本實(shí)驗(yàn)采用雙抗體夾心ELISA法。抗HIV-1 Gag p24單抗包被于酶標(biāo)板上,同時(shí)加入標(biāo)本、標(biāo)準(zhǔn)品和生物素化的抗HIV-1 Gag p24抗體,標(biāo)本、標(biāo)準(zhǔn)品中的HIV-1 Gag p24會(huì)與加入于孔中的生物素化的抗HIV-1 Gag p24抗體及包被于酶標(biāo)板上的單抗結(jié)合,形成免疫復(fù)合物,游離的成分被洗去;加入辣根過氧化物酶標(biāo)記的親合素,親合素與生物素特異性結(jié)合,游離的成分被洗去。加入顯色底物(顯色劑),若反應(yīng)孔中有HIV-1 Gag p24,辣根過氧化物酶會(huì)使無色的顯色劑變藍(lán),加終止液變黃。在450nm 處測OD值,HIV-1 Gag p24 濃度與OD450值之間呈正相關(guān),可通過繪制標(biāo)準(zhǔn)曲線求出標(biāo)本中HIV-1 Gag p24度。
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