Development Of Colloidal Gold Immuno Strip For The Detection Of Chinese Sacbrood Virus

Sacbrood which caused by Sacbrood virus(SBV) is a severe infectious disease in honeybees, especially in the Apis cerana called Chinese Sacbrood virus(CSBV). It can induce an enormously high mortality rate in larvae and also affects on the foraging and breeding activities of adult bees. However, there is no effective detection technique that can be directly applied in apiculture.In this study, the CSBV from Jiangxi province was used as experimental material.Nucleotide and protein sequences of vp1 and vp3 gene of CSBV were obtained by clone and then were analyzed with phylogenetic analysis. After undertaking the prokaryotic expression of vp3, the monoclonal antibody of CSBV was created. Furthermore, peptides antigen was designed based on the vp1 protein sequences of CSBV and was used as antigen to produce polyclonal antibody of CSBV. Finally, Based on monoclonal antibody and polyclonal antibody, we preliminary developed a colloidal gold test paper strip of CSBV by adopting the colloidal gold immuno chromatography combining with double antibody sandwich method. The main results are as follows:1. The nucleotide sequence of vp1 gene of this CSBV was 973 bp which may encode324 amino acids. The nucleotide sequence of vp3 gene was 1070 bp, encoding 356 amino acids.2. Sequence analysis of vp1 and vp3 genes of CSBV showed that the similarity of vp1 gene among Jiangxi CSBV and other isolates was lower than that in vp3 gene,suggesting that vp3 gene is more conservative compared to vp1. Based on the analysis of SBV complete genome, vp1 gene and its conservative fragment, we therefore speculated that the host specificity and the geographical distribution of SBV were both determined by their genetic diversity, and there might be a cross-infection of SBV between Apis. cerana and Apis. Mellifera.3. The monoclonal antibody and polyclonal antibody of Jiangxi CSBV were developed by which a CSBV colloidal gold test strip was preliminarily designed. Our results showed that the SBV could be detected by CSBV colloidal gold test strip.