犬狂犬病病毒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 Canine Rabies Virus

关键词:  狂犬病病毒  ELISA  试剂盒  抗体检测

Key words:  rabies virus  ELISA  kit  antibody detection

摘要:

用狂犬病毒单克隆抗体预包被酶标板，将纯化的狂犬病毒作为检测用抗原，利用捕获包被法建立了间接ELISA法用于犬狂犬病病毒抗体的检测。通过优化ELISA条件，研制试剂盒并应用于犬狂犬疫苗的免疫效果检验。利用研制的试剂盒与RFFIT、商品化同类试剂盒对30份犬血清进行平行检测，总符合率分别为90%和93.33%，与其它几种常见犬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1.45%～5.79%和2.35%～7.21%，2~8 ℃保存12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，因此适合于不同来源狂犬疫苗人工免疫犬血清的RV抗体水平监测。

Abstract:

With capture coating method, indirect ELISA method for detecting rabies virus(RV) antibody was established using RV monoclonal antibody to pre-coat ELISA plate and using RV as detecting antigen. The ELISA was optimized and the kit was developed for checking the immune results of RV vaccines. Parallel testing on 30 canine serums among the kit, RFFIT and commercial kits of the same type showed the total accordance rates being 90% and 93.33%, respectively, and there was no cross reactions between the kit and the positive serums of other several common caninevirus antibodies. The coefficients of variation of intra-batch and inter-batch kits were 1.45%～5.79% and 2.35%～7.21%, respectively. The shelve life of the kit was 12 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 canines immunized with RV vaccines from different sources.