| 猪圆环病毒3型Cap蛋白单克隆抗体的制备及其在免疫组化试验中的初步应用 |
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| 引用本文： | 赵冬,王小波,高清清,郇长超,王万彬,高崧,刘秀梵. 猪圆环病毒3型Cap蛋白单克隆抗体的制备及其在免疫组化试验中的初步应用[J]. 中国预防兽医学报, 2019, 0(2): 174-178 |
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| 作者姓名： | 赵冬 王小波 高清清 郇长超 王万彬 高崧 刘秀梵 |
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| 作者单位： | 农业部禽用生物制品创制重点实验室;农业部畜禽传染病学重点开放实验室;江苏省动物重要疫病与人兽共患病防控协同创新中心;扬州大学农业科技发展研究院;扬州大学兽医学院;江苏省研究生工作站 |
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| 基金项目： | 江苏现代农业(生猪)产业技术体系疾病防控创新团队(SXGC[2017]231);国家重点研发计划(2016YFD0500704-2);江苏高校优势学科建设工程(PAPD) |
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| 摘 要： | 为制备猪圆环病毒3型(PCV3)Cap蛋白单克隆抗体(MAb),并初步应用于感染细胞或组织样品中PCV3抗原的间接免疫荧光试验(IFA)或免疫组化(IHC)检测,本研究将PCV3 Cap蛋白的去核定位信号肽基因克隆于原核表达载体pET-28a中,经IPTG诱导表达了具有免疫原性的融合蛋白。以表达的重组蛋白免疫BALB/c小鼠,共获得8株能够稳定分泌针对PCV3 Cap蛋白的MAb杂交瘤细胞株。Western blot与IFA试验结果显示,8株MAbs均能够与真核表达的重组Cap蛋白反应。选取3株杂交瘤细胞制备3株MAbs,小鼠腹水效价分别为1∶409 600、1∶204 800和1∶409 600。利用3株MAbs对自然感染PCV3临床病猪的腹股沟淋巴结进行IHC检测,结果显示3株MAbs的腹水均能够与感染PCV3的临床样品发生特异性的免疫反应。本研究制备的MAbs具有较好的特异性,为深入研究Cap蛋白的结构和PCV3快速诊断奠定了基础。
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| 关 键 词： | 猪圆环病毒3型 CAP蛋白 单克隆抗体 免疫组化 |
Development of monoclonal antibodies against Cap protein of porcine circovirus type 3 and its preliminary application in immunohistochemical assay |
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| ZHAO Dong,WANG Xiao-bo,GAO Qing-qing,HUAN Chang-chao,WANG Wan-bin,GAO Song,LIU Xiu-fan. Development of monoclonal antibodies against Cap protein of porcine circovirus type 3 and its preliminary application in immunohistochemical assay[J]. Chinese Journal of Preventive Veterinary Medicine, 2019, 0(2): 174-178 |
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| Authors: | ZHAO Dong WANG Xiao-bo GAO Qing-qing HUAN Chang-chao WANG Wan-bin GAO Song LIU Xiu-fan |
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| Affiliation: | (Key Laboratory of Avian Bioproducts Development, Ministry of Agriculture, Yangzhou 225009, China;Key Laboratory of Animal Infectious Diseases, Yangzhou 225009, China;Jiangsu Co-innovation Center for Prevention and Control of Important Animal Infectious Diseases and Zoonoses, Yangzhou 225009, China;Institutes of Agricultural Science and Technology Development, Yangzhou 225009, China;College of Veterinary Medicine, Yangzhou University, Yangzhou 225009, China;Postgraduates Training Station of Jiangsu Province, Taizhou 225511, China) |
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| Abstract: | ZHAO Dong;WANG Xiao-bo;GAO Qing-qing;HUAN Chang-chao;WANG Wan-bin;GAO Song;LIU Xiu-fan(Key Laboratory of Avian Bioproducts Development, Ministry of Agriculture, Yangzhou 225009, China;Key Laboratory of Animal Infectious Diseases, Yangzhou 225009, China;Jiangsu Co-innovation Center for Prevention and Control of Important Animal Infectious Diseases and Zoonoses, Yangzhou 225009, China;Institutes of Agricultural Science and Technology Development, Yangzhou 225009, China;College of Veterinary Medicine, Yangzhou University, Yangzhou 225009, China;Postgraduates Training Station of Jiangsu Province, Taizhou 225511, China) |
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| Keywords: | porcine circovirus type 3 capsid protein monoclonal antibody immunohistochemistry |
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