公开(公告)号：KR100430934B1

公开(公告)日：2004-05-14

申请号：KR1020010082976

申请日：2001-12-21

Applicant: 대한민국(농림축산식품부 농림축산검역본부장)

Inventor： 권창희 ,  현방훈 ,  고영준 ,  이광녕 ,  구복경 ,  엄재구 ,  나진주 ,  남향미 ,  박지용 ,  계수정 ,  김인중 ,  이세영 ,  최광석 ,  손현주 ,  주이석

IPC: C12N15/33

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和单克隆抗体中表达的重组3ABC非结构蛋白的口蹄疫诊断方法，从而比现有方法更快速和准确地诊断口蹄疫。 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由 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。通过用重组杆状病毒感染昆虫细胞表达重组3ABC非结构蛋白。 通过将在大肠杆菌中表达的重组3ABC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3F-11（KCTC 10138BP）。 从杂交瘤细胞系3F-11（KCTC 10138BP）产生单克隆抗体。 一种口蹄疫诊断方法，包括以下步骤：（1）将重组3ABC非结构蛋白稀释于包被缓冲液中，并将稀释液倒入平板中; （2）使测试血清与平板反应; （3）使与试验血清中的口蹄疫病毒抗体结合的缀合物与测试血清具有酶，放射性物质或荧光物质; 和（4）测量酶反应的强度，荧光反应或与缀合物的辐射反应。

公开(公告)号：KR1020030052858A

公开(公告)日：2003-06-27

申请号：KR1020010082976

申请日：2001-12-21

Applicant: 대한민국(농림축산식품부 농림축산검역본부장)

Inventor： 권창희 ,  현방훈 ,  고영준 ,  이광녕 ,  구복경 ,  엄재구 ,  나진주 ,  남향미 ,  박지용 ,  계수정 ,  김인중 ,  이세영 ,  최광석 ,  손현주 ,  주이석

IPC: C12N15/33

CPC classification number: C12N15/866 ,  C07K14/005 ,  C07K16/1009 ,  C07K2317/14 ,  C12N5/163 ,  C12N2770/32131 ,  G01N33/56983 ,  G01N33/577 ,  G01N2333/09

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。