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    • 6. 发明公开
    • 돼지콜레라 바이러스 Erns 단백질 및 이에 특이적으로반응하는 단클론항체를 이용한 돼지콜레라 항체 감별진단방법
    • SWINE FEVER VIRUS ERNS蛋白质和筛选和检测与其相似的SWINE FEVER病毒抗体的方法
    • KR1020070116382A
    • 2007-12-10
    • KR1020060050357
    • 2006-06-05
    • 대한민국(농림축산식품부 농림축산검역본부장)주식회사 메디안디노스틱
    • 송재영김병한탁동섭최은진임성인한규하주후돈장병식이현정박경민
    • C12N15/33C12N15/09C12N15/10
    • 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.
    • 提供了一种筛选和检测猪瘟病毒抗体的方法,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筛选和诊断感染的猪瘟病毒抗体,无论猪瘟的所有基因型如何 使用全部三种重组蛋白的病毒。 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蛋白质的基因来制备重组猪瘟病毒Erns。 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菌株衍生的Erns重组基因; 和(b)使用重组Erns蛋白产生单克隆抗体。 筛选和检测猪瘟病毒抗体的方法包括以下步骤:(a)以适当浓度混合三种Erns重组蛋白以制备抗原混合物; (b)将抗原混合物稀释到涂布缓冲溶液中后,将其倒入板中吸收到板中; (c)将未吸收的重组抗原洗涤到板中以除去它; (d)将待测试样品加入平板以允许反应; (e)洗涤待测试的样品,未特异性地结合到吸收到板中的重组Erns抗原蛋白质以除去它; (f)加入未结合酶的单克隆抗体,但与Erns蛋白特异性反应以进行反应; (g)洗涤未结合到吸附到板中的重组Erns抗原蛋白的单克隆抗体以除去它; 和(h)加入与酶反应的底物以确定猪瘟病毒感染。