减蛋综合征病毒抗体胶体金免疫层析试纸条的研制及初步应用

Development and Preliminary Application of Colloidal Gold Immunochromatography Test Strips for Egg Drop Syndrome Virus Antibodies

为建立快速检测减蛋综合征病毒(EDSV)抗体的方法,基于EDSV的Fiber蛋白,制备快速检测EDSV抗体的免疫层析试纸条,为养禽业提供针对病原微生物的感染预警及免疫评估。利用胶体金标记EDSV Fiber蛋白抗原为探针,将羊抗鸡IgG和抗Fiber蛋白的抗体包被到硝酸纤维膜上,分别作为质控线和检测线,对家禽EDSV抗体水平进行监测。结果表明,制备的EDSV免疫层析试纸条具有高度特异性,与其他家禽病毒抗体无交叉反应,仅与EDSV抗体反应,在试纸条上产生可见的检测线;并且制备的试纸条具有高度稳定性,在室温条件下可至少保存6个月。应用制备的检测EDSV抗体的免疫层析试纸条对从河南、安徽、山东等地养鸡场采集的576份鸡血清样本检测,与血凝抑制试验(HI)结果进行平行比较,二者的Kappa值为0.878,具有良好一致性。综上,制备的检测ESDV抗体的免疫层析试纸条具有高度的特异性、敏感性、稳定性和易操作性,对EDSV抗体的临床检测具有实际应用价值。

In order to establish a method for rapid detection of egg drop syndrome virus(EDSV)antibody,an immunochromatographic test strip for rapid detection of EDSV antibody was prepared based on the EDSV Fiber protein,which could provide infection warning and immune evaluation for pathogenic microorganisms in poultry industry.Colloidal gold-labeled EDSV Fiber protein was used as a probe,the goat anti-chicken IgG and anti-Fiber polyclonal antibody were blotted onto the nitrocellulose membrane as quality control lines and detection lines respectively to test EDSV antibody.The results showed that the EDSV immunochromatographic test strip was highly specific,had no cross-reactivity with other viral antibodies,it only reacted with EDSV antibodies and produced a visible detection line on the test strip;and the test strips were highly stable and can be stored for at least 6 months at room temperature.The clinical application effect of the strips was tested by detecting EDSV antibodies in 576 serum samples collected from chickens farms in Henan,Anhui and Shandong,and compared with the hemagglutination inhibition test(HI)in parallel,and the Kappa value of the two was 0.878,which was in good agreement.In summary,the immunochromatographic test strips developed in this study are highly specific,sensitive,stable and easy to operate,and have practical application value for the clinical detection of EDSV antibodies.