中国动物传染病学报2016，Vol.24Issue(2)：20-24,5.
双抗夹心ELISA检测兔出血症病毒抗原方法的建立及初步评估
DEVELOPMENT AND PRELIMINARY EVALUATION OF A DOUBLE ANTIBODY SANDWICH ELISA FOR DETECTING RABBIT HEMORRHAGIC DISEASE VIRUS ANTIGENS
摘要
Abstract
In this study, a rapid, specific double antibody sandwich ELISA (DAS-ELISA) method for detection of Rabbit hemorrhagic disease virus (RHDV) was developed. By using the pairing of monoclonal antibodies, it was confirmed that the method was developed using anti-VP60 monoclonal antibody 9H9 as capturing antibody and HRP-9H9 as detection antibody. The results showed that the optimal concentration of 9H9 was 0.5μg/mL. The optimal concentration of HRP-9H9 was 5μg/mL. The optimum condition of coating was 37℃ and then 4℃ overnight. The optimal blocking condition was PBST containing 1% gelatin and incubation at 37℃ for 2 h. The best diluent of detecting antibodies was PBST containing 10%FBS. The ELISA method had no cross-reactivity with other caliciviruses. In clinical tests, 85 samples isolated from rabbits were subjected to DAS-ELISA and RT-PCR. The results show that the DAS-ELISA method had a high level of specificity and sensibility could be used for rapid detection of RHD.关键词
兔出血症病毒/单克隆抗体/双抗夹心ELISAKey words
Rabbit hemorrhagic disease virus/monoclonal antibody/DAS-ELISA分类
农业科技引用本文复制引用
郭慧敏,谭永贵,缪秋红,朱杰,刘腾,陈宗艳,李传峰,刘光清..双抗夹心ELISA检测兔出血症病毒抗原方法的建立及初步评估[J].中国动物传染病学报,2016,24(2):20-24,5.基金项目
公益性农业科研专项(201303046) （201303046）
上海市科委创新项目(13391901602) （13391901602）
