Cell culture supernatant
CDIA™ Lentivirus (HIV-1 P24) Semi-Quantitative Rapid Test Kit is a rapid chromatographic immunoassay for semi-quantitative detection of lentivirus titers in the supernatant of cell culture. This assay takes only 10-15 minutes, compared to the conventional re-infection based assay that takes 2-3 days.
Lentivirus vector based on the human immunodeficiency virus-1 (HIV-1) has become a promising vector for gene transfer studies. Lentiviral vectors have been shown to deliver genes toneurons, lymphocytes and macrophages, cell types that previous retrovirus vectors could not be used. Lentiviral vectors have also proven to be effective in transducing brain, liver, muscle, and retina in vivo without toxicity or immune responses. Lentivirus particles are produced from 293T cells through transient transfection of 3 or 4 plasmids that encodes for the components of the virion. Viral medium containing viral particles produced by packaging cells within 48-72 hr can be harvested. To ensure that pseudoviral medium is viable, and to control the number of copies of integrated viral constructs per target cell, the viral titer needs to be determined before proceeding with transduction experiments.
Principles of Testing
CDIA™ Lentivirus (HIV-1 P24) Semi-Quantitative Rapid Test Kit is a semi-quantitative, membrane based immunoassay for the detection of lentivirus titers. The membrane is pre-coated with anti-HIV-1 24 antibodies on the test line region. During testing, specimen reacts with the particles coated with anti-HIV-1 24 antibodies. The mixture migrates upward on the membrane by capillary action to react with anti- HIV-1 24 antibodies on the membrane and generate a colored line. The minimum detection level of this test is 1 ng/mL (T Line). When the HIV-1 P24 level in the specimen is between 1-500 ng/mL, it shows a positive correlation between the concentration of HIV-1 P24 in the specimen and the intensity of the color of the test line (T). The HIV-1 P24 concentration is semi-quantitatively determined by Color Chart (Fig 1), realizing the rapid estimation of lentivirus titer.
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