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目的:建立快速、灵敏的检测人可溶性疱疹病毒侵入介体(HVEM)双抗体夹心ELISA。方法:在获得HVEM重组蛋白的基础上,制备兔抗人HVEM多克隆抗体,以HVEM单克隆抗体(mAb)和HVEM多克隆抗体(pAb)为双抗体,建立双抗体夹心ELISA。结果:建立的检测人可溶性HVEM双抗体夹心ELISA最低检出限为3.91μg/L,标准曲线范围7.81～250μg/L,线性方程为y=0.0021x+0.1852,R2=0.9944。回收率在89.7%～92.8%之间,平均回收率为91.4%。批内变异系数<10%,批间变异系数<15%。结论:建立的双抗体夹心ELISA快速、灵敏、简单,可满足实际工作需要。
OBJECTIVE: To establish a rapid and sensitive double antibody sandwich ELISA for the detection of human herpesvirus 2 (HSV) invasion mediator (HVEM). Methods: Based on the obtained HVEM recombinant protein, a rabbit anti-human HVEM polyclonal antibody was prepared. The double antibody sandwich ELISA was established by using the monoclonal antibody (mAb) of HVEM and the polyclonal antibody of HVEM (pAb) as the double antibody. Results: The minimum detectable limit of detection of human soluble HVEM double antibody sandwich ELISA was 3.91μg / L and the standard curve range was 7.81 ~ 250μg / L. The linear equation was y = 0.0021x + 0.1852, R2 = 0.9944. The recoveries ranged from 89.7% to 92.8% with an average recovery of 91.4%. Intra-assay CV <10%, Inter-assay CV <15%. Conclusion: The established double antibody sandwich ELISA is fast, sensitive and simple, which can meet the needs of practical work.