畜牧与兽医2017，Vol.49Issue(12)：75-79,5.
小反刍兽疫病毒重组N蛋白抗原制备及间接ELISA检测方法的建立
Preparation of peste des petits ruminants virus (PPRV) nucleocapsid protein and establishment of an indirect ELISA for detecting the serum antibody against PPRV
摘要
Abstract
The full-length N protein coding gene of peste des petits ruminants virus (PPRV) was cloned into pBacPAK9 vector and expressed in insect cells.The expressed protein was characterized by sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDSPAGE) and Western blot analysis using a PPRV specific positive serum.The expressed histidine-tagged fusion protein was purified using affinity Ni-IDA column.The characterization and activity of the purified recombinant protein was analyzed.Furthermore,an indirect ELISA method was developed and 137 serum samples were detected by indirect ELISA and cELISA kit from IDvet.The results showed that the expressed PPRV N protein was approximately 59 ku and could react strongly with the sera from the PPRV infected sheep.In addition,the coincidence rate of the two methods was 95.6％.The results demonstrated that the indirect ELISA established in this study works well in PPR diagnosis.关键词
小反刍兽疫病毒/N蛋白/表达/ELISAKey words
PPRV/N protein/expression/ELISA分类
农业科技引用本文复制引用
冯向辉,陈三民,孔汉金,杨璐,申屠芬琴,田新生,张丽,孙明,陈西钊..小反刍兽疫病毒重组N蛋白抗原制备及间接ELISA检测方法的建立[J].畜牧与兽医,2017,49(12):75-79,5.基金项目
公益性行业科研专项经费(201510017-03) （201510017-03）
