIDIZATION ASSAY

An automated method to identify B19 amplicons, directly labelled with digoxigenin during amplification reaction was developed. The labelled amplicons were hybridized with a biotinylated B19 oligo-probe and capt ured on commercially available test tubes coated with streptavidin. Th e hybridized amplicons labelled with digoxigenin were detected using a nti-digoxigenin Fab fragments conjugated to peroxidase and the colouri metric reaction automatically evaluated as an immunoenzymatic assay. F ifty serum samples were tested by the assay and the results were in ac cordance with those obtained by Southern blot analysis of amplified pr oducts. Due to the high sensitivity, specificity and reproducibility s hown, the assay seems to be a practical and reliable test for the diag nosis of B19 infection and can be easily adapted to identify any digox igenin-labelled amplified product of viral genomes.