公开(公告)号：KR100430935B1

公开(公告)日：2004-05-14

申请号：KR1020010082975

申请日：2001-12-21

Applicant: 대한민국(농림축산식품부 농림축산검역본부장)

Inventor： 권창희 ,  권병준 ,  고영준 ,  나진주 ,  김원일 ,  이세영 ,  주이석

IPC: C12N15/33

Abstract: PURPOSE: Diagnostic methods of foot-and-mouth disease using a recombinant 3ABC non-structural protein expressed in E. coli 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. The recombinant 3ABC non-structural protein is expressed from a recombinant E. coli transformed with a recombinant vector containing the recombinant 3ABC non-structural protein gene of SEQ ID NO: 1. 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 dilution on a plate; (2) reacting the testing serum with the plate; (3) reacting the 3ABC non-structural protein specific monoclonal antibody with the serum; (4) reacting a 3ABC non-structural protein specific monoclonal antibody binding conjugate with the monoclonal antibody; and (5) measuring intensity of the enzyme reaction, fluorescence reaction or radiation reaction with the conjugate.

Abstract translation: 目的：提供使用在大肠杆菌中表达的重组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3ABC非结构蛋白的基​​因具有SEQ ID NO：1的核苷酸序列。重组3ABC非结构蛋白由重组 用含有SEQ ID NO：1的重组3ABC非结构蛋白基因的重组载体转化大肠杆菌。通过引入在E.coli中表达的重组3ABC非结构蛋白制备杂交瘤细胞系3F-11（KCTC 10138BP） 将大肠杆菌导入动物体内，从动物体内收集免疫细胞，并将免疫细胞与癌细胞融合。 从杂交瘤细胞系3F-11（KCTC 10138BP）产生单克隆抗体。 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，并将稀释液倒入板中; （2）使测试血清与平板反应; （3）使3ABC非结构蛋白特异性单克隆抗体与血清反应; （4）使3ABC非结构蛋白特异性单克隆抗体结合缀合物与单克隆抗体反应; 和（5）测量酶反应的强度，与偶联物的荧光反应或辐射反应。

公开(公告)号：KR1020030052857A

公开(公告)日：2003-06-27

申请号：KR1020010082975

申请日：2001-12-21

Applicant: 대한민국(농림축산식품부 농림축산검역본부장)

Inventor： 권창희 ,  권병준 ,  고영준 ,  나진주 ,  김원일 ,  이세영 ,  주이석

IPC: C12N15/33

CPC classification number: C07K14/005 ,  C07K16/1009 ,  C07K2317/14 ,  C12N5/163 ,  C12N15/70 ,  C12N2770/32131 ,  G01N33/56983 ,  G01N33/577 ,  G01N2333/09

Abstract: PURPOSE: Diagnostic methods of foot-and-mouth disease using a recombinant 3ABC non-structural protein expressed in E. coli 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. The recombinant 3ABC non-structural protein is expressed from a recombinant E. coli transformed with a recombinant vector containing the recombinant 3ABC non-structural protein gene of SEQ ID NO: 1. 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 dilution on a plate; (2) reacting the testing serum with the plate; (3) reacting the 3ABC non-structural protein specific monoclonal antibody with the serum; (4) reacting a 3ABC non-structural protein specific monoclonal antibody binding conjugate with the monoclonal antibody; and (5) measuring intensity of the enzyme reaction, fluorescence reaction or radiation reaction with the conjugate.

Abstract translation: 目的：提供使用在大肠杆菌中表达的重组3ABC非结构蛋白和单克隆抗体的口蹄疫的诊断方法，从而比现有方法更快速和准确地诊断口蹄疫。 构成：编码来自韩国口蹄疫病毒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。重组体3ABC非结构蛋白由重组体 用含有SEQ ID NO：1的重组3ABC非结构蛋白基因的重组载体转化大肠杆菌。通过引入在E中表达的重组3ABC非结构蛋白来制备杂交瘤细胞系3F-11（KCTC 10138BP）。 将大肠杆菌转化成动物，从动物中收集免疫的细胞并将免疫的细胞与癌细胞融合。 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，并将稀释液倒在平板上; （2）使测试血清与板反应; （3）使3ABC非结构蛋白特异性单克隆抗体与血清反应; （4）使3ABC非结构蛋白特异性单克隆抗体结合缀合物与单克隆抗体反应; 和（5）测量酶反应的强度，荧光反应或与缀合物的辐射反应。