抗羊口疮B2L囊膜蛋白单抗的免疫学初步研究

Preliminary Study of Immunology on Monconal Antibody Against Envelope Protein B2L of ORFV

探究一株抗羊口疮病毒B2L囊膜蛋白单抗的特异性识别毒株的能力,为后续利用单抗建立检测病毒的ELISA诊断试剂盒打下基础。通过将分离株OV/HLJ/04接种于牛睾丸细胞,并设立对照组,每天定时记录细胞病变过程。根据细胞病变情况选择最佳的免疫荧光时间,然后开展间接免疫荧光试验,并对产生病变的牛睾丸细胞进行PCR鉴定。病毒接种实验显示,细胞由接种病毒到产生病变的时间约3 d,3 d后接种细胞逐渐产生大规模的病变,实验选择2 d作为荧光实验的最佳时间。通过免疫荧光实验及PCR鉴定结果表明,所获单抗能够识别感染病毒的细胞。

In order to characterize capacity of identifying Off virus （OV） of the monoclonal antibody （McAb） against protein B2L, the protein B2L is the major envelope protein of ORFV.The research would lay the foundation of establishing the ELISA diagnostic kit of ORFV in the future.The isolated strains （named OV/HLJ/04） was inoculated at bovine testes cell （BT）,while non-inoculated cell was regarded as control group,and the dynamic changes of BT cells would be daily recorded in time.According to the choice of immunofluorescence on the optimal cell lesions,the indirect immunofluorescenee test was carried out,and the ORFV from the infected BT cells would be identified by PCR.The virus inoculation experiment showed that the time of the vaccinated cells against the virus to produce lesions was about 3 days,and the cells would largely produce CPE after 3 days,and 2 days had been chosen in the indirect immunofluorescence test as the best time.The experiment showed that MeAb could recognize the infected cells by immunofluoreseent antibody test and PCR.