• Title/Summary/Keyword: MESF(Molecules of Equivalent Soluble Fluorochrome)

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Quantitation of Antigen-Antibody Reaction Condition for Development of Fluorescence Image-based CD4 Rapid Test (형광 영상 기반 CD4 신속 검사법 개발을 위한 항원-항체 반응 조건 정량화)

  • Kim, Subin;Kim, Jung Kyung
    • Journal of the Korean Society of Visualization
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    • v.13 no.1
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    • pp.35-42
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    • 2015
  • 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.