HIV感染早期病毒p24蛋白的检测

目的:建立敏感、特异的检测血清中HIV-p24蛋白的方法,作为HIV感染早期即窗口期的监测手段。方法:用纯化的p24蛋白免疫小鼠及家兔,获得单克隆及多克隆抗体,经DEAE-52阴离子交换柱纯化后,标记辣根过氧化物酶,建立ELISA双抗体夹心及间接双抗体夹心方法,检测HIV-p24蛋白。结果:包被单抗、标记多抗或包被多抗、标记单抗,均能特异地检出系列稀释的p24蛋白,包被混合单抗较包被多抗更敏感;经标记的抗种属抗体放大可明显提高检测的敏感性。结论:建立了敏感、特异的检测p24蛋白的双抗体夹心法,间接放大方法可检出50pg/mL的HIV-p24蛋白,检测敏感性与国际同类产品相似。

Detection of p24 Antigen of HIV-1 for Early Diagnosis

Objective To establish a sensitive and specific method for detection of HIV-1 p24 antigen as early diagnosis of HIV infection. Methods Monoclonal antibody and polyclonal antibody were obtained from immunity rabbit and mice using purified HIV-1 p24 antigen. Antibodies were purified by anion-exchange column and conjugated with HRP. Double antibodies sandwich method and indirect double antibodies sandwich method to detection of p24 antigen were established. Results A sensitive and specific detecting method was selected by using with different coating antibodies conjugated antibodies and amplification systems. Conclusion Sensitive and specific double antibodies sandwich to detect p24 antigen of HIV-1 were established and 50 pg/mL p24 antigen can be detected by the indirect method.