Preparation and application of VP3 antiserum of infectious pancreatic necrosis virus
CHEN Guihua, HE Wenbin, XU Liming, ZHAO Jingzhuang, REN Guangming, SHAO Yizhi, DUAN Kaiyue, LU Tongyan
1.National Demonstration Centre for Experimental Fisheries Science Education, Shanghai Ocean University, Shanghai 201306, China; 2.Heilongjiang River Fisheries Research Institute, Chinese Academy of Fishery Sciences, Harbin 150070, China; 3.Aquatic Pathogen Collection Center of Ministry of Agriculture and Rural Affairs, Shanghai Ocean University, Shanghai 201306, China; 4.Yangtze River Fisheries Research Institute, Chinese Academy of Fishery Sciences, Wuhan 430223, China
Abstract: In order to enrich the detection methods of infectious pancreatic necrosis virus(IPNV), the gene sequence(711 bp)encoding VP3 gene of IPNV was amplified by one-step RT-PCR using genomic RNA of Genogroup Ⅰ IPNV strain ChRtm213 as template.The VP3 protein was expressed in Escherichiacoli rosetta, and the purified VP3 protein was used as antigen protein to prepare mouse antiserum.The titer of antiserum was determined by enzyme linked immunosorbent assay(ELISA), and the ability of antiserum to recognize the current IPNV isolates in China was identified by indirect immunofluorescence antibody test(IFAT).The SDS-PAGE analysis showed that the purified VP3 protein was a single band with a relative molecular mass of 45 000, which was consistent with the theoretical size of the VP3 protein.The titer analysis of antiserum showed that the titer of the prepared anti IPNV VP3 serum was 16 000 in IPNV and 32 000 in purified IPNV VP3 protein.IFAT results showed that the prepared mouse antiserum specifically recognized Genogroup Ⅰ and Genogroup Ⅴ IPNV isolates which were isolated from different regions in China.Moreover, the VP3 mouse antiserum did not cross react with infectious hematopoietic necrosis virus(IHNV)and viral hemorrhagic septicemia virus(VHSV), showing high specificity with IPNV.These findings indicated that the VP3 protein expressed in the prokaryotic expression system had promising immunogenicity, and the prepared IPNV VP3 mouse antiserum specifically recognized IPNV isolates from different regions in China, which laid a foundation for the immunological detection of IPNV in China.
2021, Vol. 36 
