구제역 바이러스 7가지 혈청형 진단용 프라이머 세트 및 이의 용도

    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

    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
    4.
    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

    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
    5.
    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

    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。

    곤충세포 발현 재조합 3ABC 비구조단백질 항원 및 단클론항체를 이용한 구제역 진단방법
    9.
    发明公开
    곤충세포 발현 재조합 3ABC 비구조단백질 항원 및 단클론항체를 이용한 구제역 진단방법 有权
    使用在INSEC细胞和单克隆抗体中表达的重组蛋白3ABC非结构蛋白的足球疾病的诊断方法

    公开(公告)号:KR1020030052858A

    公开(公告)日:2003-06-27

    申请号:KR1020010082976

    申请日:2001-12-21

    Abstract: PURPOSE: Diagnostic methods of foot-and-mouth disease using recombinant 3ABC non-structural protein expressed in insect cells 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 3ABC 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 baculovirus to be expressed in insect cells is prepared by co-transfection with a recombinant vector containing the recombinant 3ABC non-structural protein gene of SEQ ID NO: 1. The recombinant 3ABC non-structural protein is expressed by infection of the insect cells with the recombinant baculovirus. A hybridoma cell line 3F-11(KCTC 10138BP) is prepared by introducing the recombinant 3ABC non-structural protein expressed in E. coli into an animal, collecting an immunized cell from the animal and fusing the immunized cell with a cancer cell. A monoclonal antibody is produced from the hybridoma cell line 3F-11(KCTC 10138BP). A diagnostic method of foot-and-mouth disease comprises the steps of: (1) diluting the recombinant 3ABC non-structural protein in a coating buffer solution and pouring the diluate on a plate; (2) reacting the testing serum with the plate; (3) 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 (4) measuring intensity of the enzyme reaction, fluorescence reaction or radiation reaction with the conjugate.

    Abstract translation: 目的:提供使用在昆虫细胞中表达的重组3ABC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的重组体3ABC非结构蛋白的基​​因具有SEQ ID NO:1的核苷酸序列。在昆虫细胞中表达的重组杆状病毒通过 用含有SEQ ID NO:1的重组3ABC非结构蛋白基因的重组载体共转染。重组体3ABC非结构蛋白用重组杆状病毒感染昆虫细胞表达。 通过将在大肠杆菌中表达的重组3ABC非结构蛋白质导入动物中,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3F-11(KCTC 10138BP)。 从杂交瘤细胞系3F-11(KCTC 10138BP)产生单克隆抗体。 口蹄疫的诊断方法包括以下步骤:(1)在包被缓冲溶液中稀释重组体3ABC非结构蛋白,将稀释液倒入平板上; (2)使测试血清与板反应; (3)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(4)测量酶反应的强度,荧光反应或与缀合物的辐射反应。

    부루셀라 케니스 변이균주(M-)를 이용한 개 부루셀라병의 신속평판응집반응 진단킷트
    10.
    发明授权
    부루셀라 케니스 변이균주(M-)를 이용한 개 부루셀라병의 신속평판응집반응 진단킷트 失效
    使用BRUCELLA CANIS突变菌株无效的犬类布鲁氏病快速病害解决方案

    公开(公告)号:KR100207937B1

    公开(公告)日:1999-07-15

    申请号:KR1019960034501

    申请日:1996-08-20

    Abstract: 본 발명은 개 부루셀라병의 진단에 있어서, 기존의 개발된 혈청학적 진단법의 단점인 부루셀라(Brucella) 균의 점조성(mucoid)으로 인한 자가응집반응 및 교차반응을 줄이기 위하여 부루셀라 케니스 변이균주(Brucella canis mutant strain Mucoidless)를 이용한 개 부루셀라병의 신속평판응집반응 진단킷트에 관한 것이다.
    부루셀라 케니스 변이균주는 부루셀라 케니스 균주(Brucella canis strain)에 비하여 병원성 감소, 비특이적인 응집성 감소, 높은 소수성 등에서 큰 차이를 나타내고, PH 4~6의 범위에서 빠른 시간에 응집하는 결과를 나타내어 현재 피트만모르사에서 사용하고 있는 부루셀라 오비스(B. ovis) 균주보다고 훨씬 신속하게 반응하는 결과를 나타내어 응집반응 원리를 이용한 혈청학적 진단시 비특이반응을 줄이는데 매우 효율적이다.
    부루셀라 케니스 변이균주를 이용한 신속평판반응집반응법은 진단효율도 높을뿐 아니라 1시간이내에 검출할 수 있을 정도로 신속하고, 사용방법도 간편함을 특징으로 한다.

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