地方流行性牛白血病间接gp51-ELISA抗体检测试剂盒的研制

Development of indirect gp51-enzyme-linked immunosorbent assay kit for detecting antibody against bovine leukemia virus

利用重组牛白血病痛毒gp51蛋白作为包被抗原研制了gp51-ELISA诊断试剂盒。试剂盒主要成分包括抗原gp51包被96孔板、HRP标记的兔抗牛IgG和牛白血病阴性、阳性标准血清。对试剂盒进行了特异性、灵敏度、重复性及保存期试验。结果表明，试剂盒特异性好，灵敏度是AGIDT的4-8倍，批间和批内的变异系数分别小于10％和15％，于-20℃保存至少能保持6个月检测结果稳定。与法国Synbiotics公司同类试剂盒进行比对试验，诊断敏感性、特异性和符合率分别为75％、96.1％和94.6％。对采集于山东济南的164份奶牛血清和进口澳大利亚奶牛的398份血清进行检测，抗体阳性率分别6.10％和4.77％。

An enzyme-linked immunosobent assay test kit was developed using purified BLV gpS1 protein as coating antigen for detecting antibody against bovine leukemia virus. The main components of the kit are one 96-well plate coated by gp51 protein, HRP labelled rabbit anti-bovine IgG, the standard positive/negative serum. The specificity,sensitivity,reproducibility and keeping time of the kit were tested. The sensitivity of the kit was 4-8 fold of the AGIDT;the coefficiens of variance among and between batch were less than 10% and 15% respectively;stablity for at least six months stored at -20℃. The kit was compared side by side with BLV ELISA kit produced by France Synbioties Company, the results showed that the sensitivity, specificity and coincidence were 75%, 96.1% and 94.6%, respectively. The kit was used to test 164 sera from cows in Jinan city, Shandong province and 398 sera from Australia import cows, 6.10% and 4.77% of them were positive, respectively.