Document 0469
 DOCN  M9650469
 TI    Rapid culture and quantitation of human immunodeficiency virus type 1
       from patient cells without the use of mitogen-stimulated donor cells.
 DT    9605
 AU    Kim PK; He S; HO JL; Division of International Medicine, Cornell
       University Medical; College, New York, New York 10021, USA.
 SO    Clin Diagn Lab Immunol. 1994 Nov;1(6):660-6. Unique Identifier :
       AIDSLINE MED/96050887
 AB    We report the development of a rapid, sensitive virus culture method for
       direct quantitation of human immunodeficiency virus (HIV) in peripheral
       blood mononuclear cells (PBMCs). This new method involves culturing
       10(7) PBMCs from HIV-seropositive persons in 10 ml of medium containing
       phorbol 12-myristate 13-acetate and interleukin-2. Both agents stimulate
       cell activation and hence viral replication. Cell-associated virus and
       free virus are quantitated by a commercially available HIV p24 antigen
       capture enzyme immunoassay. Detection of cell-associated p24 antigen by
       flow cytometry was less sensitive than by the enzyme immunoassay. In
       this preliminary study, HIV was detected in 20 of 23 HIV-seropositive
       patients and in none of the 11 HIV seronegative low-risk individuals.
       One HIV-seronegative person with Guillain-Barre syndrome following
       high-risk activity was found to be rapid-HIV-culture positive. The
       overall sensitivity and specificity were 87 and 100%, respectively. By
       comparing the quantity of virus produced in infected cells with the
       amount of virus produced in chronically infected U1 monocytes and ACH-2
       lymphocytes stimulated with phorbol 12-myristate 13-acetate and
       interleukin-2, the approximate number of infected cells per sample is
       calculated. In the same patient specimens, quantitation of the number of
       HIV infected cells by the HIV rapid-culture method correlated with the
       results of the 21-day cell dilution coculture assay (correlation
       coefficient r = 0.5; 95% confidence interval, 0.07 to 0.77). Advantages
       of the rapid HIV culture include no requirement for donor PBMCs or
       change of media, shortened culture time, and the ability to detect p24
       viral antigen from cell-associated virus for quantitation of viral load.
 DE    Cell Count  Cell Line/VIROLOGY  Flow Cytometry  Human  HIV
       Antigens/BIOSYNTHESIS  HIV-1/*GROWTH & DEVELOPMENT/ISOLATION & PURIF
       Mitogens/PHARMACOLOGY  Monocytes/*VIROLOGY  Sensitivity and Specificity
       Support, Non-U.S. Gov't  Support, U.S. Gov't, P.H.S.  Time Factors
       JOURNAL ARTICLE

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