Document 2745
 DOCN  M94A2745
 TI    Quantitation of serum HIV-RNA Using MRT-PCR.
 DT    9412
 AU    Tsukada K; Katayama K; Meguro T; Taki M; Itoh H; Yamada K; Oya A; Basic
       Research Div., BML Inc., Saitama, Japan.
 SO    Int Conf AIDS. 1994 Aug 7-12;10(1):234 (abstract no. PB0365). Unique
       Identifier : AIDSLINE ICA10/94369830
 AB    OBJECTIVE: Immunological and virological methods provide useful
       information as predictors of AIDS development in HIV infected patients.
       Multycyclic Reverse Transcriptase-Polymerase Chain Reaction (MRT-PCR)
       was established to quantitate serum HIV-RNA from HIV infected patients.
       Quantitative analysis of HIV-RNA from patient sera was performed in
       attempt to predict the prognosis of HIV carriers. MATERIALS AND METHODS:
       Serially collected samples from hemophiliac patients and homosexual
       patients with HIV infection (AC to AIDS) were analyzed retrospectively.
       HIV-RNA was purified from 100 microliters patient sera by AGPC method.
       Standard HIV-RNA was made from SP6 promoted HIV gag construct. MRT-PCR
       was performed using purified patient RNA and standard RNA samples with
       primers for gag. Aliquots of PCR products taken at 5 cycle intervals
       were subjected to dot-hybridization. HIV-RNA levels were determined by
       comparing the intensity of the patient signal to the standard RNA signal
       on the autoradiogram. RESULTS: Lineality was obtained at 10(4)-10(9)
       copies/ml at 30 cycles and 10(0)-10(5) copies/ml at 45 cycles. By using
       30 and 45 cycles, quantitation of HIV-RNA was possible from 10(0)
       copies/ml to 10(9) copies/ml. HIV-RNA was detected from all patient
       samples. In AC group, the amount of HIV-RNA was 10(3)-10(5) copies/ml.
       In AIDS/ARC group, the amount of HIV-RNA was 10(5)-10(7) copies/ml.
       DISCUSSION: The results from MRT-PCR were compatible with the CRT-PCR
       while the methodology was simpler and required less specimen volume.
       HIV-RNA levels reflected the clinical stage of the patient. Quantitation
       of HIV-RNA in serum may be useful as marker for patient prognosis and
       monitoring drug response.
 DE    Acquired Immunodeficiency Syndrome/DIAGNOSIS/MICROBIOLOGY
       Autoradiography  AIDS-Related Complex/DIAGNOSIS/MICROBIOLOGY
       Hemophilia/BLOOD  Homosexuality  Human  HIV/*GENETICS  HIV
       Infections/*DIAGNOSIS/MICROBIOLOGY  Male  Polymerase Chain
       Reaction/*METHODS  Predictive Value of Tests  Retrospective Studies
       RNA, Viral/*BLOOD  MEETING ABSTRACT

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