论文部分内容阅读
为建立一种检测水貂犬瘟热病毒(CDV)血清抗体的间接ELISA方法,将CDV的血凝素蛋白(H)基因的主要抗原表位片段克隆至p ET-30a-c(+)载体中,转化至感受态细胞Rosetta(DE3)中进行诱导表达,并对表达产物进行SDS-PAGE分析和Western-blot鉴定。以纯化的重组H蛋白作为包被抗原,建立了检测CDV抗体的间接ELISA方法,并证实该方法具有良好的特异性、重复性和敏感性。采用建立的方法与国外商品化的犬瘟热抗体检测试剂盒对80份血清进行检测,两者符合率为92.5%。本研究建立的间接ELISA方法为水貂抗体水平检测和评价犬瘟热疫苗免疫效力提供了简便快速的方法。
To establish an indirect ELISA for the detection of serum antibodies to mongrel canine distemper virus (CDV), the major epitope fragment of the hemagglutinin (H) gene of CDV was cloned into the pET-30a-c (+) vector , Transformed into competent cells Rosetta (DE3) induced expression, and the expression product was analyzed by SDS-PAGE and Western-blot identification. The purified recombinant H protein was used as coating antigen, and an indirect ELISA method for detecting CDV antibody was established, and the method was proved to have good specificity, repeatability and sensitivity. Eighty sera were tested by the established method and commercialized Canine distemper antibody test kit, the coincidence rate was 92.5%. The indirect ELISA method established in this study provided a simple and rapid method for the detection of mink antibody level and evaluation of the immunogenicity of the distemper vaccine.