Abstract:
본 발명은 웨스트나일 바이러스에 대한 중화항체 검출방법 및 웨스트나일병의 진단을 위한 정보제공방법에 관한 것이다. 본 발명의 웨스트나일 바이러스에 대한 중화항체의 검출방법을 이용할 경우, 차폐시설 내 작업 시간을 단축하여 고 위험 병원체에 노출될 수 있는 작업자의 위험성을 줄이고, 실험 절차를 간소화하여 포르말린 등과 같은 유독 폐기물 사용을 절감할 수 있을 뿐 아니라, 숙련자가 아닌 실험자도 쉽게 웨스트나일 바이러스에 대한 중화항체 여부를 판정하고, 한 번에 대량의 시료 처리가 가능하다.
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。
Abstract:
PURPOSE: A risk of operator exposed to high risk pathogen detecting method of neutralizing antibody about West Nile virus shortens occlusion facility inside working hours is reduced. CONSTITUTION: A detecting method of neutralizing antibody about West Nile virus comprises the steps of: (a) A step of reacting the West Nile virus and specimen in the neutralization test culture medium of the West Nile virus consisting of the DMEM (Dulbeco's modified Eagle's medium) culture medium including the glucose of 8 through 12 g / L; (b) A feeling verse writing key in the West Nile virus sensibility cell is the step the reactant of (a) step; (c) A step that removes the supernatant of the reactant of (b) step and cultivated in the West Nile virus neutralization test culture medium of (a) step; (d) A step of measuring the absorbance of the culture medium of (c) step are included.
Abstract:
본 발명은 웨스트나일바이러스와 일본뇌염바이러스의 감별 진단에 관한 것으로, 보다 자세하게는 웨스트나일바이러스와 일본뇌염바이러스 각각에 특이적인 프라이머 세트, 상기 프라이머 세트를 포함하는 웨스트나일바이러스와 일본뇌염바이러스의 감별 검출용 조성물 및 상기 프라이머 세트를 이용하여 RT-PCR을 실시하는 단계를 포함하는 웨스트나일바이러스와 일본뇌염바이러스의 감별 검출 방법에 관한 것이다. 상기한 2종의 바이러스들은 모두 플라비바이러스속에 속하는 유사바이러스로서 사람과 말 등에 일단 감염되면 뇌염과 고열 등의 임상증상이 유사하여 감별진단이 필요하다. 본 발명은 실험실 진단시 단일튜브에서 상기한 2종의 플라비바이러스들을 동시에 검사할 수 있기 때문에 신속한 진단에 활용이 가능하다.
Abstract:
PURPOSE: A method for distinguishing and diagnosing west nile virus and Japanese encephalitis virus is provided to simultaneously test two kinds of Togaviridae and flaviviridae viruses. CONSTITUTION: A primer set for detecting west nile virus and Japanese encephalitis virus contains: a west nile-specific primer set having a sense primer of sequence numbers 1, 2, 3, and 4 and an antisense primer of sequence numbers 5, 6, 7, 8, and 9; and a Japanese encephalitis virus-specific primer set having a sense primer of sequence numbers 10 and 11 and an antisense primer of sequence numbers 12 and 13. A composition for detecting the west nile virus and Japanese encephalitis virus contains the primer sets. The composition is an RT-PCR kit.
Abstract:
PURPOSE: A single tube-multiplex RT-PCR for simultaneously detecting eastern equine encephalitis, western equine encephalitis, and venezuelan equine encephalitis viruses is provided to construct quick and efficient virus diagnosis system. CONSTITUTION: A single tube-multiplex RT-PCR for simultaneously detecting eastern equine encephalitis, western equine encephalitis, and venezuelan equine encephalitis viruses contains RT-PCR primer set, reaction buffer. DNA polymerase, and reverse transcriptase. A RT-PCR primer set contains primers of sequence numbers 1 and 2, sequence numbers 3 and 4, or sequence number 5 and 6.