首页|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

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

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  • 国家科技期刊平台
  • NETL
  • NSTL
  • 万方数据
为了建立一种高效检测动物血清中蓝舌病病毒(BTV)抗体的检测方法,以BTV血清群特异性VP7抗原与荧光素酶蛋白相融合表达的标记蛋白(VP7-NLuc)为检测抗原建立了荧光素酶免疫吸附(VP7-NLuc-LISA)方法,用于检测BTV抗体.结果表明:BTV阳性血清能特异性识别VP7-NLuc融合蛋白;监测BTV-1灭活疫苗免疫绵羊血清中BTV抗体动态结果表明,VP7-NLucLISA方法与商品化竞争性ELISA(c-ELISA)试剂盒的检测结果呈高度相关性(R=0.76,P＜0.001);对264份田间血清样本的调查结果表明,建立的VP7-NLuc LISA方法与商品化c-ELISA试剂盒的符合率为98.86％,其相对特异性为99.52％,相对敏感性为96.49％.结论:所建立的VP7-NLuc-LISA方法可作为大规模动物BTV流行病学调查和监测的新方法.
Development of an ultrasensitive detection method for anti-BTV antibody based on the luciferase immunosorbent assay
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.

bluetongue virusVP7 proteinluciferase immunosorbent assaycompetitive ELISA(c-ELISA)serological test

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

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中国农业科学院兰州兽医研究所动物疫病防控全国重点实验室,甘肃兰州 730046

江苏省动物重要疫病与人兽共患病防控协同创新中心,江苏扬州 225009

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

国家重点研发计划项目国家肉牛牦牛产业技术体系专项甘肃省自然科学基金项目甘肃省基础研究创新群体项目甘肃省科技厅项目农业科技创新工程项目

2021YFD1800500CARS-3722JR5RA02922JR5RA02422CX8NA011CAAS-ASTIP-2016-LVRI

2024

10.16656/j.issn.1673-4696.2024.0104

中国兽医科学
中国农业科学院兰州兽医研究所

中国兽医科学

CSTPCD北大核心
影响因子：0.524
ISSN：1673-4696
年,卷(期)：2024.54(7)