中国动物传染病学报Issue(6)：1-5,5.
鹅细小病毒NS1蛋白的原核表达及间接ELISA方法的建立
DEVELOPMENT OF AN INDIRECT ELISA ASSAY USING PROKARYOTICALLY EXPRESSED NS1 PROTEIN OF GOOSE PARVOVIRUS
摘要
Abstract
The NS1 gene of GPV Foshan-2009 was amplified in PCR and cloned into pET32a(+) vector. The recombinant NS1 protein was expressed with induction of IPTG and analyzed in SDS-PAGE and Western blot. The expressed NS1 protein reacted well with the positive Muscovy duck serum against GPV. Then, an indirect ELISA assay was developed using the recombinant NS12 protein. The assay factors were determined using the Checkboard method, including coating NS1 protein at 0.5μg/well, test serum dilution at 1:50 and conjugated antibody HRP dilution at 1:10 000. The assay was optimized with a cut-off value of 0.399. The cross testing demonstrated that the indirect ELISA had no reaction with positive sera against Muscovy duck parvovirus, Dnck virus, Dnck hepatitis virus, Newcastle disease virus and Riemerella anatipestifer. The intra-and inter-coefficients of variabilities were 5%and 10%, respectively, suggesting its reproducibility and repeatability. Clinical testing showed that 92%positive rate was obtained with field samples. In conclusion, this indirect ELISA could be used for serological detection and surveillance of Goose parvovirus.关键词
鹅细小病毒/NS1蛋白/ELISAKey words
Goose parvovirus/NS1 protein/ELISA分类
农业科技引用本文复制引用
孙敏华,董嘉文,李林林,袁建丰,邝瑞欢,胡奇林,张建峰..鹅细小病毒NS1蛋白的原核表达及间接ELISA方法的建立[J].中国动物传染病学报,2014,(6):1-5,5.基金项目
公益性行业（农业）科研专项（201003012）；广东省科技计划项目 （）
