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

• Published in: Science (American Association for the Advancement of Science) Vol. 239; no. 4837; pp. 295 - 297
• Main Authors: Ou, Chin-Yih, Kwok, Shirley, Mitchell, Sheila W., Mack, David H., Sninsky, John J., Krebs, John W., Feorino, Paul, Warfield, Donna, Schochetman, Gerald
• Format: Journal Article
• Language: English
• Published: Washington, DC The American Association for the Advancement of Science 15.01.1988; American Association for the Advancement of Science
• Subjects: Acquired immune deficiency syndrome; Acquired Immunodeficiency Syndrome - microbiology; AIDS; AIDS related complex; AIDS/HIV; Base Sequence; Biochemistry; Biological and medical sciences; Cultural groups; DNA; DNA polymerases; DNA probes; DNA, Viral - blood; DNA-Directed DNA Polymerase; Fundamental and applied biological sciences. Psychology; Gene Amplification; Genetics; HIV; HIV (Viruses); HIV - genetics; HIV - isolation & purification; HIV 1; HIV Seropositivity; Homosexuality; Human immunodeficiency virus 1; Humans; Leukocytes, Mononuclear - analysis; Lymphocytes; Male; Medical research; Microbiology; Monocytes; Nucleic Acid Amplification Techniques; Nucleic Acid Hybridization; Peripheral blood mononuclear cells; Polymerase chain reaction; Proviruses; Serodiagnosis; Serologic tests; Techniques used in virology; Testing; Virology; Virus Cultivation; Viruses; Human; Antibody; Retroviridae; Method; Infection; Virus; Polymerase chain reaction; Gene amplification; Mononuclear cell; DNA; Viral disease; Séropositivity; Microorganism culture; Diagnosis; Human immunodeficiency virus; Detection; Comparative study
• ISSN: 0036-8075; 1095-9203
• DOI: 10.1126/science.3336784

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.