DNA amplification for direct detection of HIV-1 in DNA of peripheral blood mononuclear cells.
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.
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Center for Infectious Diseases, Centers for Disease Control, Atlanta, GA 30333.
Kwok
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Mitchell
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Mack
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Sninsky
J J
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Krebs
J W
JW
Feorino
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Warfield
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Schochetman
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G
D000163:Acquired Immunodeficiency Syndrome / Q000382:microbiology*; D001483:Base Sequence; D004279:DNA, Viral / Q000097:blood*; D004259:DNA-Directed DNA Polymerase; D005784:Gene Amplification; D006678:HIV / Q000235:genetics* / Q000302:isolation & purification; D006679:HIV Seropositivity; D006716:Homosexuality; D006801:Humans; D007963:Leukocytes, Mononuclear / Q000032:analysis*; D008297:Male; D021141:Nucleic Acid Amplification Techniques; D009693:Nucleic Acid Hybridization; D014776:Virus Cultivation
D004279:DNA, Viral; D004259:DNA-Directed DNA Polymerase