Quantitation of HIV viral burden by PCR in HIV seropositive Navy personnel representing Walter Reed stages 1 to 6.
To quantitate the amount of human immunodeficiency virus type 1 (HIV-1) DNA in peripheral blood mononuclear cells (PBMC) of 78 infected individuals, we have developed a polymerase chain reaction (PCR) assay that is both quantitative and sensitive. Quantitation was based on incorporation of a 32P end-labelled primer (SK39) in the PCR reaction and on comparison after electrophoresis with known amounts of HIV DNA. A linear relationship was obtained between the natural logarithms of the radioactive counts detected and the number of HIV-1 DNA copies (10-1000 copies) from the standard DNA. HIV copy numbers from patient samples were then extrapolated from the standard curves. This sensitive and reproducible method was compared with virus isolation which is a semiquantitative evaluation of viral burden. HIV DNA levels correlated with virus isolation, i.e., high viral burden (100-1000 HIV copies) were found in most samples from which virus was isolated after only 7 days in culture; low viral burden (less than 100 HIV copies) was observed in samples from which virus was isolated after 14 to 21 days in culture. These estimates of viral burden were then compared with the clinical stage of the individuals.
Ferre
F
F
Immune Response Corporation, Carlsbad, CA 92008.
Marchese
A
A
Duffy
P C
PC
Lewis
D E
DE
Wallace
M R
MR
Beecham
H J
HJ
Burnett
K G
KG
Jensen
F C
FC
Carlo
D J
DJ
D004279:DNA, Viral / Q000032:analysis*; D006679:HIV Seropositivity / Q000382:microbiology* / Q000473:pathology; D015497:HIV-1 / Q000302:isolation & purification*; D006772:Hospitals, Military; D006801:Humans; D007958:Leukocyte Count; D008889:Military Personnel; D016133:Polymerase Chain Reaction / Q000379:methods*; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity
D004279:DNA, Viral