Since detection of amplified polymerase chain reaction products is a complicated procedure, a colorimetric microtiter plate hybridization assay was developed to automate specific detection of amplified HIV targets. In this assay, hybridization is seen by an antibody reacting selectively with double-stranded DNA. One hundred and ten amplified products detected by a DNA enzyme immunoassay (DEIA) and by classical hybridization with a digoxigenin-labeling probe, detection sensitivities were less than 10 HIV targets per 10(6) cells. This study demonstrates the specificity and sensitivity of DEIA for detecting amplified HIV targets. The use of the same equipment as for ELISA and the complete automation of the procedure allow a large number of samples to be processed in the clinical laboratory.