Enzyme-linked oligosorbent assay for detection of polymerase chain reaction-amplified human immunodeficiency virus type 1.

摘要: An enzyme-linked oligosorbent assay (ELOSA) was developed for the detection on microtiter plates of polymerase chain reaction (PCR)-amplified human immunodeficiency virus type 1 (HIV-1) DNA. The denatured PCR product hybridized with a passively adsorbed oligonucleotide capture probe and horseradish peroxidase-labeled probe. sensitivity specificity PCR-ELOSA technique depended to some extent nucleotide sequences primer quartet used in amplification detection. We evaluated five quartets located gag, pol, vpr, env, nef regions HIV-1. DNAs from 39 HIV-1-seropositive individuals 27 healthy HIV-1-seronegative controls were amplified by procedure, products detected ELOSA. Ten copies HIV-1 DNA against background microgram specifically PCR-ELOSA. Specificities sensitivities were, respectively, 100 95% gag system, 97% pol 85% vpr 96 env system. simplicity ELOSA makes it suitable automation applicable genetic testing viral bacterial or RNAs most routine laboratories.