Reagent system for detecting HIV-infected peripheral blood lymphocytes in whole blood

摘要

Fluorometric immunological assay method for detection of HIV-1 infection in which Murine anti-gp120 monoclonal antibodies coupled to paramagnetic microspheres (14) and Fluorescein conjugated anti-gp120 polyclonal antibodies IgG (16) are incubated in a few drops of whole blood diluted in 0.5 cc phosphate buffered saline (10). After incubation for 5 minutes, the HIV-infected peripheral blood lymphocytes (18) will be coated with both the Murine anti-gp120 monoclonal antibodies coupled to paramagnetic microspheres (14) and Fluorescein conjugated anti-gp120 polyclonal antibodies IgG (16) at exposed gp120 antigens (20) binding sites. At the time of measurement said HIV- infected peripheral blood lymphocytes (18) will be pulled against the wall of the measurement vessel by means of a magnetic gradient (26). The cells adhering to the vessel wall are illuminated at 488 nm monochromatic light by a focused light source (28) and the resultant emitted fluorescence is imaged, measured and recorded.