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M94A2748.TXT
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1994-10-25
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Document 2748
DOCN M94A2748
TI Use of a quantitative PCR assay for measurement of HIV RNA in plasma
during infection and seroconversion.
DT 9412
AU Herman S; Frenkl T; Mulder J; Payne H; Wang Z; Spadoro J; Roche
Molecular Systems, Branchburg, NJ.
SO Int Conf AIDS. 1994 Aug 7-12;10(1):233 (abstract no. PB0362). Unique
Identifier : AIDSLINE ICA10/94369827
AB OBJECTIVE: Use of a Polymerase Chain Reaction (PCR)-based assay for
quantitative determination of HIV RNA in plasma specimens during
seroconversion, and comparison to Ab and Ag titers, as determined with
commercially available test kits. METHODS: A PCR-based assay for
quantitation of HIV-1 RNA in plasma has been developed. The assay uses a
simplified amplification procedure in which reverse transcription of
viral RNA and PCR are carried out in a single reaction by one enzyme.
PCR products are detected by hybridization to oligonucleotide probes in
a microwell plate format, yielding a colorimetric result. Quantitation
is achieved by comparison to an internal RNA standard added to every
sample during sample processing. The assay has an analytical sensitivity
of 10 HIV RNA molecules, and a dynamic range of greater than 3 orders of
magnitude. The assay was used for quantitation of HIV RNA in plasma
units collected serially from individual donors during seroconversion,
and the results were compared to Ab and Ag titers (specimens, Ab and Ag
results kindly provided by Boston Biomedica, Inc., W. Bridgewater, MA,
USA). RESULTS: In all 7 donors an acute phase of viral infection was
noted by high HIV RNA titers, followed by suppression of RNA titers by 2
to 3 orders of magnitude over periods of 7 to 35 days, coincident with
seroconversion. Although the RNA and Ag titers followed the same general
patterns, HIV RNA remained detectable by the PCR test after
seroconversion in all 7 donors, whereas Ag became undetectable in 6 of
the 7 donors. CONCLUSIONS: The quantitative PCR assay detected the acute
phase of viral infection, and was sufficiently sensitive to detect viral
RNA after seroconversion, when Ag was undetectable, suggesting that the
assay may be useful for monitoring viral load in patients with low and
high viral burdens.
DE AIDS Serodiagnosis Human HIV Antibodies/BLOOD HIV Antigens/BLOOD HIV
Infections/*DIAGNOSIS/MICROBIOLOGY HIV
Seropositivity/*DIAGNOSIS/MICROBIOLOGY HIV-1/*GENETICS Polymerase
Chain Reaction/*METHODS Predictive Value of Tests RNA, Viral/*BLOOD
MEETING ABSTRACT
SOURCE: National Library of Medicine. NOTICE: This material may be
protected by Copyright Law (Title 17, U.S.Code).