DNA Amplification for Direct Detection of HIV-1 in DNA of Peripheral Blood Mononuclear Cells

TLDR

The study proposes using PCR-based DNA amplification to detect HIV‑1 infection as an alternative to conventional virus isolation. The authors amplified proviral DNA from peripheral blood mononuclear cells using PCR with primers targeting multiple HIV‑1 genomic regions to maximize detection sensitivity. PCR identified HIV‑1 DNA in all seropositive samples with culture‑positive virus, in 64 % of culture‑negative seropositives, and in none of seronegatives, yielding results within 3 days versus weeks required for virus isolation.

Abstract

By means of a selective DNA amplification technique called polymerase chain reaction, proviral sequences of the human immunodeficiency virus (HIV-1) were identified directly in DNA isolated from peripheral blood mononuclear cells (PBMCs) of persons seropositive but not in DNA isolated from PBMCs of persons seronegative for the virus. Primer pairs from multiple regions of the HIV-1 genome were used to achieve maximum sensitivity of provirus detection. HIV-1 sequences were detected in 100% of DNA specimens from seropositive, homosexual men from whom the virus was isolated by coculture, but in none of the DNA specimens from a control group of seronegative, virus culture-negative persons. However, HIV-1 sequences were detected in 64% of DNA specimens from seropositive, virus culture-negative homosexual men. This method of DNA amplification made it possible to obtain results within 3 days, whereas virus isolation takes up to 3 to 4 weeks. The method may therefore be used to complement or replace virus isolation as a routine means of determining HIV-1 infection.