猪圆环病毒2型ELISA抗体检测试剂盒的研制及初步应用

Development and Primary Application of ELISA KIT for the Detection of Antibody against Porcine Circovirus Type 2

关键词:  猪圆环病毒2型,ELISA,捕获包被,抗体检测

Key words:  Porcine  circovirus type 2,ELISA,Capture  coating,Antibody  detection

摘要:

用PCV2 Cap蛋白单克隆抗体预包被酶标板，将杆状病毒表达系统表达的PCV2 Cap蛋白作为检测用抗原，利用捕获包被法建立了间接ELISA法用于猪圆环病毒2型抗体的检测。通过优化ELISA条件，研制试剂盒并应用于圆环病毒2型疫苗的免疫效果检验。利用研制的试剂盒与IFA、商品化进口和国产同类试剂盒对178份猪血清进行平行检测，总符合率分别为96.63%、97.75%和84.27%，与其他几种常见猪病毒抗体阳性血清无交叉反应。试剂盒批内、批间变异系数分别为2.35%~4.06%和4.87%~6.54%，2~8 ℃保存15个月稳定。基于用单抗预包被后再包被检测抗原，本试剂盒检测样品抗体的灵敏度和稳定性得以显著提高，因此适合于对不同来源猪圆环病毒2型疫苗人工免疫猪血清抗体进行检测，从而对猪圆环病毒2型疫苗的免疫效果进行评价。

Abstract:

With capture coating method, indirect ELISA method for detecting porcine circovirus type 2 (PCV2) antibody was established using PCV2 cap protein monoclonal antibody to pre-coat ELISA plate and using PCV2 cap protein expressed in baculovirus system as detecting antigen. The ELISA was optimized and the kit was developed for checking the immune results of PCV2 vaccines. Parallel testing on 178 porcine serums between the kit and IFA, imported and domestic commercial kits of the same type showed the total accordance rates being 96.63%, 97.75% and 84.27%, respectively, and there was no cross reactions between the kit and the positive serums of other several common porcine virus antibodies. The coefficients of variation of intra-batch and inter-batch kits were 2.35%~4.06% and 4.87%~6.54%, respectively. The shelve life of the kit was 15 months when storing at 2~8 ℃. Based on pre-coating monoclonal antibody and followed by coating detecting antigen, the sensitivity and stability of the kit were significantly improved, hence suitable for detecting the antibody in the serums of porcines immunized with PCV2 vaccines from different sources and consequently to evaluate the immune effects of PCV2 vaccines.