公开(公告)号：KR1020040051199A

公开(公告)日：2004-06-18

申请号：KR1020020079085

申请日：2002-12-12

Applicant: 대한민국(농림축산식품부 농림축산검역본부장)

Inventor： 송재영 ,  최은진 ,  박종현 ,  안동준 ,  차상호 ,  권준헌

IPC: G01N33/543

Abstract: PURPOSE: A detection kit for swine cholera virus antibody using immunochromatography method is provided, thereby rapidly detecting the swine cholera virus antibody, and enabling the in-situ detection of the swine cholera virus antibody, so that the swine cholera virus antibody can be rapidly detected in farmhouses. CONSTITUTION: The detection kit for swine cholera virus antibody using immunochromatography method is prepared by binding a protein antigen(1), preferably E2 protein of swine cholera virus to strip type nitrocellulose membrane(5); and binding anti-swine IgG antibody(2) to the opposite side of the nitrocellulose membrane(5). The method for detecting the swine cholera virus antibody comprises the steps of: contacting a swine blood sample with the nitrocellulose membrane(5) by spotting the sample on the sample pad through a sample loading hole(4); transferring the swine blood sample to the swine cholera virus antigen(1) via the nitrocellulose membrane(5), and transferring the anti-swine IgG gold conjugate adsorbed in a conjugate pad to the absorbing pad(3); and measuring the occurrence of red purple by the antigen-antibody reaction followed by the gold particle reaction with the antigen-antibody conjugate in the antigen fixed region(1).

Abstract translation: 目的：提供使用免疫色谱法的猪霍乱病毒抗体检测试剂盒，从而快速检测猪霍乱病毒抗体，并能够对猪霍乱病毒抗体进行原位检测，从而快速检测猪霍乱病毒抗体 在农舍 构成：使用免疫色谱法测定猪霍乱病毒抗体检测试剂盒是通过将蛋白质抗原（1），优选鸡蛋霍夫病毒E2蛋白与带状硝酸纤维素膜结合而制备的（5）; 并将抗猪IgG抗体（2）与硝酸纤维素膜（5）的相反侧结合。 检测猪霍乱病毒抗体的方法包括以下步骤：通过样品装载孔（4）将样品垫上的样品点样，使猪血样与硝酸纤维素膜接触（5）; 通过硝酸纤维素膜（5）将猪血液样品转移到猪霍乱病毒抗原（1），并将吸附在共轭垫中的抗猪IgG金缀合物转移到吸收垫（3）上; 并通过抗原 - 抗体反应随后与抗原固定区域（1）中的抗原 - 抗体缀合物进行金颗粒反应来测量红紫色的发生。

公开(公告)号：KR1020040047484A

公开(公告)日：2004-06-05

申请号：KR1020020075721

申请日：2002-11-30

Applicant: 대한민국(농림축산식품부 농림축산검역본부장)

Inventor： 차상호 ,  박종현 ,  송재영 ,  안동준 ,  최은진 ,  김종염 ,  권준헌

IPC: C12N15/861

CPC classification number: C12N15/861 ,  C07K14/005 ,  C12N2770/24034

Abstract: PURPOSE: Recombinant porcine adenovirus type 3 expressing Japanese encephalitis virus gene NS1 and use thereof are provided, thereby inducing immunity to Japanese encephalitis virus in human, and improving stability of the vaccine by using adenovirus. CONSTITUTION: A recombinant vector of porcine adenovirus type 3 expressing Japanese encephalitis virus gene NS1, pPAV3E3-NS1(KCTC 10377BP), is prepared by removing E3 gene of porcine adenovirus type 3, and inserting the Japanese encephalitis virus gene NS1 under the control of E3 promoter, wherein the Japanese encephalitis virus gene NS1 has the nucleotide sequence set forth in SEQ ID NO: 1. A host cell transformed with the recombinant vector pPAV3E3-NS1(KCTC 10377BP) is provided. The recombinant porcine adenovirus type 3 expressing Japanese encephalitis virus gene NS1(PAV3E3-NS1) is obtained by culturing the transformed host cell. A vaccine for Japanese encephalitis virus comprises the recombinant porcine adenovirus type 3 expressing Japanese encephalitis virus gene NS1.

Abstract translation: 目的：提供3型表达日本脑炎病毒基因NS1的重组猪腺病毒及其用途，从而诱导人体对日本脑炎病毒的免疫，并通过使用腺病毒提高疫苗的稳定性。 构成：通过除去3型猪腺病毒的E3基因，并将日本脑炎病毒基因NS1插入到E3的控制下，制备3型表达日本脑炎病毒基因NS1，pPAV3E3-NS1（KCTC 10377BP）的猪腺病毒重组载体 启动子，其中日本脑炎病毒基因NS1具有SEQ ID NO：1所示的核苷酸序列。提供用重组载体pPAV3E3-NS1（KCTC 10377BP）转化的宿主细胞。 通过培养转化的宿主细胞获得3型表达日本脑炎病毒基因NS1（PAV3E3-NS1）的猪腺病毒。 日本脑炎病毒疫苗包含3型表达日本脑炎病毒基因NS1的重组猪腺病毒。

公开(公告)号：KR101592247B1

公开(公告)日：2016-02-05

申请号：KR1020120039157

申请日：2012-04-16

Applicant: 바이오코아 주식회사 ,  건국대학교 산학협력단 ,  대한민국(농림축산식품부 농림축산검역본부장)

Inventor： 황승용 ,  김지훈 ,  이원선 ,  송창선 ,  이동훈 ,  이윤정 ,  강현미 ,  최준구 ,  권준헌 ,  박최규

IPC: C12N15/11 ,  C12Q1/68 ,  C12Q1/04

Abstract: 본발명은조류인플루엔자바이러스의혈청형을결정하는 HA 유전자의아형 (H5형, H7형, H9형) 및 NA 유전자의아형(N1형, N2형, N3형) 판별과병원성의검사가동시에가능하면서도조류인플루엔자바이러스를조기에신속하고정확하게검사할수 있는조류인플루엔자바이러스검사용프로브조성물, 유전자칩 그리고이를이용한조류인플루엔자바이러스검사방법을제공한다. 본발명에따르면, 저가의비용으로 5시간이내에고병원성조류인플루엔자바이러스의혈청형및 병원성감별이가능하고저병원성 H9형바이러스의감별과더불어경제적이며빠른진단으로고병원성조류인플루엔자발생시조기진단에따른초동대처로질병발생피해를최소화하는데기여할수 있다.

公开(公告)号：KR101491817B1

公开(公告)日：2015-02-16

申请号：KR1020120150890

申请日：2012-12-21

Applicant: 대한민국(농림축산식품부 농림축산검역본부장)

Inventor： 최강석 ,  계수정 ,  김지예 ,  권준헌 ,  이희수

IPC: C12N15/63 ,  C12N15/45 ,  C12N15/85 ,  G01N33/68

Abstract: 본 발명은 제8형 조류파라믹소바이러스 (avian paramyxovirus-8, APMV-8)의 헤마글루티닌-뉴라미니다아제 (hemagglutinin-neuraminidase, HN) 단백질을 코딩하는 유전자를 포함하는 재조합 바이러스 발현 벡터, 상기 벡터에 의해 형질전환된 제8형 조류파라믹소바이러스의 헤마글루티닌-뉴라미니다아제 단백질을 발현하는 재조합 곤충 세포, 상기 재조합 곤충세포가 발현하는 제8형 조류파라믹소바이러스의 헤마글루티닌-뉴라미니다아제 재조합 항원 단백질을 포함하는 제8형 조류파라믹소바이러스 진단용 조성물, 진단 키트 및 이를 이용한 진단 방법에 관한 것이다. 본 발명에 따른 진단용 조성물은 살아있는 바이러스 취급에 따른 오염 가능성 없이 안전하고, 신속하게 대량의 샘플로부터 제8형 조류파라믹소바이러스의 감염 여부를 정확하게 진단할 수 있는 우수한 효과를 가지고 있다.

公开(公告)号：KR101463001B1

公开(公告)日：2014-12-04

申请号：KR1020120150870

申请日：2012-12-21

Applicant: 대한민국(농림축산식품부 농림축산검역본부장)

Inventor： 최강석 ,  계수정 ,  김지예 ,  권준헌 ,  이희수 ,  성환우

IPC: C12N15/63 ,  C12N15/45 ,  C12N15/85 ,  C07K14/08

Abstract: 본발명은베큘러바이러스에의하여발현된조류파라믹소바이러스-7 재조합 HN 단백질및 이를이용한조류파라믹소바이러스-7 진단방법에관한것이다. 본발명에따른 APMV-7 HN 단백질을발현하는베큘러바이러스또는이에의하여형질감염된곤충세포는 APMV-7 HN 단백질을고농도로생산하며, 이를이용하여세포배양이가능한일반실험실에서도손쉽게대량의항원을제조할수 있고, 곤충세포배양법으로일주일이내에항원진단액제조가가능하므로항원진단액제조기간을최소일주일이상단축할수 있는효과가있다. 또한, 본발명에의해제조한 APMV-7 HN 단백질항원은닭 적혈구에대한혈구응집능과열 안정성이있고, APMV-7 면역혈청에의해 HI 반응이특이적으로나타났을뿐만아니라, 종래의항원 (APMV-7 바이러스항원)과 HI반응법검사결과와비교하였을때 동일한검사결과를나타내는효과가있다. 따라서, 감염성 APMV-7 바이러스를확보하고있지않아도종래의 APMV-7 바이러스항원진단액을본 발명에의해제조한 APMV-7 HN 단백질항원으로대체하여사용할수 있다.

公开(公告)号：KR1020140081374A

公开(公告)日：2014-07-01

申请号：KR1020120151046

申请日：2012-12-21

Applicant: 대한민국(농림축산식품부 농림축산검역본부장)

Inventor： 최강석 ,  계수정 ,  김지예 ,  권준헌 ,  이희수 ,  성환우

IPC: C12N15/63 ,  C12N15/45 ,  C12N15/85 ,  G01N33/68

Abstract: The present invention relates to avian paramyxovirus-9 (APMV-9) recombinant hemagglutinin-neuraminidase (NH) protein expressed by baculoviruses, and a diagnostic method of APMV-9 using the same. The baculoviruses expressing APMV-9 HN protein according to the present invention, or insect cells transfected with the baculoviruses produce a high concentration of APMV-9 HN protein; easily enable the production of a large number of antigens even in a general laboratory allowing cell culture by using the APMV-9 HN protein; and enable the production of an antigen diagnostic reagent within a week by an insect cell culture method, thereby having an effect of shortening the production period of the antigen diagnostic reagent by at least one week. In addition, an APMV-9 HN protein antigen produced by the present invention has hemagglutination ability for erythrocytes of a chicken and thermal stability; has exhibited a specific HI reaction by APMV-9 immune serum; and also has an effect of exhibiting the same test result in comparison with a HI reaction test result from a conventional antigen (APMV-9 virus antigen). Therefore, a conventional APMV-9 virus antigen diagnostic reagent can be replaced with the APMV-9 HN protein antigen produced by the present invention even if infectious APMV-9 viruses are not secured.

Abstract translation: 本发明涉及由杆状病毒表达的禽副粘液病毒-9（APMV-9）重组血凝素 - 神经氨酸酶（NH）蛋白，以及使用其的APMV-9的诊断方法。 表达本发明的APMV-9HH蛋白的杆状病毒或用杆状病毒转染的昆虫细胞产生高浓度的APMV-9HH蛋白; 即使在通常使用APMV-9HH蛋白的细胞培养的实验室中也容易产生大量抗原; 并且能够通过昆虫细胞培养法在一周内生产抗原诊断试剂，从而具有将抗原诊断试剂的生产周期缩短至少一周的效果。 此外，本发明生产的APMV-9HH蛋白抗原具有鸡的红细胞的血细胞凝集能力和热稳定性; 已经通过APMV-9免疫血清显示出特异性的HI反应; 并且与来自常规抗原（APMV-9病毒抗原）的HI反应试验结果相比，具有显示相同试验结果的效果。 因此，即使不能确保感染性APMV-9病毒，也可以用本发明生产的APMV-9HH蛋白抗原代替传统的APMV-9病毒抗原诊断试剂。

公开(公告)号：KR1020140081337A

公开(公告)日：2014-07-01

申请号：KR1020120150976

申请日：2012-12-21

Applicant: 대한민국(농림축산식품부 농림축산검역본부장)

Inventor： 최강석 ,  계수정 ,  김지예 ,  권준헌 ,  이희수 ,  성환우

IPC: C12N15/63 ,  C12N15/45 ,  C12N15/85 ,  G01N33/68

Abstract: The present invention relates to a recombinant virus expression vector including a gene which encodes hemagglutinin-neuraminidase (HN) protein of avian paramyxovirus-3 (APMV-3); to recombinant insect cells which transformed by the vector and expressing the HN protein of APMV-3; and to a diagnostic composition for APMV-3 including the recombinant antigenic HN protein of APMV-3 expressed by the recombinant insect cells, a diagnostic kit including the HN protein, and a diagnostic method using the HN protein. The diagnostic composition for APMV-3 according to the present invention is safe without the possibility of contamination caused by treatment of live viruses and has an excellent effect of quickly and accurately diagnosing whether there is infection with APMV-3 from a large number of samples.

Abstract translation: 本发明涉及包含编码禽副粘病毒-3（APMV-3）的血凝素 - 神经氨酸酶（HN）蛋白的基因的重组病毒表达载体; 通过载体转化并表达APMV-3的HN蛋白的重组昆虫细胞; 以及包含由重组昆虫细胞表达的APMV-3的重组抗原性HN蛋白质的APMV-3诊断组合物，包含HN蛋白质的诊断试剂盒和使用HN蛋白质的诊断方法。 根据本发明的APMV-3的诊断组合物是安全的，没有由病毒治疗引起的污染的可能性，并且具有快速且准确地诊断来自大量样品是否存在APMV-3感染的优异效果。

公开(公告)号：KR1020140081294A

公开(公告)日：2014-07-01

申请号：KR1020120150890

申请日：2012-12-21

Applicant: 대한민국(농림축산식품부 농림축산검역본부장)

Inventor： 최강석 ,  계수정 ,  김지예 ,  권준헌 ,  이희수

IPC: C12N15/63 ,  C12N15/45 ,  C12N15/85 ,  G01N33/68

Abstract: The present invention relates to a recombinant virus expression vector including a gene which encodes hemagglutinin-neuraminidase (HN) protein of avian paramyxovirus-8 (APMV-8); to recombinant insect cells transformed by the vector and expressing the HN protein of APMV-8; and to a diagnostic composition for APMV-8 including the recombinant antigenic HN protein of APMV-8 expressed by the recombinant insect cells, a diagnostic kit including the HN protein, and a diagnostic method using the HN protein. The diagnostic composition for APMV-8 according to the present invention is safe without the possibility of contamination caused by treatment of live viruses and has an excellent effect of quickly and accurately diagnosing whether there is infection with APMV-8 from a large number of samples.

Abstract translation: 本发明涉及包含编码禽副粘病毒-8（APMV-8）的血凝素 - 神经氨酸酶（HN）蛋白的基因的重组病毒表达载体; 通过载体转化并表达APMV-8的HN蛋白的重组昆虫细胞; 以及包含由重组昆虫细胞表达的APMV-8的重组抗原性HN蛋白质的APMV-8的诊断组合物，包含HN蛋白质的诊断试剂盒和使用HN蛋白质的诊断方法。 根据本发明的APMV-8的诊断组合物是安全的，没有由病毒治疗引起的污染的可能性，并且具有快速且准确地诊断来自大量样品是否存在APMV-8感染的优异效果。

公开(公告)号：KR101374192B1

公开(公告)日：2014-03-14

申请号：KR1020110139058

申请日：2011-12-21

Applicant: 대한민국(농림축산식품부 농림축산검역본부장)

Inventor： 최강석 ,  전우진 ,  이은경 ,  계수정 ,  김새로미 ,  박미자 ,  권준헌

IPC: C12N15/866 ,  C12N15/66 ,  C12Q1/68 ,  C07K14/08

Abstract: 본 발명은 제4형 조류파라믹소바이러스(avian paramyxovirus-4, APMV-4)의 진단에 관한 것이다
상기와 같은 본 발명에 따르면, 제4형 조류파라믹소바이러스(avian paramyxovirus-4, APMV-4) HN(haemagglutinin-neuraminidase) 유전자를 포함하는 재조합 베큘로바이러스 발현벡터를 곤충세포에 형질감염시키고, 상기 곤충세포에서 발현된 APMV-4 HN 단백질 및 이를 포함하는 혈구응집억제시험법(hemagglutination inhibition, HI법) 검사용 NDV 항원진단액과 상기 NDV 항원진단액을 이용한 혈구응집억제시험법(hemagglutination inhibition, HI법) 검사용 APMV-4 항체진단액을 제공함으로서, 살아있는 APMV-4를 취급하는 과정에서 발생할 수 있는 주변 오염이나 질병 전염의 위험성을 배제할 뿐만 아니라 신속하고 경제적인 방법으로 APMV-4 항원 및 항체진단액을 대량생산할 수 있는 효과가 있다.

公开(公告)号：KR1020130085557A

公开(公告)日：2013-07-30

申请号：KR1020110139058

申请日：2011-12-21

Applicant: 대한민국(농림축산식품부 농림축산검역본부장)

Inventor： 최강석 ,  전우진 ,  이은경 ,  계수정 ,  김새로미 ,  박미자 ,  권준헌

IPC: C12N15/866 ,  C12N15/66 ,  C12Q1/68 ,  C07K14/08

CPC classification number: C12N15/866 ,  C07K14/08 ,  C12N15/66 ,  C12Q2525/131 ,  C12Q2531/113

Abstract: PURPOSE: HN protein of avian paramyxovirus-4 (APMV-4) and an antigen diagnostic solution for HI method examination including the same are provided to be able to exclude the danger of neighbor contamination or disease contagion in the process of treating live APMV-4, and to be able to mass-produce the APMV-4 antigen and antibody diagnostic solution in a rapid and economic method. CONSTITUTION: A manufacturing method of an expression vector of recombinant baculovirus including HN gene of APMV-4 comprises (1) a step of extracting virus genome RNA from APMV-4 La Sota strain; (2) a step of synthesizing cDNA of HN protein gene of APMV-4 by using the extracted RNA and reverse primer of Sequence 2; (3) a step of amplifying the synthesized cDNA, forward primer of Sequence 1, and reverse primer of Sequence 2 by using PCR, and synthesizing DNA; (4) a step of extracting DNA fragment by treating the synthesized DNA with EcoR1 and Hind III; and (5) a step of inserting the extracted DNA fragment into the multicloning site of pFastBac^TM1 expression vector having baculovirus polyhedrin promoter (PPH). The forward primer of Sequence 1 is that EcoR1 restriction enzyme acting site is inserted into the upper part of ATG of 5' end terminal. The revers primer of Sequence 2 is that Hind III restriction enzyme acting site is inserted into the upper part of end codon of 5' end terminal.

Abstract translation: 目的：提供禽副粘病毒-4（APMV-4）的HN蛋白和包括其中的HI方法检测的抗原诊断溶液，以能够排除在治疗APMV-4过程中相邻污染或疾病传染的危险 并能够以快速，经济的方式大量生产APMV-4抗原和抗体诊断溶液。 构成：APMV​​-4的HN基因重组杆状病毒表达载体的制造方法包括：（1）从APMV-4 La Sota菌株中提取病毒基因组RNA的步骤; （2）使用提取的RNA和序列2的反向引物合成APMV-4的HN蛋白基因的cDNA的步骤; （3）使用PCR扩增合成的cDNA，序列1的正向引物和序列2的反向引物的步骤，合成DNA; （4）通过用EcoRⅠ和HindⅢ处理合成的DNA来提取DNA片段的步骤; （5）将提取的DNA片段插入具有杆状病毒多角体蛋白启动子（PPH）的pFastBac TM TM1表达载体的多克隆位点的步骤。 序列1的正向引物是EcoR1限制酶作用位点插入5'端末端ATG的上部。 序列2的反向引物是HindIII限制酶作用位点插入5'末端末端密码子的上部。