兔病毒性出血症病毒间接ELISA抗体检测试剂盒的研制及其临床应用

Development of the Detection Kit and Its Clinical Application of an Indirect ELISA for Detecting Antibodies to Rabbit Hemorrhagic Disease Virus

为了建立一种快速的兔病毒性出血症病毒抗体检测方法,本研究参照已发表的RHDV基因序列,RT-PCR扩增了长约510bp的VP60基因片段,连接PGEX-4T-1表达载体后获得了以包涵体形式表达的重组VP60蛋白。重组蛋白纯化后,经免疫印迹检测证明具有良好的抗原性和特异性。以该蛋白作为诊断抗原,建立了检测兔病毒性出血症病毒抗体的VP60-ELISA诊断方法。该诊断方法具有良好的敏感性、特异性和重复性,为RHDV的快速诊断、免疫兔群抗体监测和实验兔等级检测提供了一种快速、简便的血清学诊断方法。

In order to establish a rapid rabbit haemorrhagic disease virus antibody detection method,this study amplificated a length of 510bp VP60 gene fragment refered to the published RHDV gene sequences.The purified recombinant VP60 protein as coating antigen was obtained after the target fragment was directionally cloned into expression vector PGEX-4T-1.As reparation of recombinant VP60 protein,this recombinant protein has a good antigenicity and specificity.This experiment established the VP60-ELISA diagnosis method to detect the rabbit haemorrhagic disease virus antibody.This method has good sensitivity,specificity and reproducibility and could be used as a simple and rapid serology detection method for monitoring of anti-RHDV antibody and epidemiologic survey of RHDV.