猪流行性腹泻IgA抗体间接ELISA检测方法的建立  

Development of an indirect ELISA method for detection of IgA antibody against porcine epidemic diarrhea virus

作　　者：丁振江[1] 库旭钢[1] 闫贵伟 陈淑华[1] 何启盖[1] 

机构地区：[1]农业微生物学国家重点实验室华中农业大学动物医学院,湖北武汉430070

出　　处：《畜牧与兽医》2014年第11期1-5,共5页Animal Husbandry & Veterinary Medicine

基　　金：国家自然科学基金(31272572)

摘　　要：将猪流行性腹泻病毒(PEDV)pET-32a-S1D重组表达质粒转化至大肠杆菌BL-2l(DE3)感受态细胞,并成功表达目的蛋白S1D。以纯化后蛋白作为抗原,建立了针对乳汁中PEDV的IgA抗体间接ELISA检测方法。确定最佳抗原包被浓度为1μg/mL;乳汁最佳稀释度为1∶10。该检测方法敏感性高,可重复性好。该研究建立的间接ELISA方法为母猪乳汁中猪流行性腹泻病毒IgA抗体的检测提供了一种快速简便的诊断方法。The fusion expression plasmid pET- 32a- S1 D of porcine epidemic diarrhea virus（PEDV） was transformed into Escherichia coli strain BL21（DE3）,and then the target protein S1 D of PEDV was successfully expressed after IPTG induction. Based on the purified recombinant protein,a novel indirect ELISA method was developed to detect IgA antibody against PEDV in sow milk. The optimized reaction conditions were as follows： antigen working concentration was 1μg /mL; milk sample was diluted at 1∶ 10. This method has high sensitivity and good repeatability. The indirect ELISA method established in this study will provide a convenient tool for the investigation of milk IgA level among pig herds.

关 键 词：猪流行性腹泻病毒 S1D蛋白 乳汁 IGA 间接ELISA 

分 类 号：S851.34[农业科学—预防兽医学]