猪圆环病毒2型间接ELISA抗体检测方法的建立与初步应用

本研究利用原核表达的猪圆环病毒2型（PCV2）Cap蛋白作为标准抗原蛋白建立PCV2抗体检测方法，同时对建立的检测方法的相关条件进行了优化。通过优化确定抗原的最佳包被浓度为10 μg/mL，血清的最佳稀释度为1∶160，最佳封闭试剂为10%牛血清,二抗的使用浓度为1∶1000，血清及二抗的最佳反应条件为37 ℃孵育60 min，底物的最佳反应条件为37 ℃反应10 min，cutoff值为0.25。通过批内及批间重复性的评价，变异系数均小于0.1，表明本检测方法具有较高的可重复性。与标准的检测试剂盒相比，检测临床样本时，本检测方法与对照试剂盒检测结果符合率为91.7%。结果表明，建立的检测方法可用于临床PCV2抗体的检测。

Establishment of Antibody Indirect ELISA and its Initial Application

This study established an indirect antibody enzyme-linked assay system based on the expressed Cap protein. We optimized some factors such as antigen coating concentration (10 μg/mL), the dilution of serum samples (1∶160), blocking reagent (10% bovine serum), the work concentration of HRP-labeled anti-porcine IgG (1∶1000), reacted conditions of samples and HRP-labeled anti-porcine IgG (37 ℃, 60 min). The value of cutoff is 0.25. The CV is not beyond 0.1. In the detection of 60 clinical samples, the established ELISA showed excellent agreement with the compared kit (91.7%). The developed ELISA is suitable for PCV2 monitoring and detection, especially for screening large number of samples in farms.