Document 1120
 DOCN  M94A1120
 TI    Measurement of HIV antiviral activity by direct HIV RNA quantitation in
       frozen peripheral blood mononuclear cells (PBMCs) using a branched DNA
       (bDNA) assay.
 DT    9412
 AU    Anderson RE; Mohanty S; Wilber JC; Dailey PJ; Bristol-Myers Squibb Co.,
       Wallingford, CT.
 SO    Int Conf AIDS. 1994 Aug 7-12;10(2):204 (abstract no. PB0828). Unique
       Identifier : AIDSLINE ICA10/94371455
 AB    OBJECTIVE: To compare the infectious HIV viral titer with direct
       quantitation of HIV RNA in PBMCs. METHODS: Frozen separated PBMCs from
       subjects participating in a three dose study of stavudine (d4T) were
       titered for infectious HIV according to the ACTG method. HIV RNA in
       PBMCs was also quantitated by a nonisotopic DNA probe sandwich
       hybridization assay based on bDNA signal amplification (Quantiplex
       HIV-RNA assay, Chiron Corp.). PBMCs were homogenized in 8M guanidine HCL
       and 0.5% sarcosyl. RNA was precipitated with ethanol, resolubilized in
       buffer containing Proteinase K and detergent, and added directly to a
       96-well microplate for quantitation of HIV RNA. RESULTS: Median RNA
       levels were 6.2, 15.6, and 32.5 x 10(4) HIV equivalents per 10(7) cells
       and corresponded to 11, 52, and 256 Infectious Units/10(6) PBMCs in the
       lower, mid, and upper terciles of PBMC titers in all samples with
       detectable HIV RNA. HIV RNA decreased below baseline after 10 weeks in
       0/3, 4/5, and 2/3 subjects treated with 0.1, and 0.5 and 2.0 mg/kg/day
       of d4T respectively. CONCLUSION: The bDNA assay is a rapid technique
       which can be used to quantitate HIV RNA in PBMCs. This method may prove
       useful in examining dose effects and measuring activity of antiviral
       agents.
 DE    Comparative Study  DNA Probes  Human  HIV/GENETICS/*ISOLATION & PURIF
       HIV Infections/DRUG THERAPY/*MICROBIOLOGY  Leukocytes,
       Mononuclear/*MICROBIOLOGY  RNA, Viral/*ANALYSIS  Stavudine/*THERAPEUTIC
       USE  MEETING ABSTRACT

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