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

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