牛肠道病毒3D抗体与感染相关性研究

Study on the relationship between the antibodies level of Bovine enterovirus 3D protein and virus infection

为了研究感染牛肠道病毒后非结构蛋白3D的抗体消长规律,试验从牛肠道病毒HLJ-3531株中扩增3D基因,将其克隆至表达载体pET-30a并在大肠杆菌Rosetta(DE3)中表达,以Ni-NTA柱亲和层析法纯化得到的重组3D蛋白为抗原制备3D多克隆抗体,随后建立3D-ELISA方法用于检测HLJ-3531毒株感染小鼠后的3D抗体消长规律,同时用RT-PCR方法检测感染时期小鼠体内病毒核酸。结果表明:RT-PCR技术可检测出病毒出现的时间范围与3D抗体存在的时间范围符合性良好。说明3D-ELISA配合核酸检测方法有很高的概率检测出病毒感染活动期动物。

The aim of the present study was to detect the growth and decline rules of non-structural protein 3D antibodies after animals infected by Bovine enterovirus. The 3D gene was amplified from the genome of Bovine enterovirus HLJ-3531 strain,then cloned into the expression vector pET-30 a and expressed in Escherichia coli Rosetta( DE3). The recombinant 3D protein purified by Ni-NTA column affinity chromatography was used as antigen to prepare 3D polyclonal antibody. Then a 3D-ELISA method was established to detect the growth and decline rules of 3D antibody in mice infected with HLJ-3531 strain. Meanwhile,the RT-PCR method was used to detect the viral nucleic acid in the infected mice. The results showed that the existence time of the virus detected by RT-PCR was consistent with the existence time of the 3D antibody. Accordingly,there was a high probability that the animals infected with the active viruses could be detected by 3D-ELISA and nucleic acid detection methods.