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In this study, we described an ultrasensitive and high-throughput luciferase immunosorbent assay (LISA) for qualitative and quantitative detection of anti-HIV-1 antibody. Anti-HIV antibody in serum or plasma samples was captured by protein A/G-coated microtiter plate and detected with crude cell lysates expressing Nanoluc luciferase (Nluc) enzyme fused with HIV-1 p24 or gp41 antigen without the need of protein purification. After the addition of furimazine substrate, anti-HIV antibodies were quantitatively measured as luciferase light units. LISA showed a wide linear range of detection and was about 10-fold more sensitive than ELISA. For the detection of both anti-p24 and anti-gp41, LISA showed extraordinary sensitivity (99.5% and 100%, respectively) and equivalent specificity (100%). LISA could also monitor the change in the anti-HIV-1 antibody response over time in antiretroviral therapy (ART) treated individuals, and can sufficiently distinguish between recent and long-term HIV-1 infections. Our preliminary results indicate that LISA may provide a novel universal immunoassay platform for simultaneous HIV-1 detection, quantitative measurement of anti-HIV antibodies as well as the differentiation of HIV-1 infection stages.
This article was published in the following journal.
Name: Virus research
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Rapid methods of measuring the effects of an agent in a biological or chemical assay. The assay usually involves some form of automation or a way to conduct multiple assays at the same time using sample arrays.
An immunoassay utilizing an antibody labeled with an enzyme marker such as horseradish peroxidase. While either the enzyme or the antibody is bound to an immunosorbent substrate, they both retain their biologic activity; the change in enzyme activity as a result of the enzyme-antibody-antigen reaction is proportional to the concentration of the antigen and can be measured spectrophotometrically or with the naked eye. Many variations of the method have been developed.
The assay of INTERFERON-GAMMA released from lymphocytes after their exposure to a specific test antigen, to check for IMMUNOLOGIC MEMORY resulting from a previous exposure to the antigen. The amount of interferon-gamma released is usually assayed by an ENZYME-LINKED IMMUNOSORBENT ASSAY.
A method of detection of the number of cells in a sample secreting a specific molecule. Wtih this method a population of cells are plated over top of the immunosorbent substrate that captures the secreted molecules.
Techniques for removal by adsorption and subsequent elution of a specific antibody or antigen using an immunosorbent containing the homologous antigen or antibody.
An assay is an analytic procedure for qualitatively assessing or quantitatively measuring the presence or amount or the functional activity of a target entity. This can be a drug or biochemical substance or a cell in an organism or organic sample. ...
AIDS and HIV
AIDS; Acquired Immune Deficiency Syndrome. HIV; Human Immunodeficiency Virus HIV infection causes AIDS. HIV infection also causes the production of anti-HIV antibodies, which forms the test for HIV in patients. People who have the HIV antibodies are ...
An antibody is a protein produced by the body's immune system when it detects harmful substances, called antigens. Examples of antigens include microorganisms (such as bacteria, fungi, parasites, and viruses) and chemicals. Antibodies may be produc...