| 新城疫病毒HN蛋白单克隆抗体的制备及其抗原捕捉ELISA方法的建立 |
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| 引用本文： | 田莉莉,李丽.新城疫病毒HN蛋白单克隆抗体的制备及其抗原捕捉ELISA方法的建立[J].中国农学通报,2012,28(17):92-96. |
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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： | 为建立一种快速的新城疫病毒病原检测方法。本研究以原核表达的重组HN蛋白免疫7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1株能稳定分泌抗新城疫病毒HN蛋白的McAb杂交瘤细胞，命名为4E8，4E8亚类鉴定为重链属于IgG2a，轻链属于κ链。以多克隆抗体作为包被抗体、单克隆抗体4E8作为检测抗体，通过双抗夹心ELISA各个反应条件的优化，建立检测新城疫病毒抗原捕捉ELISA方法。该方法对EDSV、ITLV、IBV、IBDV不发生交叉反应，其敏感性比HA试验要高4倍以上，与RT-PCR相比较，符合率、敏感性和特异性分别为95.6%、93.3%、96.1%。本研究建立的NDV AC-ELISA有良好的重复性、敏感性和特异性，可应用于新城疫病毒感染的早期诊断。
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| 关 键 词： | 经济发展 经济发展 土地利用 保定市 |
| 收稿时间： | 2011/12/5 0:00:00 |
| 修稿时间： | 1/4/2012 12:00:00 AM |
Preparation of Monoclonal Antibodies Against HN Protein of Newcastle Disease Virus and Establishment Antigen Capture ELISA |
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| Tian Lili
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Li Li.Preparation of Monoclonal Antibodies Against HN Protein of Newcastle Disease Virus and Establishment Antigen Capture ELISA[J].Chinese Agricultural Science Bulletin,2012,28(17):92-96. |
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| Authors: | Tian Lili Li Li |
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| Institution: | Tian Lili, Li Li (College of Animal Husbandry and Veterinary Medicne, Liaoning Medical University, Jinzhou Liaoning 121000) |
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| Abstract: | To produce a rapid Newcastle disease virus （NDV） pathogen detection method. BALB/c mice were immunized with HN recombination protein and the mouse splenic cells were fused with SP2/0 cells, hybridoma cell stably secreting anti-HA McAb was screened by ELISA designated 4F8, the immunoglobulin type of McAb identification shown was IgG2a type with K chain. An antigen capture ELISA （AC-ELISA） was developed for detection of NDV using polyclonal antibody as capture antibody and specific monoclonal antibody 4F8 as detecting antibody. The AC-ELISA showed no cross-reaction with other four avian viruses （EDSV, ITLV, IBV, IBDV）, The sensitivity four times higher than HA test, Comparing with RT-PCR, the concordance, sensitivity, specificity was 95.6%, 93.3%, 96.1%, respectively. The NDV AC-ELISA had good specificity, sensitivity and repeatability, could be used for diagnosing NDV infections. |
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| Keywords: | Newcastle disease virus HN protein monoclonal antibodies antigen capture ELISA diagnosis |
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