Document 0137
 DOCN  M9440137
 TI    Ultrasensitive retrovirus detection by a reverse transcriptase assay
       based on product enhancement.
 DT    9404
 AU    Pyra H; Boni J; Schupbach J; Swiss National Center for Retroviruses,
       Institute of Medical; Virology, University of Zurich.
 SO    Proc Natl Acad Sci U S A. 1994 Feb 15;91(4):1544-8. Unique Identifier :
       AIDSLINE MED/94151366
 AB    Reverse transcriptase (RT) is an indispensable component of infectious
       retroviruses. We have developed an ultrasensitive RT test in which RNA
       of bacteriophage MS2 serves as the template for RT-mediated cDNA
       synthesis. A fragment of the cDNA is selectively amplified by polymerase
       chain reaction and the amplification product is analyzed by Southern
       blot hybridization or enzyme immunoassay. The procedure was 10(6) to
       10(7) times more sensitive than a conventional RT test and detected as
       little as 10(-9) unit of murine leukemia virus RT, which corresponded to
       2.1 x 10(2) molecules, a number present in 3-11 virions. As a screening
       assay for filterable particle-associated RT, it was positive with
       supernatants from cell cultures producing human immunodeficiency virus
       (HIV) type 1 or human T-cell leukemia virus (HTLV) type 1 or 2, but was
       negative with nonproducer cultures. It was positive with plasma samples
       from all tested individuals infected with HIV-1, HIV-2, or HTLV-1 and
       sera from cats infected with feline leukemia virus or feline
       immunodeficiency virus. Control samples from blood donors or uninfected
       cats were negative. Density banding experiments with culture
       supernatants showed that the RT activity was associated with virus
       particles. The assay should detect all replication-competent
       retroviruses or similar agents. It may be used as a screening assay for
       such agents, for quantitation of the viral load, drug susceptibility
       testing of RT, and control of virus inactivation in biological products.
 DE    Animal  Base Sequence  Cats  Feline Acquired Immunodeficiency
       Syndrome/BLOOD/ENZYMOLOGY  Human  HIV Infections/BLOOD/ENZYMOLOGY
       HIV-1/ENZYMOLOGY/ISOLATION & PURIF  HTLV Infections/BLOOD/ENZYMOLOGY
       Leukemia Viruses, Murine/ENZYMOLOGY  Leukocytes,
       Mononuclear/MICROBIOLOGY  Molecular Sequence Data  Polymerase Chain
       Reaction  Retroviridae/*ENZYMOLOGY/ISOLATION & PURIF  Retroviridae
       Infections/*BLOOD  Reverse Transcriptase/*ANALYSIS  Sensitivity and
       Specificity  Support, Non-U.S. Gov't  JOURNAL ARTICLE

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       protected by Copyright Law (Title 17, U.S.Code).