传染性法氏囊病(infectious bursal disease virus,IBD)是由传染性法氏囊病病毒(infectious bursal disease virus,IBDV)引起的一种主要危害雏鸡的急性传染病.在IBDV编码的 5 个蛋白中,VP5 是IBDV唯一可以缺失基因的蛋白.本研究克隆了IBDV优势流行毒株的VP5 蛋白编码基因,利用原核表达系统表达了VP5 蛋白,通过免疫BALB/c小鼠制备VP5 多抗,并对多抗进行了鉴定和检测不同IBDV毒株的应用.结果显示,IBDV优势流行毒株的VP5 蛋白实现了可溶性表达,分子质量约为 17 kDa;制备的VP5 多抗特异性良好,ELISA效价可达 1:64 000;VP5 多抗可通过IFA和Western Blot检测识别IBDV rHLJ0504-HT株,不识别VP5基因缺失株.结果表明:试验成功表达了IBDV优势流行毒株的VP5 蛋白,并制备了VP5 多抗;VP5 多抗可对IBDV及其VP5 编码基因缺失毒株进行鉴别检测.本研究对于IBDV检测方法的建立以及VP5 编码基因功能的进一步研究提供了物质基础.
Infectious bursal disease(IBD)is an acute infectious disease caused by infectious bursal disease virus(IBDV),mainly posing a threat to chicks.For 5 proteins encoded by IBDV genome,VP5 is the only one of which the gene may be deleted.In the study,the gene encoding VP5 protein(VP5 gene)of the dominant strain of IBDV was cloned,VP5 protein was expressed by prokaryotic expression system,and VP5 polyclonal antibodies were prepared through vaccinating BALB/c mice,then its application in various IBDV strains was identified and tested.The results showed that VP5 protein of IBDV dominant strain was expressed with solubility,and its molecular weight was about 17 kDa;the prepared VP5 polyclonal antibodies were with good specificity,and the ELISA titer could reach 1:64 000;VP5 polyclonal antibodies could identify IBDV rHLJ0504-HT strain through IFA and Western Blot,but fail for VP5 gene-deleted strain.In conclusion,VP5 protein of the dominant strain of IBDV was successfully expressed,and VP5 polyclonal antibodies were prepared,which could be used to identify and test IBDV wild and VP5 gene-deleted strain.A material basis was provided by the study for the development of IBDV detection methods and further research on the function of VP5 gene.