J. Izopet et al., ASSESSMENT OF A STANDARDIZED REVERSE-TRANSCRIPTASE PCR ASSAY FOR QUANTIFYING HIV-1 RNA IN PLASMA AND SERUM, Journal of virological methods, 60(2), 1996, pp. 119-129
Citations number
30
Categorie Soggetti
Virology,"Biochemical Research Methods","Biothechnology & Applied Migrobiology
The analytical variability of the new commercially available Reverse T
ranscriptase-Polymerase Chain Reaction (RT-PCR) assay, Amplicor(TM) HI
V-I Monitor(TM), has been assessed to establish criteria for assessing
the significance of HIV-I RNA level measurements. Estimations of the
standard deviations (SD) of log-copies in inter-assay (mean 0.09 log)
and in inter-laboratory (mean 0.14 log) reproducibility experiments de
monstrated that the assay can discriminate with 95% confidence between
3-fold (inter-assay) and 5-fold differences (inter-laboratory). The i
nter-lot reproducibility (mean 0.10 log) was similar to the inter-assa
y reproducibility. The HIV-I RNA concentrations measured in plasma col
lected in potassium EDTA anticoagulant were slightly higher than those
measured in plasma collected in sodium citrate. The HIV-1 RNA concent
rations measured in sera were about 50%, of the HIV-1 RNA concentratio
ns measured in paired plasma samples. However, there was a strong corr
elation between these two measurements (P<0.0001). The assay was used
to measure viral RNA in the plasma of 50 HIV-1 positive individuals at
different stages of infection. All the individuals had detectable HIV
-I RNA (300-957 000 copies/ml). There was no correlation between HIV-I
RNA and Immune Complex Dissociated (ICD) p24 antigen, but HIV-I RNA w
as correlated with CD4+cell counts (P<0.0001) and the clinical stage (
P=0.0042), with higher HIV-1 RNA concentrations in patients with a mor
e advanced stage of the disease. The significant association of HIV-1
RNA with major markers of HIV infection and the reliability of this se
nsitive, easy-to-use RT-PCR assay indicate its suitability for use in
clinical trials and suggest that this assay is appropriate for routine
clinical applications.