猪流行性腹泻病毒特异性SIgA间接ELISA检测方法的建立及初步应用

Establishment and preliminary application ofindirect ELISA method for detection of specific SIgA against porcine epidemic diarrhea virus

以乳汁中SIgA特有的成分SC蛋白为研究对象,运用基因克隆、原核表达等方法,制备兔抗猪SC多克隆抗体,使用猪流行性腹泻病毒(porcine epidemic diarrhea virus,PEDV)S截短蛋白进行包被后,初步建立了一种可以检测母猪初乳中PEDV特异性SIgA抗体的ELISA方法。结果显示,PEDV特异性SIgA检测方法具有特异性强、敏感性高、重复性好等特点。结果表明,该方法适用于检测临床中SIgA。该方法的成功建立为生产中评估仔猪从乳汁中获得SIgA的效率及制定更为完善的母猪免疫程序提供了有效的技术支持。

Porcine epidemic diarrhea is caused by porcine epidemic diarrhea virus.Pigs are the only susceptible animals and all ages of pigs are susceptible.The disease has the characteristics of acute onset,rapid transmission and high mortality in the clinic.Some studies indicate that PEDV has an immune pathway of“intestinal-mammary-sIgA axis”.In order to better prevent and control porcine epidemic diarrhea,a more complete immune program is formulated in production practice according to the efficiency of passive immunity obtained by piglets from milk.In this study,SC protein,a unique component of SIgA in breast milk was cloned and prokaryotic expressed to prepare rabbit anti-pig SC polyclonal antibodies.Coated with PEDV spike proteins,an ELISA method was established to detect PEDV-specific SIgA antibodies in sow colostrum.The results showed that the porcine epidemic diarrhea virus-specific SIgA detection method had the characteristics of strong specificity,high sensitivity and good reproducibility,indicating that the method is suitable for the detection of SIgA in clinical samples.The successful establishment of the ELISA method provides effective technical support for evaluating the efficiency of piglets in obtaining SIgA from milk and developing a more complete sow immunization program during production.