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    구제역 바이러스 7가지 혈청형 진단용 프라이머 세트 및 이의 용도
    1.
    发明申请
    구제역 바이러스 7가지 혈청형 진단용 프라이머 세트 및 이의 용도 审中-公开

    公开(公告)号：WO2019151764A1

    公开(公告)日：2019-08-08

    申请号：PCT/KR2019/001274

    申请日：2019-01-30

    Applicant: 경북대학교 산학협력단 ,  대한민국(농림축산식품부 농림축산검역본부장)

    Inventor： 박최규 ,  임다래 ,  위성환 ,  구복경 ,  나진주 ,  유소윤 ,  전효성

    IPC: C12Q1/70

    Abstract: 본 발명은 구제역 바이러스 7가지 혈청형 진단용 프라이머 세트 및 이의 용도에 관한 것이다. 본 발명의 구제역 바이러스 검출용 프라이머 세트는, 낮은 농도의 시료만으로도 7가지 구제역바이러스의 혈청형을 검출할 수 있어 민감도가 높고, 특이도가 우수하다. 또한, 본 발명의 구제역 바이러스 검출용 프라이머 세트를 이용한 RT-LAMP법을 이용하면 기존의 진단법과는 달리 등온조건에서 유전자 증폭이 이루어지기 때문에 검사시간을 단축할 수 있으며, 저가의 장비로도 반응이 가능하다. 따라서, 고가의 장비, 시약 및 전문인력이 확보되지 않은 진단실이나 현장에서도 구제역 바이러스를 신속하고 효과적으로 진단할 수 있는 바, 국내외 바이러스 진단 시장에서 용이하게 활용될 수 있다.

    재조합 구제역 2Ｃ 비구조단백질 항원 및 단클론항체를이용한 구제역 진단방법
    7.
    发明授权
    재조합 구제역 2Ｃ 비구조단백질 항원 및 단클론항체를이용한 구제역 진단방법 有权
    재조합구제역2C비구조단백질항원및단클론항체를이용한구제역진단방재

    公开(公告)号：KR100449906B1

    公开(公告)日：2004-09-22

    申请号：KR1020010082977

    申请日：2001-12-21

    Applicant: 대한민국(농림축산식품부 농림축산검역본부장)

    Inventor： 현방훈 ,  엄재구 ,  구복경 ,  이광녕 ,  김인중 ,  고영준 ,  권창희 ,  주이석 ,  안수환

    IPC: C12N15/33

    Abstract: PURPOSE: A diagnostic method of foot-and-mouth disease using a recombinant FMCV 2C non-structural protein and a monoclonal antibody is provided, thereby more rapidly and accurately diagnosing the foot-and-mouth disease than the prior methods. CONSTITUTION: A gene encoding a recombinant FMCV 2C non-structural protein derived from Korean foot-and-mouth disease virus O/SKR/2000 has the nucleotide sequence of SEQ ID NO: 1. A recombinant vector is prepared by cloning the gene encoding the recombinant FMCV 2C non-structural protein of SEQ ID NO: 1. The recombinant FMCV 2C non-structural protein is expressed by transformation of a cell with the recombinant vector. A hybridoma cell line(KCTC 10137BP) is prepared by introducing the recombinant FMCV 2C non-structural protein into an animal, collecting an immunized cell from the animal and fusing the immunized cell with a cancer cell. A recombinant FMCV 2C non-structural protein specific monoclonal antibody is produced from the hybridoma cell line(KCTC 10137BP). A diagnostic method of foot-and-mouth disease comprises the steps of: (1) diluting the recombinant FMCV 2C non-structural protein specific monoclonal antibody in a coating buffer solution and pouring the diluate on a plate; (2) washing the plate to remove unattached monoclonal antibodies; (3) reacting the recombinant FMCV 2C non-structural protein with the plate; (4) washing the plate to remove unreacted recombinant FMCV 2C non-structural proteins; (5) reacting the testing serum with the plate; (6) washing the plate to remove unreacted testing serum; (7) reacting a conjugate, which binds with an antibody for foot-and-mouth disease virus in the testing serum and has an enzyme, a radioactive material or a fluorescent material, with the testing serum; and (8) measuring intensity of the enzyme reaction, fluorescence reaction or radiation reaction with the conjugate.

    Abstract translation: 目的：提供使用重组FMCV 2C非结构蛋白和单克隆抗体的口蹄疫诊断方法，从而比现有方法更快速和更准确地诊断口蹄疫。 构成：编码来源于韩国口蹄疫病毒O / SKR / 2000的重组FMCV 2C非结构蛋白的基​​因具有SEQ ID NO：1的核苷酸序列。重组载体通过克隆编码 重组FMCV 2C非结构蛋白SEQ ID NO：1。通过用重组载体转化细胞表达重组FMCV 2C非结构蛋白。 通过将重组FMCV 2C非结构蛋白导入动物中，从动物中收集免疫的细胞并将免疫的细胞与癌细胞融合来制备杂交瘤细胞系（KCTC 10137BP）。 从杂交瘤细胞系（KCTC 10137BP）产生重组FMCV 2C非结构蛋白特异性单克隆抗体。 一种口蹄疫诊断方法，包括以下步骤：（1）将重组FMCV 2C非结构蛋白特异性单克隆抗体稀释于包被缓冲液中，并将稀释液倒入平板中; （2）清洗平板以去除未附着的单克隆抗体; （3）使重组FMCV 2C非结构蛋白与平板反应; （4）洗涤平板以除去未反应的重组FMCV 2C非结构蛋白; （5）使测试血清与平板反应; （6）洗涤平板以除去未反应的测试血清; （7）使与试验血清中的口蹄疫病毒抗体结合的缀合物与试验血清具有酶，放射性物质或荧光物质的步骤; 和（8）测量酶反应的强度，与缀合物的荧光反应或辐射反应。

    재조합 구제역 2Ｃ 비구조단백질 항원 및 단클론항체를이용한 구제역 진단방법
    8.
    发明公开
    재조합 구제역 2Ｃ 비구조단백질 항원 및 단클론항체를이용한 구제역 진단방법 有权
    使用重组FMCV 2C非结构蛋白和单克隆抗体的足球疾病诊断方法

    公开(公告)号：KR1020030052859A

    公开(公告)日：2003-06-27

    申请号：KR1020010082977

    申请日：2001-12-21

    Applicant: 대한민국(농림축산식품부 농림축산검역본부장)

    Inventor： 현방훈 ,  엄재구 ,  구복경 ,  이광녕 ,  김인중 ,  고영준 ,  권창희 ,  주이석 ,  안수환

    IPC: C12N15/33

    CPC classification number: C12N5/163 ,  C07K16/1009 ,  C07K2317/14 ,  C12N2770/32131 ,  G01N33/56983 ,  G01N33/577 ,  G01N2333/09

    Abstract: PURPOSE: A diagnostic method of foot-and-mouth disease using a recombinant FMCV 2C non-structural protein and a monoclonal antibody is provided, thereby more rapidly and accurately diagnosing the foot-and-mouth disease than the prior methods. CONSTITUTION: A gene encoding a recombinant FMCV 2C non-structural protein derived from Korean foot-and-mouth disease virus O/SKR/2000 has the nucleotide sequence of SEQ ID NO: 1. A recombinant vector is prepared by cloning the gene encoding the recombinant FMCV 2C non-structural protein of SEQ ID NO: 1. The recombinant FMCV 2C non-structural protein is expressed by transformation of a cell with the recombinant vector. A hybridoma cell line(KCTC 10137BP) is prepared by introducing the recombinant FMCV 2C non-structural protein into an animal, collecting an immunized cell from the animal and fusing the immunized cell with a cancer cell. A recombinant FMCV 2C non-structural protein specific monoclonal antibody is produced from the hybridoma cell line(KCTC 10137BP). A diagnostic method of foot-and-mouth disease comprises the steps of: (1) diluting the recombinant FMCV 2C non-structural protein specific monoclonal antibody in a coating buffer solution and pouring the diluate on a plate; (2) washing the plate to remove unattached monoclonal antibodies; (3) reacting the recombinant FMCV 2C non-structural protein with the plate; (4) washing the plate to remove unreacted recombinant FMCV 2C non-structural proteins; (5) reacting the testing serum with the plate; (6) washing the plate to remove unreacted testing serum; (7) reacting a conjugate, which binds with an antibody for foot-and-mouth disease virus in the testing serum and has an enzyme, a radioactive material or a fluorescent material, with the testing serum; and (8) measuring intensity of the enzyme reaction, fluorescence reaction or radiation reaction with the conjugate.

    Abstract translation: 目的：提供使用重组FMCV 2C非结构蛋白和单克隆抗体的口蹄疫诊断方法，从而比现有方法更快速，准确地诊断口蹄疫。 构成：编码来自韩国口蹄疫病毒O / SKR / 2000的重组FMCV 2C非结构蛋白的基​​因具有SEQ ID NO：1的核苷酸序列。通过克隆编码 SEQ ID NO：1的重组FMCV 2C非结构蛋白。通过用重组载体转化细胞来表达重组FMCV 2C非结构蛋白。 通过将重组FMCV 2C非结构蛋白引入动物中，从动物中收集免疫的细胞并将免疫的细胞与癌细胞融合，来制备杂交瘤细胞系（KCTC 10137BP）。 从杂交瘤细胞系（KCTC 10137BP）产生重组FMCV 2C非结构蛋白特异性单克隆抗体。 口蹄疫的诊断方法包括以下步骤：（1）在包衣缓冲溶液中稀释重组FMCV 2C非结构蛋白特异性单克隆抗体，将稀释物倒入板上; （2）洗涤板以除去未附着的单克隆抗体; （3）使重组FMCV 2C非结构蛋白与板反应; （4）洗涤板以除去未反应的重组FMCV 2C非结构蛋白; （5）使测试血清与板反应; （6）洗板以除去未反应的检测血清; （7）使与测试血清中的口蹄疫病毒抗体结合并具有酶，放射性物质或荧光材料的缀合物与测试血清反应; 和（8）测量酶反应的强度，荧光反应或与缀合物的辐射反应。

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