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：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：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。

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。

公开(公告)号：KR101602921B1

公开(公告)日：2016-03-11

申请号：KR1020130110387

申请日：2013-09-13

Applicant: 대한민국(농림축산식품부 농림축산검역본부장)

Inventor： 박종현 ,  이광녕 ,  김수미 ,  김래형 ,  이여주 ,  박정남 ,  이서용 ,  고영준 ,  이향심 ,  서민구 ,  탁동섭 ,  김병한

IPC: C12N15/41 ,  C12N15/63 ,  C12N7/01 ,  A61K39/125

Abstract: 본발명은구제역 SAT 3형 SEZ 지역형의방어항원단백질을코딩하는유전자가삽입된재조합플라스미드, 상기재조합플라스미드를이용하여제조한재조합구제역바이러스및 상기재조합바이러스를유효성분으로함유하는구제역예방용백신조성물에관한것이다. 본발명의재조합구제역바이러스는구제역 SAT 3형 SEZ 지역형바이러스의방어항원인 P1 단백질을발현하는효과가우수하므로, 구제역 SAT 3형(SAT3/2sa57/59 [SAT3/SA/57/59])을비롯한구제역의백신용으로유용하게사용될수 있다.

公开(公告)号：KR101578425B1

公开(公告)日：2015-12-18

申请号：KR1020130088492

申请日：2013-07-26

Applicant: 대한민국(농림축산식품부 농림축산검역본부장)

Inventor： 박종현 ,  이광녕 ,  김수미 ,  김래형 ,  이여주 ,  박정남 ,  이서용 ,  고영준 ,  이향심 ,  서민구 ,  탁동섭 ,  김병한

IPC: C12N7/01 ,  C12N15/41 ,  A61K39/125

Abstract: 본발명은구제역 O형백신주 Manisa의전체유전자부위중에서, 상기구제역 O형백신주 Manisa의방어항원인 P1 단백질을코딩하는유전자가, 구제역 SAT1형 WZ 지역형바이러스의방어항원인 P1 단백질을코딩하는유전자로치환된재조합유전자가삽입된플라스미드를포함하는재조합구제역바이러스및 그의제조방법을제공한다.

公开(公告)号：KR101566375B1

公开(公告)日：2015-11-09

申请号：KR1020130120382

申请日：2013-10-10

Applicant: 대한민국(농림축산식품부 농림축산검역본부장)

Inventor： 이광녕 ,  홍장관 ,  김수미 ,  박종현 ,  고영준 ,  이향심 ,  조인수

IPC: C12N15/11 ,  C12Q1/68

Abstract: 본발명은구제역바이러스검출용핵산칩과그 탐침의서열에관한것으로, 구체적으로혈청형별각 구제역바이러스에특이적인 16종의탐침이포함되어혈청형별구제역바이러스유래핵산과특이적으로교잡함으로써시료내의구제역바이러스의유무및 구제역바이러스의혈청형을구분할수 있는구제역바이러스검출용인공핵산칩에관한것이다. 본발명에따르면, 시료로부터구제역바이러스를매우손쉽고빠르게타입별로검출할수 있으며, 타입이다른여러종의구제역바이러스가동시에존재하더라도정확하게검출할수 있다. 이는기존염기서열분석방법에비해소요되는시간과인력, 비용을상당히줄일수 있는방법이며, 여러종류의바이러스도동시에검출할수 있으므로매우개선된방법이라할 수있다. 또한, 새롭게변이된바이러스가나타날경우, 이에대한탐침을별도로제착하여본 발명의핵산칩에추가하는방법으로손쉽게업데이트할수 있다는장점도있다. 본발명의구제역바이러스검출용핵산칩, 이핵산칩을사용하여구제역바이러스를검출하는방법및 구제역바이러스검출용키트는특히우리나라나주변국가에서발생하는구제역에빠르게대처하는데큰 도움이될 것이라고판단된다.

公开(公告)号：KR101566371B1

公开(公告)日：2015-11-09

申请号：KR1020130110325

申请日：2013-09-13

Applicant: 대한민국(농림축산식품부 농림축산검역본부장)

Inventor： 박종현 ,  이광녕 ,  김수미 ,  김래형 ,  이여주 ,  박정남 ,  고영준 ,  이향심 ,  서민구 ,  탁동섭 ,  김병한

IPC: C12N15/41 ,  C12N15/63 ,  C12N7/01

Abstract: 본발명은구제역아시아1 혈청형의표준백신바이러스의방어항원을발현하는재조합구제역바이러스및 이의제조방법에관한것이다. 본발명에따른재조합구제역바이러스는 As1/Shamir/89의 VP1 단백질또는 VP1 단백질의에피토프를발현하는효과가우수하여, 구제역아시아1형을포함하는구제역의예방에유용하게사용될수 있다.

公开(公告)号：KR1020150047834A

公开(公告)日：2015-05-06

申请号：KR1020130127801

申请日：2013-10-25

Applicant: 대한민국(농림축산식품부 농림축산검역본부장)

Inventor： 박종현 ,  김수미 ,  이광녕 ,  초가기 ,  김래형 ,  이여주 ,  박정남 ,  신연경 ,  고영준 ,  이향심 ,  서민구 ,  탁동섭 ,  김병한

IPC: C12N7/01 ,  A61K39/135

CPC classification number: A61K39/135 ,  A61K48/00 ,  C12N2770/32111 ,  Y10S930/222

Abstract: 본발명은 O 혈청형-SEA(South East Asia, 동남아) 지역형에대한방어항원이발현되는재조합바이러스및 이를포함하는구제역바이러스백신에관한것으로서, 보다상세하게는 ME-SA(Middle-East South America, 중동-남미) 지역형의 O1 마니사(manisa) 백신바이러스에서구조단백질 P1의코딩유전자를 O 혈청형 SEA 지역형바이러스의것으로교체하여외피항원을뒤바꿈으로써 O 혈청형 SEA 지역형에대한방어항원을발현할수 있는재조합바이러스및 이를포함하는구제역바이러스백신에관한것이다.

Abstract translation: 本发明涉及表达用于口蹄疫O型血清型 - 东南亚（SEA）的保护性抗原的重组病毒，以及包含其的口蹄疫病毒疫苗，更具体地涉及表达 通过在中东南美洲（ME-SA）表型O1躁狂症疫苗病毒代替O血清型-SEA和病毒来逆转抗原，通过具有结构蛋白P1编码基因的O血清型SEA表型的保护性抗原和病毒 包含其的口蹄疫疫苗。