| 摘要: |
| 将杆状病毒表达系统表达并经梯度离心纯化的PCV2 Cap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1 μg/mL，最佳封闭液为1% BSA，最佳单抗反应浓度为2 μg/mL，反应时间60 min，最佳二抗使用稀释度1∶40000，反应时间60 min。该方法与Sf9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5 ng。 |
| 关键词: 猪圆环病毒2型 衣壳蛋白Cap 检测 夹心法 |
| DOI： |
| 投稿时间：2012-02-03修订日期：2012-02-28 |
| 基金项目: |
|
| Establishment and Application of ELISA Assay for Porcine Circovirus Type 2 Cap Protein |
| liaoyuanyuan |
| (wuhanzhongboshengwugufenyouxiangongsi) |
| Abstract: |
| 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 that was obtained by expressed in baculovirus system and purified with gradient centrifugation. The method is to be used for quality control of gene engineering vaccine for PCV2. Results showed the optimal conditions for the assay were: coating concentration of polyclonal antibody 1μg/ml,blocking buffer 1%BSA,monoclonal reaction concentration 2μg/ml with 60min reaction time,secondary antibody dilution 1:40,000 with 60min reaction time,and antigen reaction time 60min. The assay has a minimum antigen for detection of 5ng and has no cross reactions with Sf9 cells, BSA, and virus antigens of other porcine diseases. |
| Key words: Porcine circovirus type 2 Capsid protein detection Sandwich |
