本实验采用双抗体夹心ELISA法。抗HIV-1 Gag p24单抗包被于酶标板上,同时加入标本、标准品和生物素化的抗HIV-1 Gag p24抗体,标本、标准品中的HIV-1 Gag p24会与加入于孔中的生物素化的抗HIV-1 Gag p24抗体及包被于酶标板上的单抗结合,形成免疫复合物,游离的成分被洗去;加入辣根过氧化物酶标记的亲合素,亲合素与生物素特异性结合,游离的成分被洗去。加入显色底物(显色剂),若反应孔中有HIV-1 Gag p24,辣根过氧化物酶会使无色的显色剂变蓝,加终止液变黄。在450nm 处测OD值,HIV-1 Gag p24 浓度与OD450值之间呈正相关,可通过绘制标准曲线求出标本中HIV-1 Gag p24度。
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