A rapid method for quantitating the frequency of peripheral blood cells containing HIV-1 DNA.
This article describes a novel approach to quantitative polymerase chain reactions (PCR). The technique is simple to execute, can be performed in a single tube, and is suitable for automation. In addition, the counting and lysis of low numbers of cells (1-100) can be confirmed by phase contrast microscopy. In this study, the technique was used to determine the frequency of occurrence of DNA from the human immunodeficiency virus (type 1) in leukocyte subsets of peripheral blood mononuclear cells (PBMC) from sero-positive individuals. It was confirmed that the frequency of infected CD4+ T cells varied from 0.01 to 0.0001. In addition, HIV-1 DNA was detected in the B cell/dendritic cell-enriched subpopulation in four of nine HIV-1-positive individuals in the study.
Poznansky
M C
MC
Dana Farber Cancer Institute, Department of Pathology, Harvard Medical School, Boston, MA 02115.
Walker
B
B
Haseltine
W A
WA
Sodroski
J
J
Langhoff
E
E
D000386:AIDS-Related Complex / Q000097:blood / Q000382:microbiology; D000163:Acquired Immunodeficiency Syndrome / Q000097:blood / Q000382:microbiology*; D001402:B-Lymphocytes / Q000382:microbiology; D001483:Base Sequence; D015496:CD4-Positive T-Lymphocytes / Q000382:microbiology; D004279:DNA, Viral / Q000032:analysis* / Q000737:chemistry; D003713:Dendritic Cells / Q000382:microbiology; D015658:HIV Infections / Q000097:blood / Q000382:microbiology*; D006679:HIV Seropositivity / Q000097:blood / Q000382:microbiology; D015497:HIV-1 / Q000235:genetics / Q000302:isolation & purification*; D006801:Humans; D007958:Leukocyte Count; D007963:Leukocytes, Mononuclear / Q000382:microbiology*; D008297:Male; D008969:Molecular Sequence Data; D009841:Oligonucleotides / Q000737:chemistry; D016133:Polymerase Chain Reaction
D004279:DNA, Viral; D009841:Oligonucleotides