Evaluation on the Efficacy and Immunogenicity of Recombinant DNA Plasmids Expressing Spike Genes from Porcine Transmissible Gastroenteritis Virus and Porcine Epidemic Diarrhea Virus

Meng, Fandan; Ren, Yudong; Suo, Siqingaowa; Sun, Xuejiao; Li, Xunliang; Li, Pengchong; Yang, Wei; Li, Guangxing; Li, Lu; Schwegmann-Wessels, Christel; Herrler, Georg; Ren, Xiaofeng

Serum IFN-c levels were analyzed with an IFN-c detection kit (Excell Bio., China) according to the manufacturer's instructions. A standard curve was generated using control IFN-c diluted in PBS at different concentrations beginning with 10,000 pg/ml followed by two-fold serial dilutions between 2000 pg/ml and 61.25 pg/ml. Dilutions were subsequently coated onto ELISA plates overnight at 37uC. Sera (1:100) from 42 dpi mice were also coated onto ELISA plates and used as primary antibodies in a parallel experiment to evaluate the virus-derived IFN-c response. HRP-conjugated goat-anti mouse (1:2000) was used as secondary antibody in both the control experiment and in the analysis of the mouse sera. The OD 490 values and therefore pg/ml of IFN-c in immunized mice were determined relative to the IFN-c standard curve.