CAJ | 학술논문

研究以纯化的羊抗草鱼呼肠孤病毒多克隆抗体作为捕获抗体，抗草鱼呼肠孤病毒的单克隆抗体为检测抗体，建立草鱼呼肠孤病毒抗原捕获ELISA检测方法，其最佳反应条件是：羊多克隆抗体包埋浓度为2.5μg/mL，抗草鱼呼肠孤病毒单克隆抗体工作浓度为1：400稀释，以3％牛血清白蛋白作为封闭液。该方法的检测限为5×105 pfu/mL，且能够特异性检出发病草鱼内脏组织中的草鱼呼肠孤病毒。特异性试验结果表明该方法具有较高的特异性，与病毒性出血性败血症病毒、传染性造血器官坏死病毒和鲤春血症病毒等水产动物病毒株均不反应。该方法还具有较好的稳定性。
연구이순화적양항초어호장고병독다극륭항체작위포획항체，항초어호장고병독적단극륭항체위검측항체，건립초어호장고병독항원포획ELISA검측방법，기최가반응조건시：양다극륭항체포매농도위2.5μg/mL，항초어호장고병독단극륭항체공작농도위1：400희석，이3％우혈청백단백작위봉폐액。해방법적검측한위5×105 pfu/mL，차능구특이성검출발병초어내장조직중적초어호장고병독。특이성시험결과표명해방법구유교고적특이성，여병독성출혈성패혈증병독、전염성조혈기관배사병독화리춘혈증병독등수산동물병독주균불반응。해방법환구유교호적은정성。
An antigen-capture enzyme-linked immunoassay(ELISA)was developed for the detection of grass carp reovirus(GCRV)with puriifed polyclonal antiserum against GCRV as capture antibody and monoclonal antibodies against GCRV as detection antibody. The 96-well enzyme immunoassay(EIA)plates were coated with 100μL puri-ifed polyclonal antiserum at concentration of 2.5μg/mL and blocked with 3%?bovine serum albumin(BSA). The working concentration of monoclonal antibody was diluted to 1:400 with distilled water. GCRV was detected in culture supernatants(5×105 pfu/mL)and in the extracts of kidney and spleen of infected grass carp. The assay was highly speciifc:viral hemorrhagic septicemia virus,infectious haematopoietic necrosis virus,spring viraemia of carp virus, epizootic haematopoietic necrosis virus,chinook salmon reovirus could not be detected by the developed ELISA and the method was also of good stability.