| 猪细小病毒VP2蛋白单克隆抗体的制备及抗原捕捉ELISA方法的建立 |
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| 引用本文： | 田莉莉,李丽.猪细小病毒VP2蛋白单克隆抗体的制备及抗原捕捉ELISA方法的建立[J].畜牧与兽医,2012,44(7):13-17. |
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| 作者姓名： | 田莉莉 李丽 |
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| 作者单位： | 辽宁医学院畜牧兽医学院,辽宁锦州,121000 |
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| 摘 要： | 为制备猪细小病毒VP2蛋白单克隆抗体(McAb),建立检测猪细小病毒的抗原捕捉ELISA方法 (AC-ELISA),本研究以原核表达的重组VP2蛋白作为免疫原,免疫6周龄BALB/c雌鼠,取其脾细胞与骨髓瘤细胞SP2/0进行融合,经间接ELISA方法筛选,成功获得了2株能稳定分泌抗猪细小病毒VP2蛋白的McAb,命名为3C4、5F8。以多克隆抗体作为捕获抗体、单克隆抗体5F8作为检测抗体,通过双抗夹心ELISA各个反应条件的优化,建立检测猪细小病毒抗原捕捉ELISA方法。该方法与日本乙脑病毒(JEV)、猪流行性腹泻病毒(PEDV)、伪狂犬病毒(PRV)、猪繁殖与呼吸综合征病毒(PRRSV)、猪瘟病毒(CSFV)均不发生交叉反应;与RT-PCR相比较,符合率、敏感性和特异性分别为93.6%、90.9%、94.4%。本研究建立的猪细小病毒AC-ELISA有良好的重复性、敏感性和特异性,可应用于猪细小病毒感染的早期诊断。
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| 关 键 词： | 猪细小病毒 VP2蛋白 单克隆抗体 抗原捕捉ELISA |
Preparation of monoclonal antibodies against VP2 protein of porcine parvovirus and establishment of antigen capture ELISA |
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| TIAN Li-li
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LI Li.Preparation of monoclonal antibodies against VP2 protein of porcine parvovirus and establishment of antigen capture ELISA[J].Animal Husbandry & Veterinary Medicine,2012,44(7):13-17. |
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| Authors: | TIAN Li-li LI Li |
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| Institution: | TIAN Li-li,LI Li(College of Animal Husbandry and Veterinary Medicine,Liaoning Medical University,Jinzhou 121000,China) |
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| Abstract: | To produce monoclonal antibodies(McAb) against VP2 protein of porcine parvovirus(PPV) and to establish an antigen capture ELISA(AC-ELISA),BALB/c mice were immunized with VP2 recombinant protein and the mouse splenic cells were fused with SP2/0 cells.The hybridoma cell designated 3C4 and 5F8 was screened by ELISA coated with the PPV antigen.AC-ELISA was developed for detection of PPV using polyclonal antibody as capture antibody and McAb 5F8 as detecting antibody.The AC-ELISA showed no cross-reaction with other five swine viruses,including Japanese encephalitis virus(JEV),porcine epidemic diarrhea virus(PEDV),pseudorabies virus(PRV),porcine reproductive and respiratory syndrome virus(PRRSV),and classical swine fever virus(CSFV).Comparing with RT-PCR,the concordance was 93.6%,the sensitivity was 90.9%,and the specificity was 94.4%.Therefore,The PPV AC-ELISA has good specificity,sensitivity and repeatability,and could be used for diagnosis of PPV infections. |
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| Keywords: | porcine parvovirus VP2 protein monoclonal antibodies antigen capture ELISA |
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