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摘要：兔出血症是由兔出血症病毒所导致的一种兔急性、高度传染性、高度致死性的疾病。2010开始，被命名为RHDV2的变异型RHDV在我国肉兔进口国法国等国家发现并扩散，已给欧洲养兔业造成了极大地损失。目前没有有效检查方法判断其是否传播入我国，可能存在隐性感染。本课题用小鼠分别制备传统型RHDV VP60蛋白和变异型RHDV2 VP60的多克隆抗体，经间接ELISA和血凝抑制实验测定血清效价良好，经Western Blot鉴定传统型RHDV VP60制备的多克隆抗体不与新型RHDV2 VP60上特有的S1、S2片段反应，而新型RHDV2 VP60制备的多抗与这两个片段反应良好。本实验为后续建立ELISA方法以区别鉴定临床血清样品中感染的毒株，和新型疫苗的研发奠定良好的基础。28397
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毕业论文关键词：新型兔出血症病毒；VP60蛋白；多克隆抗体；鉴定
Preparation, identification and identification of polyclonal antibodies against VP60 protein of Rabbit Hemorrhagic Disease virus conventional strain and variant strain
Abstract： Rabbit Hemorrhagic Disease is an acute, highly contagious, highly lethal disease,caused by RHDV. In 2010, the variant of RHDV named RHDV2, was discovered and spread in countries such as France, which has caused considerable damage to the European rabbit industry. There is no effectiv way so far to determine whether or not it has been transmitted in our country.Recessive infection might exist. The purpose of this study is to prepare the polyclonal antibody of traditional RHDV VP60 protein and variant RHDV2 VP60 respectively. The indirect ELISA test and HI test detect the high titer and specificity.Western Blot Identificed that of polyclonal antibody prepared by traditional RHDV VP60 not reacts with the new RHDV2 VP60 special S1, S2 fragments and the new RHDV2 VP60 preparation reacts this two fragments well.This experiment establishes a good basis for the follow-up of Elisa methods to distinguish the infected strains from clinical serum samples, and the development of new vaccines.
Key words: New Rabbit hemorrhagic disease Virus; VP60 protein; polyclonal antibody;Identification
目 录
摘要 1
关键词 1
Abstract 1
Key words 1
引言 1
1 材料与方法 2
1.1 材料 2
1.1.1实验动物 2
1.1.2免疫抗原 错误！未定义书签。
1.1.3 主要试剂 2
1.1.4 设备 2
1.2 方法 3
1.2.1 兔出血症病毒传统毒株和变异毒株VP60蛋白的鉴定 3
1.2.2 实验动物免疫 4
1.2.3 血清多克隆抗体IgG纯化 4
1.2.4 多克隆抗体血凝抑制和间接ELISA效价测定 4
1.2.5 多克隆抗体Western Blot鉴定 5
2 结果 6
2.1 兔出血症病毒传统毒株和变异毒株VP60蛋白的鉴定 6
2.1.1 VP60蛋白Western Blot及血凝效价检测 7
2.1.2 结果 错误！未定义书签。
2.2 兔出血症病毒传统毒株和变异毒株VP60蛋白多克隆抗体效价测定 7
2.2.1间接ELISA测定两种多克隆抗体的效价 7
2.2.2血凝抑制测定抗体效价 7
2.2.3 结果 8
2.3 兔出血症病毒传统毒株和变异毒株VP60蛋白多克隆抗体的鉴定 8