中国兽医科学2026，Vol.56Issue(3)：335-341,7.DOI:10.16656/j.issn.1673-4696.2026.0027
牛结节性皮肤病病毒A33R蛋白的截短表达及间接ELISA抗体检测方法的建立
Truncated expression of lumpy skin disease virus A33R protein and establishment of indirect ELISA antibody detection method
摘要
Abstract
To establish a rapid and accurate ELISA antibody detection method for LSDV,this study referred the LSDV A33R gene sequence from GenBank and then designed a truncated gene with the optimized codons by removing the transmembrane region fragment.The truncated gene was synthesized and cloned into the prokaryotic expression plasmid pET-28a,constructing a recombinant expression plasmid pET-28a-A33R△45-67aa.The A33R△45-67aa protein was induced to express by IPTG and purified,then used as a diagnostic antigen to establish an indirect ELISA antibody detection method.The results showed that the recombinant A33R△45-67aa protein with a molecular weight of 21 kDa was successfully expressed and purified in E.coli,and the established indirect ELISA method exhibited good specificity and sensitivity.Both inter-batch and intra-batch coefficients of variation were below 10％,indicating that the indirect ELISA method had high repeatability and stability.The overall coincidence rate of the test results of this method with the French ID Screen®Capripox double antigen sandwich ELISA kit was 97.7％.The indirect ELISA method can be used for the antibody detection of LSDV and will provide technical support for LSD prevention and control.关键词
牛结节性皮肤病病毒/A33R△45-67aa蛋白/截短表达/间接ELISAKey words
lumpy skin disease virus/A33R△45-67aa protein/truncated expression/indirect ELISA分类
农业科技引用本文复制引用
包燕玲,宋昱庆,户鑫兵,钟匀汝,田占成,苟惠天,关贵全,殷宏,独军政..牛结节性皮肤病病毒A33R蛋白的截短表达及间接ELISA抗体检测方法的建立[J].中国兽医科学,2026,56(3):335-341,7.基金项目
国家重点研发计划项目(2024YFD1800100,2021YFD1800500) （2024YFD1800100,2021YFD1800500）
甘肃省基础研究创新群体项目(22JR5RA024) （22JR5RA024）
国家肉牛牦牛产业技术体系专项(CARS-37) （CARS-37）
中国农业科学院科技创新工程(CAAS-ASTIP-2021-LVRI) （CAAS-ASTIP-2021-LVRI）
