以重组mE2蛋白为抗原建立检测猪瘟病毒抗体间接ELISA方法的研究

以在Ｅ．ｃｏｌｉ高效表达的猪瘟病毒Ｅ２基因主要抗原编码区（ｍＥ２）为抗原，以辣根过氧化物酶（ＨＲＰ）标记的兔抗猪ＩｇＧ为二抗，建立了检测猪瘟病毒抗体的间接ＥＬＩＳＡ方法。经检测筛选出最佳反应条件为：１２μｇ／孔纯化的Ｅ．ｃｏｌｉ表达的ｍＥ２重组蛋白包被酶标板，用１０％的兔血清或马血清进行封闭，以正常Ｅ．ｃｏｌｉ裂解上清液稀释待检血清。试验表明应用重组ｍＥ２蛋白作为诊断ＨＣＶ抗原具有特异性高、抗原易纯化和成本低等特点。

Development of anIndirect ELISA for Detection of Antibodies against Hog Cholera Virus Employing Recombinan4 E2 expressedin E.coli

ed mE2 in 005 mol/L carbonate buffer(pH9.6)andincubated for 4 hour at 37,then over night at 4.After washing,100lof10% normal serum of rabbit or horse was added as blocking agent and incubated for one hour at 37.Before the serum samples were added,they were preincubated with the supermatant of the normal abstracts of E.coli for 1530 min at room temperature.The experiment indicated the supernatant of E.coli could eliminate the nonspecific antibodies in the serum samples and it was not necessary to highly purify the antigen.The results showed the assay is characterized by its specficity,simplicity and economical cost.