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首页> 外国专利> SWINE FEVER VIRUS ERNS PROTEIN AND METHOD FOR SCREENING AND DETECTING A SWINE FEVER VIRUS ANTIBODY THAT SPCIFICALLY REACTS WITH THE SAME

SWINE FEVER VIRUS ERNS PROTEIN AND METHOD FOR SCREENING AND DETECTING A SWINE FEVER VIRUS ANTIBODY THAT SPCIFICALLY REACTS WITH THE SAME

机译：猪瘟病毒蛋白及其筛选和检测与猪瘟病毒相同反应的猪瘟病毒抗体的方法

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A method for screening and detecting a swine fever virus antibody is provided to easily screen and diagnose infected swine fever virus antibody by using a monoclonal antibody specifically reacting with a recombinant swine fever virus Erns protein and secure constant sensitivity regardless of all genotypes of the swine fever virus when using all three kinds of recombinant protein. A recombinant swine fever virus Erns is prepared by expressing a gene encoding three Erns proteins among swine fever viruses having various genotypes using baculovirus. A method for preparing a monoclonal antibody specifically reacting to the Erns comprises the steps of: (a) constructing a swine fever virus LOM strain-derived Erns recombinant gene using a baculovirus expression system; and (b) producing the monoclonal antibody using the recombinant Erns protein. A method for screening and detecting a swine fever virus antibody comprises the steps of: (a) mixing three kinds of Erns recombinant proteins at proper concentration to prepare an antigen cocktail; (b) after diluting the antigen cocktail into a coating buffer solution, pouring it in a plate to be absorbed into the plate; (c) washing the recombinant antigen not-absorbed into the plate to remove it; (d) adding a sample to be tested to the plat to allow the reaction; (e) washing the sample to be tested not specifically bound to the recombinant Erns antigen protein absorbed into the plate to remove it; (f) adding a monoclonal antibody not being bound to the enzyme but being specifically reacted to the Erns protein to allow the reaction; (g) washing the monoclonal antibody not being bound to the recombinant Erns antigen protein absorbed into the plate to remove it; and (h) adding a substrate reacting with the enzyme to determine the swine fever virus infection.
机译：提供一种用于筛选和检测猪瘟病毒抗体的方法，以通过使用与重组猪瘟病毒Erns蛋白特异性反应的单克隆抗体轻松地筛选和诊断受感染的猪瘟病毒抗体，并且不管猪瘟的所有基因型如何，都可确保恒定的敏感性病毒时使用所有三种重组蛋白。通过使用杆状病毒在具有各种基因型的猪瘟病毒中表达编码三种Erns蛋白的基因来制备重组猪瘟病毒Erns。一种制备与Erns特异性反应的单克隆抗体的方法包括以下步骤：（a）使用杆状病毒表达系统构建猪瘟病毒LOM株衍生的Erns重组基因。 （b）使用重组Erns蛋白生产单克隆抗体。一种猪瘟病毒抗体的筛选和检测方法，包括以下步骤：（a）以适当的浓度混合三种Erns重组蛋白以制备抗原混合物。 （b）将抗原混合物稀释到包被缓冲溶液中后，倒入平板中以吸收到平板中; （c）洗涤未吸收到板中的重组抗原以除去它; （d）将待测试的样品添加到平板上以进行反应; （e）洗涤未特异性结合吸收到板中的重组Erns抗原蛋白的待测样品以除去它; （f）添加不与酶结合但与Erns蛋白特异性反应的单克隆抗体以允许反应; （g）洗涤不与吸收到平板中的重组Erns抗原蛋白结合的单克隆抗体以除去它; （h）添加与酶反应的底物以确定猪瘟病毒感染。

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