       Document 0406
 DOCN  M9640406
 TI    Detection of HIV-1 infection in vitro using NASBA: an isothermal RNA
       amplification technique.
 DT    9604
 AU    Romano JW; Shurtliff RN; Sarngadharan MG; Pal R; Advanced BioSciences
       Laboratories Inc., Kensington, MD 20895,; USA.
 SO    J Virol Methods. 1995 Aug;54(2-3):109-19. Unique Identifier : AIDSLINE
       MED/96038350
 AB    Establishment of a sensitive infection assay for HIV-1 is essential for
       successful screening of antiviral agents and neutralizing antibodies. In
       this report, an infection assay is described which measures the
       expression of viral genomic RNA and spliced mRNA intermediates in
       infected cells by an amplification-based technique called NASBA. The
       extreme sensitivity of this method permits the detection of viral RNA in
       peripheral blood mononuclear cells (PBMC) within 48 h of infection by a
       low dose of virus. Similarly, spliced HIV-1 mRNA could be detected
       within 24 h of infection of CEM cells by HIV-1IIIB. This NASBA-based
       infection assay was shown to titer the neutralization of the HIV-1IIIB
       isolate by serum from an infected human and by a monoclonal antibody to
       gp120. Furthermore, the inhibitory effects of azidothymidine (AZT) and
       soluble CD4 on HIV-1IIIB infection were quantitated by this assay. The
       early detection of virus by NASBA minimizes the contribution of
       secondary infection, thereby permitting more accurate evaluation of
       antiviral agents and neutralizing antibodies. This assay may be useful
       for the study of infection of phenotypically distinct HIV-1 isolates,
       which differ in terms of their replication kinetics.
 DE    Antibodies, Viral/IMMUNOLOGY  Antigens, CD4/PHARMACOLOGY  Base Sequence
       Cells, Cultured  DNA Primers  Gene Amplification/*METHODS  Human
       HIV-1/DRUG EFFECTS/*GENETICS  Leukocytes, Mononuclear/*VIROLOGY
       Molecular Sequence Data  Neutralization Tests  RNA Splicing  RNA,
       Viral/*ANALYSIS  Zidovudine/PHARMACOLOGY  JOURNAL ARTICLE

       SOURCE: National Library of Medicine.  NOTICE: This material may be
       protected by Copyright Law (Title 17, U.S.Code).

