中国兽医科学2024，Vol.54Issue(7)：881-887,7.DOI:10.16656/j.issn.1673-4696.2024.0104
荧光素酶免疫吸附法高灵敏检测蓝舌病病毒抗体方法的建立
Development of an ultrasensitive detection method for anti-BTV antibody based on the luciferase immunosorbent assay
摘要
Abstract
To establish an efficient method for detecting anti-bluetongue virus(BTV)antibody in animal serum,BTV VP7 protein fused with a reporter Nano-luciferase(VP7-NLuc LISA)as the diagnostic antigen was used for the development of a luciferase immunosorbent assay(LISA).Results showed that BTV-positive serum could specifically recognize VP7-NLuc fused protein.The dynamic monitoring of BTV antibody in serum of sheep immunized with BTV-1 inactivated vaccine showed that the detection results of VP7-NLuc LISA method and commercial competitive ELISA(c-ELISA)kit were highly correlated(R=0.76,P＜0.001).In investigating 264 field serum samples,the relative specificity and sensitivity of the VP7-NLuc LISA were 99.52％and 96.49％,respectively,which was 98.86％in agreement with the commercial c-ELISA kit.In conclusion,the established VP7-NLuc-LISA method can serve as a new method for large-scale animal BTV epidemiological investigation and monitoring.关键词
蓝舌病病毒/VP7蛋白/荧光素酶免疫吸附试验/c-ELISA/血清学检测Key words
bluetongue virus/VP7 protein/luciferase immunosorbent assay/competitive ELISA(c-ELISA)/serological test分类
农业科技引用本文复制引用
张鸿歌,田占成,独军政,王琼洁,康棣,户鑫兵,张忠辉,关贵全,殷宏..荧光素酶免疫吸附法高灵敏检测蓝舌病病毒抗体方法的建立[J].中国兽医科学,2024,54(7):881-887,7.基金项目
国家重点研发计划项目(2021YFD1800500) （2021YFD1800500）
国家肉牛牦牛产业技术体系专项(CARS-37) （CARS-37）
甘肃省自然科学基金项目(22JR5RA029) （22JR5RA029）
甘肃省基础研究创新群体项目(22JR5RA024) （22JR5RA024）
甘肃省科技厅项目(22CX8NA011) （22CX8NA011）
农业科技创新工程项目(CAAS-ASTIP-2016-LVRI) （CAAS-ASTIP-2016-LVRI）
