牛病毒性腹泻病毒1型灭活疫苗的制备及其免疫效果评价

Preparation and immune evaluation of bovine viral diarrhea virus type 1 inactivated vaccine

为研究针对牛病毒性腹泻病毒1型(BVDV1)毒株的灭活疫苗,选用本实验室分离的分离株SMU-Z6/1a/SC/2016和SMU-DJ2/1d/QH/2015与201佐剂制备BVDV水包油包水灭活疫苗免疫BALB/c小鼠或奶牛,并通过抗体中和试验、抗体交叉保护试验及ELISA方法对免疫后的抗体消长规律进行研究。结果显示,分离株SMU-Z6/1a/SC/2016制备的灭活疫苗免疫小鼠后产生的中和抗体相比分离株SMU-DJ2/1d/QH/2015制备的灭活疫苗高,最高值为1∶177;抗体交叉保护试验表明,SMU-Z6/1a/SC/2016灭活疫苗免疫机体产生的抗体可以保护分离株SMU-DJ2/1d/QH/2015;而SMU-Z6/1a/SC/2016灭活疫苗免疫奶牛后抗体效价最高值为1∶1 413,且高于商品化疫苗组。基于分离株SMU-Z6/1a/SC/2016 E2重组蛋白建立的ELISA检测结果表明,SMU-Z6/1a/SC/2016灭活疫苗也能刺激机体产生较高水平的抗体,且整个试验过程中均高于商品化疫苗组,该结果与抗体中和试验结果一致。结果表明,以分离株SMU-Z6/1a/SC/2016制备的灭活疫苗可刺激不同动物机体产生高水平的中和抗体,这为我国BVDV灭活疫苗的研制和疫苗候选株的筛选提供了一定的数据支持和指导意义。

This study aims to explore the inactivated vaccine with bovine viral diarrhea virus type1(BVDV1) strain.In this study,two strains SMU-Z6/1 a/SC/2016 and SMU-DJ2/1 d/QH/2015 were isolated from yak feces and preserved in our laboratory.The 201 adjuvant was used to prepare BVDV inactivated vaccine to immunize BALB/c mice or cows.After immunizing,the antibody titers were detected by neutralization test and ELISA methods.In result,the neutralizing antibodies in mice with the SMU-Z6/1 a/SC/2016 vaccine have higher titers compared to the SMU-DJ2/1 d/QH/2015 vaccine,with the maximum of 1 ∶ 177,The antibody cross-protection experiments showed that the antibody with the SMU-Z6/1 a/SC/2016 vaccine could protect the SMU-DJ2/1 d/QH/2015 strain.Furthermore,the highest antibody titer with the SMU-Z6/1 a/SC/2016 vaccine was 1 ∶ 1 413 in cows,which was significant higher than that of commercial vaccine group.The ELISA based on recombinant E2 protein was developed to detect the BVDV antibody in cow.The SMU-Z6/1 a/SC/2016 inactivated vaccine could produce higher antibodies levels than those of the commercial vaccine,which was agreed with the results of the antibody neutralization test.The inactivated vaccine prepared by SMU-Z6/1 a/SC/2016 isolate could stimulate the relative high antibodies in mice and cows.Therefore,our study provides some data to develop the BVDV inactivated vaccine and to screen the vaccine candidate strains in China.