National Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute of Chinese Academy of Agricultural Sciences, No. 427, Maduan Street, Harbin 150001, China.
J Virol Methods. 2010 Feb;163(2):405-9. doi: 10.1016/j.jviromet.2009.11.002. Epub 2009 Nov 11.
The VP3-encoding gene of goose parvovirus (GPV) Ep22 strain was cloned and expressed in Escherichia coli. The GPV VP3-encoding gene was 1605 bp in length, and it encoded a 534 amino acid protein with a predicted molecular mass of 59.9 kDa. The VP3 fusion protein expressed in E. coli was detected by goose and Muscovy duck anti-parvovirus polyclonal sera. In addition, an ELISA (VP3-ELISA) using the VP3 protein as the coating antigen for the detection of antibodies to GPV in geese and antibodies to Muscovy duck parvovirus (MDPV) in Muscovy ducks was developed. Compared to the virus neutralization test, the specificity and sensitivity of the VP3-ELISA was 90.2% and 95.2% for goose sera and 91.8% and 96.7% for Muscovy duck sera, respectively. The VP3-ELISA did not react with the anti-sera to other goose or duck pathogens, indicating that this protein is specific for the reorganization of goose or duck anti-parvovirus antibodies. Cross-reactivity between immunoglobulin G antibodies from geese and Muscovy ducks was also tested, and the results reflected the phylogenetic distance between these two birds when using the ELISA. In conclusion, the VP3-ELISA is a sensitive and specific method for detecting antibodies against GPV or MDPV.
鹅细小病毒(GPV)Ep22 株的 VP3 编码基因在大肠杆菌中被克隆和表达。GPV VP3 编码基因长 1605 个碱基,编码一个 534 个氨基酸的蛋白质,预测分子量为 59.9 kDa。鹅和麝香鸭抗小鹅瘟多克隆血清检测到在大肠杆菌中表达的 VP3 融合蛋白。此外,还建立了一种酶联免疫吸附试验(VP3-ELISA),使用 VP3 蛋白作为包被抗原,用于检测鹅的抗 GPV 抗体和麝香鸭的抗小鹅瘟病毒(MDPV)抗体。与病毒中和试验相比,VP3-ELISA 对鹅血清的特异性和敏感性分别为 90.2%和 95.2%,对麝香鸭血清的特异性和敏感性分别为 91.8%和 96.7%。VP3-ELISA 与其他鹅或鸭病原体的抗血清不发生反应,表明该蛋白特异性重组了鹅或鸭抗小鹅瘟病毒抗体。还测试了鹅和麝香鸭免疫球蛋白 G 抗体之间的交叉反应性,使用 ELISA 时反映了这两种鸟类之间的系统发育距离。总之,VP3-ELISA 是一种检测抗 GPV 或 MDPV 抗体的敏感和特异的方法。