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http://dx.doi.org/10.5407/jksv.2015.13.1.035

Quantitation of Antigen-Antibody Reaction Condition for Development of Fluorescence Image-based CD4 Rapid Test  

Kim, Subin (국민대학교 기계공학과)
Kim, Jung Kyung (국민대학교 기계공학과)
Publication Information
Journal of the Korean Society of Visualization / v.13, no.1, 2015 , pp. 35-42 More about this Journal
Abstract
CD4+ T-cell count determines the effectiveness for antiretroviral therapy (ART) in patients with human immunodeficiency virus (HIV). Although ART slows the progression of HIV to AIDS, rapid counting of CD4+ T lymphocytes with a drop of patient's blood sample is urgently needed to ensure timely ART treatment in rural areas. Recently point-of-care CD4 testing devices have been developed by using non-flow based imaging cytometer incorporated with a sample cartridge where CD4+ T cells are reacted with fluorescently tagged specific antibodies. Here we conducted an experimental study using a conventional fluorescence microscope-based imaging system to quantitate the interaction of CD4 antibodies with CD4+ T cells at different reaction conditions. We demonstrated that a fast and affordable point-of-care CD4 test is feasible with a far less amount of antibodies and a shorter incubation time compared with a conventional sample preparation protocol for flow cytometry. We also proposed a general method to evaluate and compare the detection limit across different CD4 counting platforms by using fluorescently labelled microbeads for intensity calibration.
Keywords
CD4+ T lymphocyte; Fluorescence; Antigen-Antibody Reaction; Point-of-Care Test; Calibration Bead; MESF(Molecules of Equivalent Soluble Fluorochrome);
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