应用HEV ORF2蛋白McAb间接免疫荧光法检测HEV的研究

应用抗猪戊型肝炎病毒（HEV）ORF2蛋白单克隆抗体和猪戊型肝炎病毒多克隆抗血清对感染猪戊型肝炎病毒的A549细胞培养物进行了间接免疫荧光检测病原的比较研究。单克隆抗体（McAb）进行的间接免疫荧光染色后,HEV感染细胞培养物中多数细胞在其胞浆和细胞膜上出现黄绿色荧光,未接种病毒的细胞培养物被伊文思蓝染成红色,未见有黄绿色荧光染色的细胞存在;多克隆抗血清对接种病毒的细胞培养物染色后,荧光强度高,呈现亮绿色,但未接种HEV的细胞培养物亦有一定的非特异性荧光反应。结果表明,利用所研制的抗HEVORF2蛋白McAb进行间接免疫荧光法对病原检测具有很高的特异性。

Detection of HEV with Indirect Immunofluorescent Assay by Monoclonal Antibody Against ORF2 Protein of HEV

The A549 cell culture infected with HEV was carried out for comparative studies on detection of pathogene with indirect immunofluorescence by using monoclonal antibody against HEV ORF2 protein and polyclonal antisera against HEV.The result indicated that the majority of cells in A549 cell culture infected with HEV showed strong green fluorescence in cytoplasm and cell membrance,the cell culture not inoculated with HEV was stained red by Evance blue,and there were no strong green fluorescent cells after indirect immunofluorescent assay by McAb.The cell cultures which were inoculated or not inoculated with HEV were detected using indirect immunofluorescenct assay by polyclonal antisera against HEV.The result indicated that the cell culture inoculated with HEV showed strong green fluorescence.But the cell culture not inoculated with HEV existed certain non-specific fluorescent response.The McAb against HEV ORF2 protein used for detection of pathogene with indirect immunofluorescence by developed ORF2 protein against HEV has strong specificity.