用VP_(7)-ELISA检测牛、羊血清蓝舌病抗体的研究I:VP_(7)-ELISA检测蓝舌病抗体方法的建立

Studies on the Application of VP_(7)-ELISA for the Detection of Bluetongue Antibodies in Serum Samples from Cattle or Sheep I.Establishmeni of VP_(7)-ELISA for Bluetongue Antibodies

将表达有蓝舌病毒 ＶＰ７蛋白的 Ｓｆ２１细胞超声波处理制备 ＶＰ７抗原，建立了 ＶＰ７— ＥＬＩＳＡ检测蓝舌病抗体的方法，确定阴阳性判定临界值及最佳反应条件：待检牛血清阳性下限为３．０，羊血清阳性下限为２．４，ＶＰ７抗原包被浓度为 １： ８００，用含５％健康鸡血清的磷酸盐缓冲液作封闭液，待检血清 １：１００稀释，豚鼠抗牛羊ＩｇＧ－ＨＲＰ结合物１：５００稀释，３７℃避光显色４ ｍｉｎ～６ ｍｉｎ。试验结果表明：ＶＰ７可与２３个不同血清型ＢＴＶ抗体反应，具有较高的群特异性和敏感性。

An indirect ELISA was developed for detecting BT antibodies using VP7 antigen prepared from sonificated Sf21 cells expressing BTV-P7 protein.The cut-off was 3.0 for bovine serum and 2. 4 for ovine serum.The VP7 antigen coating concentration was 1: 800, the dilution of test serum was 1: 100; guinea pig- anti -bovine (or ovine) IgG-HRP conjugate was diluted in 1: 500. It was shown that VP7 reacted to 23 different BIV sereotypes with high group specificity and sensitivity.