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    • 10. 发明申请
    • MULTIPLEX IMMUNO SCREENING ASSAY
    • 多重免疫屏蔽测定
    • WO2013083847A3
    • 2013-08-01
    • PCT/EP2012074986
    • 2012-12-10
    • PASTEUR INSTITUT
    • MANUGUERRA JEAN-CLAUDEVANHOMWEGEN JESSICADESPRES PHILIPPEPAULOUS SYLVIE
    • C12N9/10C12N15/62G01N33/543G01N33/564
    • G01N33/54306G01N33/54313G01N33/54353G01N33/564G01N33/56983G01N33/6845G01N2333/18G01N2333/185G01N2469/20
    • The present invention provides kits and assay methods for the early detection of pathogens, precise identification of the etiologic agent, and improved disease surveillance. More specifically, the present invention discloses an immunoassay leading to the rapid and simultaneous detection of antibodies to a wide range of infectious pathogens in biological fluids of infected patients. This immunoassay involves the covalent and oriented coupling of fusion proteins comprising an AGT enzyme and a viral antigen on an identifiable solid support (e.g. fluorescent microspheres), said support being previously coated with an AGT substrate. This coupling is mediated by the irreversible reaction of the AGT enzyme on its substrate. The thus obtained antigen-coupled microspheres show enhanced capture of specific antibodies as compared to antigen-coupled microspheres produced by standard amine coupling procedures. The methods of the invention possess the ability to multiplex, minimize the amount of biological sample, and have enhanced sensitivity and specificity toward target antibodies as compared with classical ELISA or Radio-Immunoprecipitation assays.
    • 本发明提供用于病原体的早期检测,病原体的精确鉴定和改进的疾病监测的试剂盒和测定方法。 更具体地,本发明公开了一种免疫测定,其导致快速和同时检测感染患者生物流体中广泛感染性病原体的抗体。 该免疫测定涉及包含AGT酶和病毒抗原的融合蛋白在可识别的固体支持物(例如荧光微球)上的共价和取向偶联,所述载体预先用AGT底物涂覆。 该偶联由AGT酶在其底物上的不可逆反应介导。 与通过标准胺偶联方法制备的抗原偶联微球相比,由此获得的抗原偶联微球显示出特异性抗体的增强捕获。 与传统的ELISA或放射免疫沉淀测定相比,本发明的方法具有复用多样化,最小化生物样品的量并且对靶抗体具有增强的敏感性和特异性的能力。