公开(公告)号：KR1020100058222A

公开(公告)日：2010-06-03

申请号：KR1020080116959

申请日：2008-11-24

Applicant: 대한민국(농림축산식품부 농림축산검역본부장) ,  주식회사 인트론바이오테크놀로지

Inventor： 김인중 ,  현방훈 ,  윤성준 ,  이현영 ,  유진석 ,  전수연 ,  강상현 ,  표현미 ,  조수동 ,  차상호 ,  조인수

IPC: C12N15/33 ,  C12N15/10 ,  C12Q1/68

CPC classification number: Y02A50/451 ,  C12Q1/686 ,  C12Q1/04 ,  C12Q2600/16

Abstract: PURPOSE: A bovine corona virus and a method for detecting bovine virus and pestivirus are provided to enable hot-start PCR and suppress non-specific amplification. CONSTITUTION: A primer for detecting bovine diarrheal disease contains a primer for detecting bovine corona virus and a prime for detecting rotavirus. The primer for detecting bovine corona virus is a sequence of amplifying a gene encoding hemagglutinin esterase of bovine corona virus. The primer for detecting rotavirus amplifies a sequence corresponding to VP7 among genes encoding capsid gene of rotavirus. A method for detecting bovine corona virus, bovine rotavirus, and bovine diarrheal disease comprises: a step of extracting RNA, a step of performing PCR using the RNA, and a step of analyzing the PCR product.

Abstract translation: 目的：提供牛冠状病毒和检测牛病毒和瘟病毒的方法，以实现热启动PCR和抑制非特异性扩增。 构成：用于检测牛腹泻病的底漆含有检测牛冠状病毒的引物和用于检测轮状病毒的质粒。 用于检测牛冠状病毒的引物是扩增编码牛冠状病毒血凝素酯酶的基因的序列。 用于检测轮状病毒的引物在轮状病毒的编码衣壳基因的基因中扩增与VP7相对应的序列。 一种检测牛冠状病毒，牛轮状病毒和牛腹泻病的方法包括：提取RNA的步骤，使用RNA进行PCR的步骤，以及分析PCR产物的步骤。

公开(公告)号：KR100923664B1

公开(公告)日：2009-10-28

申请号：KR1020070096698

申请日：2007-09-21

Applicant: 대한민국(농림축산식품부 농림축산검역본부장) ,  주식회사 코미팜

Inventor： 현방훈 ,  표현미 ,  김인중 ,  조수동 ,  장형관 ,  문성철 ,  송재영

IPC: C12N15/64 ,  C12N15/09 ,  C07K16/00

Abstract: 본 발명은 돼지 면역글로불린 지(Immunoglobulin G; 이하 “IgG”라 한다)의 에프씨(Crystallized Fragment; 이하 “Fc”한다) 도메인의 세포 표면 발현용 벡터, 상기 벡터에 의해 형질전환된 숙주세포 및 상기 숙주세포를 이용한 돼지 질병 관련 바이러스에 대한 백신의 제조방법에 관한 것이다. 본 발명에 따른 숙주세포에 바이러스를 증식시키는 경우 바이러스 외피에 Fc가 포함됨으로써 다양한 돼지 질병 관련 바이러스에 대하여 면역능이 우수한 백신을 손쉽게 생산할 수 있도록 해준다.
돼지 IgG, Fc 유전자, 표면 발현, 벡터

公开(公告)号：KR100449908B1

公开(公告)日：2004-09-22

申请号：KR1020010082714

申请日：2001-12-21

Applicant: 대한민국(농림축산식품부 농림축산검역본부장)

Inventor： 권창희 ,  권병준 ,  고영준 ,  이세영 ,  김인중 ,  주이석

IPC: A61K39/12

Abstract: PURPOSE: A method of diagnosing vesicular stomatitis by performing an ELISA method using the recombinant envelope glycoprotein antigen of vesicular stomatitis virus(VSV) and monoclonal antibodies thereon is provided. Therefore, it permits quick, exact and specific antibody detection of the VSV and thus exact diagnosis of vesicular stomatitis through the detection. CONSTITUTION: The diagnosing method for vesicular stomatitis is achieved through a VSV detection method consisting of: attaching monoclonal antibodies against an envelope glycoprotein antigen of VSV on a solid supporter; binding the envelope glycoprotein antigen to the monoclonal antibodies attached on the solid supporter in the first step; reacting a sample containing an antibody against the envelope glycoprotein antigen with the envelope glycoprotein antigen of the second step; reacting a secondary antibody selected from the group consisting of a conjugate antibody attached with enzyme and a conjugate antibody attached with a luminescent material with the antibody of the third step.

Abstract translation: 目的：提供一种使用水疱性口炎病毒（VSV）的重组包膜糖蛋白抗原和单克隆抗体进行ELISA方法来诊断水疱性口炎的方法。 因此，它可以快速，准确和特异地检测VSV的抗体，从而通过检测来精确诊断水疱性口炎。 组成：水泡性口炎的诊断方法通过VSV检测方法实现，该方法包括：将抗VSV的包膜糖蛋白抗原的单克隆抗体附着在固体支持物上; 在第一步中将包膜糖蛋白抗原结合至附着于固体载体上的单克隆抗体; 使含有抗包膜糖蛋白抗原的抗体的样品与第二步的包膜糖蛋白抗原反应; 使选自附着有酶的缀合物抗体和附着有荧光物质的缀合物抗体的第二抗体与第三步骤的抗体反应。

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。

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。