Isolation and Identification of Iridovirus and Preparation of Its Polyclonal Antibody in Diseased Large Yellow Croaker
A virus was isolated and identified in diseased large yellow croaker out broken in a fish farm in Xiangshan,Zhejiang Province,in July 2021 by epithlioma papulosum cyprinid(EPC)culture,and major capsid protein(MCP)cloning and sequencing.The tissue homogenate of diseased fish caused typical cytopathic effect including plaque,exfoliation in EPC cells.The amplification of conserved region of ATPase and major capsid protein(MCP)gene of iridovirus by PCR showed that 740 bp and 1 367 bp specific genes were obtained.Sequence MCP gene amplified fragment,and phylogenetic analysis revealed that the isolated virus belonged to the genus Megalocytivirus in Family Iridovirade.The purification of the virus by sucrose density gradient centrifugation and transmission electron microscopy observation found that the virus particles were regular hexagon in shape with diameter of 120-150 nm.Polyclonal antibody against large yellow croaker iridovirus was obtained by immunizing mice with purified virus as antigen,with the titer of 1∶7 000.Three immune proteins were preliminarily identified by SDS-PAGE and Western blotting.The finding provided a method for the purification of large yellow croaker iridovirus,identification of the immune protein,and theoretical basis for molecular biology research,protein research and vaccine preparation.
large yellow croakerIridoviridaeMegalocytivirusisolation and identificationvirus purificationpolyclonal antibody
万方数据
维普
