Detection of human immunodeficiency virus type 1 in AIDS patients using amplification-mediated hybridization analyses: reproducibility and quantitative limitations.
Eighty-six peripheral blood mononuclear cell (PBMC) samples from 30 patients with AIDS were analyzed using a transcription-based amplification system (TAS) and the polymerase chain reaction (PCR). Human immunodeficiency virus type 1 (HIV-1) sequences were detected by amplification-mediated hybridization in 98% of the samples, 52% of which were positive for p24 antigen by ELISA. Neither TAS (93%) nor PCR (95%) detected HIV-1 sequences in all 86 samples. The hybridization-detection methods (slot blot, bead-based sandwich, and solution) used to detect the HIV-1-specific TAS products had a clear influence on the efficiency of detecting and quantitating the levels of HIV-1 present in these samples. The reproducibility of amplification of constant amounts of HIV-1 RNA and beta-globin DNA by TAS and PCR was studied over 3 months. The results indicated that variations of 10- and 5-fold in the HIV-1 sequence levels could be detected between samples by TAS and PCR, respectively. Within the range of sensitivities for each assay used, the administration of zidovudine did not appear to reduce the amount of HIV-1 nucleic acid sequences as observed in PBMC obtained serially from six AIDS patients.
Davis
G R
GR
Salk Institute Biotechnology/Industrial Associates, San Diego, California.
Blumeyer
K
K
DiMichele
L J
LJ
Whitfield
K M
KM
Chappelle
H
H
Riggs
N
N
Ghosh
S S
SS
Kao
P M
PM
Fahy
E
E
Kwoh
D Y
DY
D000163:Acquired Immunodeficiency Syndrome / Q000382:microbiology*; D001483:Base Sequence; D005784:Gene Amplification; D015683:Gene Products, gag / Q000032:analysis; D015488:HIV Antigens / Q000032:analysis; D016655:HIV Core Protein p24; D015497:HIV-1 / Q000235:genetics / Q000302:isolation & purification*; D006801:Humans; D008969:Molecular Sequence Data; D009693:Nucleic Acid Hybridization; D016133:Polymerase Chain Reaction; D011237:Predictive Value of Tests; D015203:Reproducibility of Results; D014158:Transcription, Genetic; D014758:Viral Core Proteins / Q000032:analysis
D015683:Gene Products, gag; D015488:HIV Antigens; D016655:HIV Core Protein p24; D014758:Viral Core Proteins