UTILIZATION OF A DNA ENZYME-IMMUNOASSAY FOR THE DETECTION OF PROVIRALDNA OF HUMAN-IMMUNODEFICIENCY-VIRUS TYPE-1 BY POLYMERASE CHAIN-REACTION

Background: The detection of proviral DNA by Polymerase Chain Reaction (PCR) is regarded as an important tool in the diagnosis of HIV-1 infe ction, specially among adults at risk of AIDS and children born to ser opositive mothers. However, application of PCR in routine testing is h ampered by the need to use radioactive probes. Objectives: In this stu dy, a non-radioactive test based on a microtiter plate (DNA Enzyme Imm unoAssay, DEIA) was used for the detection of proviral sequences of HI V-1 in peripheral blood cells of different patients. The results of th e PCR-DEIA assay were compared to those obtained by liquid hybridizati on (PCR-LH), virus isolation (VI) and Western blot (WB). Study design: The study population included 92 patients belonging to three differen t groups: seropositive subjects with a well-defined clinical status an d WB profile; adults at risk of infection with negative or indetermina te WB; children born to seropositive mothers with still unestablished HIV-1 infection. Results: In the seropositive subjects, both PCR-LH an d PCR-DEIA confirmed infection and gave the same results as WB. In adu lts at risk of infection, PCR with both methods anticipated the seroco nversion in one patient with indeterminate WB and confirmed the absenc e of infection among seronegative and other indeterminate patients. In children born to seropositive mothers, both PCR systems as well as VI permitted an early diagnosis of infection, as confirmed by the clinic al follow-up. Conclusion: This study has shown that in subjects at ris k of AIDS and in children born to seropositive mothers, the non-isotop ic DEIA method presents the same sensitivity and specificity for the d etection of HIV-1 infection as the radioactive procedure. The DEIA met hod appears to be particularly useful for the detection of PCR product s in routine diagnostic analyses.