Abstract:
A method for diagnosing duck hepatitis virus type 1 is provided to reduce the diagnosis time, improve convenience of diagnosis, and enhance diagnosis accuracy by performing PCR using a primer set specific to duck hepatitis virus type 1. The method for diagnosing duck hepatitis virus type 1 comprises the steps of: (1) pulverizing the liver tissue infected by duck hepatitis virus and suspending the pulverized liver tissue in PBS(phosphate buffer saline); (2) extracting RNA from the supernatant by using LS reagent; (3) synthesizing cDNA from the extracted RNA by using reverse transcriptase(MMLV); (4) performing PCR using synthesized cDNA as a template using 3D gene-specific primers having the nucleotide sequences of SEQ ID NOs:6 and 7; and (5) subjecting the PCR product to electrophoresis by using 1.5% agarose gel, wherein the 3D gene is derived from duck hepatitis virus type 1 and has the nucleotide sequence selected from SEQ ID NO:1 to SEQ ID NO:5.
Abstract translation:提供一种用于诊断1型鸭肝炎病毒的方法,通过使用特异于1型鸭肝炎病毒的引物组进行PCR,减少诊断时间,提高诊断方便性,提高诊断准确度。1型鸭肝炎病毒诊断方法 包括以下步骤:(1)粉碎鸭肝炎病毒感染的肝组织,并将粉碎的肝组织悬浮于PBS(磷酸盐缓冲盐水)中; (2)使用LS试剂从上清液中提取RNA; (3)使用逆转录酶(MMLV)从提取的RNA合成cDNA; (4)使用具有SEQ ID NO:6和7的核苷酸序列的3D基因特异性引物,使用合成的cDNA作为模板进行PCR; 和(5)使用1.5%琼脂糖凝胶对PCR产物进行电泳,其中3D基因来源于1型鸭型肝炎病毒,并具有选自SEQ ID NO:1至SEQ ID NO:5的核苷酸序列。
Abstract:
A low pathogenic avian influenza viral vaccine for preventing H5 subtype avian influenza viral infection is provided to enhance safety and medicinal efficacy of use, and prevent the infection of H5N1 type high pathogenic avian influenza virus safely. The method for preparing the low pathogenic avian influenza viral vaccine for preventing H5 subtype avian influenza viral infection comprises the steps of: (1) culturing a A/wild bird feces/Korea/CSM-2/02(H5N3) strain in a specific pathogen free egg(SPF egg) for 72 hours and collecting allantoic fluid; (2) measuring the titrate of the virus collected in step(1) through hemagglutination inhibition test; (3) concentrating the virus 10 times by centrifuging the virus at 18,000 rpm for 3 hours; (4) attenuating the virus by treating the concentrated virus with 0.1% formalin at 20 deg.C for 12 hours; and (5) mixing the attenuated virus solution with oil adjuvant ISA70 in a weight ratio of 3:7, wherein the low pathogenic avian influenza viral vaccine contains at least one protein encoded by the gene of SEQ ID NO:2 to SEQ ID NO:9.
Abstract translation:提供用于预防H5亚型禽流感病毒感染的低致病性禽流感病毒疫苗,以提高使用的安全性和药用效力,并安全防止H5N1型高致病性禽流感病毒感染。 制备用于预防H5亚型禽流感病毒感染的低致病性禽流感病毒疫苗的方法包括以下步骤:(1)在特定病原体中培养A /野鸟粪/韩国/ CSM-2/02(H5N3)菌株 免费鸡蛋(SPF蛋)72小时并收集尿囊液; (2)通过血细胞凝集抑制试验测定步骤(1)中收集的病毒的滴度; (3)通过以18,000rpm将病毒离心3小时来浓缩病毒10次; (4)通过用0.1%福尔马林在20℃处理浓缩的病毒12小时来减毒病毒; 和(5)将减毒病毒溶液与油佐剂ISA70以3:7的重量比混合,其中低致病性禽流感病毒疫苗含有由SEQ ID NO:2至SEQ ID NO:2的基因编码的至少一种蛋白质。 9。
Abstract:
조류 인플루엔자 바이러스의 HA 단백질 유래의 HA2 절편의 154번째 아미노산인 아스파라긴(N)에 N-글리칸 생성이 일어나지 않도록 154번째 아미노산인 아스파라긴 또는 156번째 아미노산인 세린(S)이나 트레오닌(T)을 다른 아미노산으로 치환하여 HA2 공통 에피톱에 대한 면역원성을 증가시키는 방법과 이러한 방법을 통해 면역원성이 증가된 재조합 바이러스가 제공된다. 본 발명의 HA2 공통 에피톱 면역원성 향상기술과 HA2 공통 에피톱 고면역원성 바이러스는 인플루엔자 예방을 위한 백신주 개발과 백신주로써 유용하게 이용될 수 있다.
Abstract:
본 발명은 조류 인플루엔자 바이러스 항체의 진단키트 및 이를 이용한 진단방법에 관한 것이다. 보다 구체적으로 본 발명은 조류 인플루엔자 바이러스의 핵단백질(Nucleoprotein)에 대한 단클론 항체가 흡착된 마이크로플레이트에 재조합된 핵단백질 항원이 결합된 인플루엔자 항원 흡착 플레이트 및 조류 인플루엔자 바이러스의 핵단백질에 대한 단클론 항체와 효소가 접합된 효소표지항체를 포함하는 것을 특징으로 하는 조류 인플루엔자 바이러스 항체의 진단키트 및 이를 이용하여 검사시료 내 조류 인플루엔자 바이러스 항체를 측정하는 것을 특징으로 하는 진단방법에 관한 것이다. 조류 인플루엔자, 핵단백질, 단클로 항체, 효소표지항체
Abstract:
본 발명은 오리 간염바이러스 1형(duck hepatitis virus type 1) 유전자의 감별진단방법(differential diagnostic method)에 관한 것으로, 더욱 상세하게는 2C 유전자, 캡시드(capsid) 유전자 및 5'-UTR(untranslated region) 유전자를 표적(target)하는 특이 프라이머를 사용하여 RT-PCR 방법으로 오리 간염바이러스 1형과 유전자형이 다른 오리 간염바이러스 유전자를 감별 진단하는 방법에 관한 것이다. 또한 본 발명에 따른 감별진단방법은 조류 유래의 다른 병원성 RNA 바이러스들과도 신속하고 정확하게 감별적으로 진단하는데 유용하다. 오리 간염바이러스 1형 및 아형, PCR, 5′-UTR 유전자, 캡시트 유전자, 3D 유전자
Abstract:
A diagnostic strip for a H5 type highly pathogenic avian influenza and general avian influenza is provided to conveniently and rapidly diagnose whether a bird is infected by the H5 type highly pathogenic avian influenza and general avian influenza or not from stool in situ without special testing equipment. The diagnostic strip is characterized in that a monoclonal antibody common to general avian influenza virus in which hemaglutinin antigens are H1 to H15 and a monoclonal antibody specific to a highly pathogenic avian influenza in which a hemaglutinin antigen is H5 are used for diagnosing the highly pathogenic and the general avian influenza in which the hemaglutinin antigen is H5 from stool or tissue of chicken. The method for preparing the diagnostic strip comprises the steps of: (a) respectively absorbing the monoclonal antibody specific to the highly pathogenic avian influenza virus in which the hemaglutinin antigen is H5 and the monoclonal antibody common to general avian influenza virus into a membrane; (b) respectively attaching the antibody absorbed membrane obtained from the step(a) to gold particles so as to prepare an antibody-gold conjugate and then mixing it; and (c) precipitating the conjugate obtained from the step(b) in a basic material and then drying it.
Abstract:
A virus strain for preparing a vaccine is provided to have excellent defending effect on low pathogenic avian influenza H9N2 virus strain. And a vaccine is provided to be able to effectively prevent diseases caused by the low pathogenic avian influenza H9N2 recording the high mortality of bird. The low pathogenic avian influenza H9N2 virus strain for preparing vaccine is characterized in that a motif of hemangioblast protein cleavage site amino acid sequence has TSGR and it shows the defending capacity against domestic low pathogenic avian influenza virus. The method for preparing a vaccine for the low pathogenic avian influenza H9N2 virus strain comprises the steps of: (a) inactivating the low pathogenic avian influenza H9N2 virus strain; and (b) mixing the inactivated virus with an adequate excipient.
Abstract:
A recombinant protein of avian influenza virus and a diagnostic composition of avian influenza containing the same protein are provided to mass produce the antigen for diagnosis of avian influenza cheaply, so that the disease caused by avian influenza is prevented. The method for producing the recombinant nucleus protein of avian influenza virus comprises the steps of: (1) amplifying the gene encoding nucleocapsid having the nucleotide sequence of SEQ ID NO:1 as a common protein of MS96 strain with PCR, and cloning the PCR amplified gene into the TA cloning vector; (2) re-isolating the NP gene inserted into the TA cloning vector with a restriction enzyme EcoR I, and inserting the isolated NP gene into the baculovirus transition vector pBacPAK8 to prepare the baculovirus transition vector pBacAINP; (3) co-transfecting the Sf21 cell with the baculovirus and the transition vector pBacAINP to a recombinant baculovirus(rAINP) expressing the avian influenza virus NP protein; and (4) verifying the expression of avian influenza virus NP(nucleus protein) in the Sf21 cell by using a monoclonal antibody specific to the avian influenza virus NP.
Abstract translation:提供禽流感病毒的重组蛋白和含有相同蛋白质的禽流感诊断组合物,以便大量生产用于禽流感诊断的抗原,从而防止禽流感引起的疾病。 制备禽流感病毒重组核蛋白的方法包括以下步骤:(1)用PCR扩增编码具有SEQ ID NO:1的核苷酸序列的核衣壳的基因作为MS96菌株的常见蛋白质,并将PCR扩增 基因进入TA克隆载体; (2)用限制酶EcoR I重新分离插入TA克隆载体的NP基因,将分离的NP基因插入杆状病毒转移载体pBacPAK8中以制备杆状病毒转移载体pBacAINP; (3)将杆状病毒和过渡载体pBacAINP的Sf21细胞共转染至表达禽流感病毒NP蛋白的重组杆状病毒(rAINP); 和(4)通过使用特异于禽流感病毒NP的单克隆抗体来验证Sf21细胞中禽流感病毒NP(核蛋白)的表达。