猪圆环病毒2型Cap蛋白ELISA检测方法的建立及初步应用

将杆状病毒表达系统表达并经梯度离心纯化的PCV2Cap蛋白作为标准品，利用双抗夹心法建立了针对猪圆环病毒2型Cap蛋白的ELISA检测方法，并应用于圆环病毒2型基因工程疫苗的质控。结果表明，多抗最适包被浓度为1μg/mL，最佳封闭液为1％BSA，最佳单抗反应浓度为2μg／mL，反应时间60min，最佳二抗使用稀释度1：40000，反应时间60min。该方法与St9细胞、BSA和其他猪病病毒抗原之间无交叉反应，最低检测抗原量为5ng。

Establishment and Application of ELISA Assay for Porcine Circovirus Type 2 Cap Protein

Double-antibody-sandwich method for detecting porcine circovirus type 2(PCV2) was established using polyclonal antibody to capture antigen,monoclonal antibody to detect antigen,and PCV2 Cap protein as standard substance obtained by expressed in baculovirus system and purified with gradient centrifugation.Meanwhile this method was used for quality control of gene engineering vaccine for PCV2.Results of the optimal conditions for the assay were shown as follows:1 μg/mL of coating concentration of polyclonal antibody,1% BSA of blocking buffer,2 μg/mL of monoclonal reaction concentration with 60 min reaction time,1∶ 40000 of secondary antibody dilution with 60 min reaction time.Minimum antigen level for detection of the assay was 5 ng and it had no cross reactions with Sf9 cells,BSA,and virus antigens of other porcine diseases.