ENZYME-LINKED OLIGOSORBENT ASSAY FOR DETECTION OF POLYMERASE CHAIN REACTION-AMPLIFIED HUMAN-IMMUNODEFICIENCY-VIRUS TYPE-1

An enzyme-linked oligosorbent assay (ELOSA) was developed for the dete ction on microtiter plates of polymerase chain reaction (PCR)-amplifie d human immunodeficiency virus type 1 (HIV-1) DNA. The denatured PCR p roduct was hybridized with a passively adsorbed oligonucleotide captur e probe and a horseradish peroxidase-labeled oligonucleotide detection probe. The sensitivity and specificity of the PCR-ELOSA technique dep ended to some extent on the nucleotide sequences of the oligonucleotid e primer and probe quartet used in the amplification and detection. We evaluated five oligonucleotide quartets located in the gag, pol, vpr, env, and nef regions of HIV-1. DNAs from 39 HIV-1-seropositive indivi duals and 27 healthy HIV-1-seronegative controls were amplified by the PCR procedure, and the products were detected by ELOSA. Ten copies of HIV-1 DNA against a background of 1 mug of human DNA were specificall y detected by PCR-ELOSA. Specificities and sensitivities were, respect ively, 100 and 95% for the gag system, 100 and 97% for the pol system, 100 and 85% for the vpr system, 96 and 95% for the env system, and 10 0 and 95% for the nef system. The simplicity of ELOSA makes it suitabl e for automation and applicable to genetic testing and detection of vi ral and bacterial DNAs or RNAs in most routine laboratories.