猫杯状病毒抗体间接ELISA检测方法的建立

Development of indirect ELISA for detection of antibodies against Feline Calicivirus

将构建的猫杯状病毒(Feline Calicivirus,FCV)衣壳蛋白VP1重组表达质粒转化感受态细胞,以表达的FCV-VP1蛋白作为包被抗原,以酶标记的兔抗猫IgG作为二抗,建立检测猫杯状病毒抗体的间接ELISA检测方法。确定了最佳封闭液、最佳血清稀释液、最佳酶标二抗稀释液,当酶标记的兔抗猫IgG效价为1∶20 000时可以检出猫杯状病毒血清抗体。利用本实验室建立的检测方法对收集的96份临床血清进行检测,结果表明96份猫血清中检测出40份阳性。

The gene of capsid protein VP1 from Feline Calicivirus(FCV)was cloned and expressed in Escherichia coli.Using the purified VP1 protein as coating antigen and Rabbit anti-cat IgG linked with HRP as secondary antibody,an indirect enzyme-linked immunosorbent assay(ELISA)for detecting antibodies against FCV was developed.The reaction condition was optimized.A total of 96 clinical samples of cat serum were tested and 40 serum samples were positive for antibodies against FCV.