Evaluation of polyclonal IgG antibody in Response to injection of viral Hemorrhagic Septicemia killed virus(VHSv) of trout fish(Oncorhyncus mykiss) in rabbit(Oractulagus cuniculus) in order to produce rapid diagnostic kit

Aquaculture has become one of the sustainable sectors with a bright and promising future to ensure the safety of human food. Due to the need to increase the production of this healthy food, intensive aquaculture has been welcomed, which causes many diseases, especially viral. In recent years, VHS virus has caused irreparable damage to the country's trout fish producers. Rapid diagnosis and elimination of infected fish in the early stages of the disease is essential to control the spread of the disease effectively and reduce cost wastage, and rapid diagnostic kits will be very useful. These kits are made out of  antibodies. In this research, the production of polyclonal IgG antibodies against VHS virus was investigated in a New Zealand rabbit animal model(Oractulagus cuniculus). In order to this , the stock of  VHS GenBank Accession No: AF345859) Prepared by European Reference Laboratory as  an antigen to stimulate the immune system with Freund's complete adjuvant in four treatments including negative control of distilled water, treatment one with a concentration of 1 cc virus Complete, treatment two with a concentration of 0.5cc virus and 0.5cc adjuvant Freund, treatment three with a concentration of 75% virus and 25% adjuvant, treatment four with a concentration of 25% virus and 75% adjuvant, for four months, once a month IM In 5 groups of New Zealand rabbits injected, after 15 days, the ratio of blood sampling from the peripheral vein and antibody absorption (OD) were measured. During 4 blood samples, the amount of IgG antibody absorption was significantly different between the groups P <0.05) showed. Antibody purification was then activated by Sepharose Protein G adsorption chromatography column 4B (Sepharose® 4B) ) and OD of antibodies purified by Nano-200-Ver1.0 at 280 nm in the second treatment were 2.60 and the third treatment was 2.38 and the antibody concentrations were 1.82 and 1.67 mg, respectively. This antibody is going to be used in the rapid diagnosis kit of this disease.