Preparation Of Goose Parvovirus VP3Monoclonal Antibody And The Study On ICS To Diagnose GPV

Goose parvovirus disease can caused a highly contagious and septic diseases in goslings and muscovy duck as mainly against the goslings that is4-20days old,which is caused by the Goose parvovirus (GPV). The disease spread fast, high morbidity and mortality. The mortality is up to100%.It,s serious harm to goose industry. Goose parvovirus is Parvoviridae, parvovirus,the nucleotide sequence contains two open reading frame (ORF), the right side of the ORF (RORF) encoding3structural proteins (VP):VP1, VP2, VP3. VP3is the major capsid protein and an important protective antigen of the virus, and can induce antibody with a neutralizing effect. Currently the disease is usually diagnosed by means of laboratory diagnosis, such as agar diffusion test, PCR test. However, the experimental method is more complex, time-consuming. So quick, simple and specific test method is more suitable for goose parvovirus in clinical diagnosis.Colloidal gold immunochromatography technology is simple, sensitive and specific, does not require special equipment, intuitively judge,etc. Colloidal gold immunochromatography technology is widely used in viruses, microorganisms, toxins and drug residues and other aspects of testing.The BALB/c mice were immuned by GPV-VP3prokaryotic expression protein, Preparation of monoclonal antibodies by hybridoma technology. The test obtained a hybridoma cell line(4E5)which can be secreting anti-GPV-VP3monoclonal antibody after repeatedly subcloning. Identified, the subtypes is IgGl, the light chain is κ chain. By western blotting analysis,the monoclonal antibody could specifically recognize GPV and VP3expression protein.The test purified monoclonal antibodies apply the method of octanoic acid-saturated ammonium sulfate, prepared colloidal gold particles by citrate sodium reduction,using colloidal gold particles labeled monoclonal antibody,respectively,rabbit anti-GPV IgG and goat anti-mouse IgG as the test line and control line on the NC membrane, Preparation of colloidal gold diagnostic test strip,and optimize the preparation conditions of the test strip.The results show that the combination optimal pH of monoclonal antibodies and colloidal gold is8.2,colloidal gold with the best combination of monoclonal antibody concentration is25μg/mL. Prepared under this condition,the colloidal gold test strip for the detection of goose parvovirus is the best. This study prepared anti-GPV-VP3monoclonal antibody,and prepared the colloidal gold diagnostic test strip with the monoclonal antibody.The colloidal gold diagnostic test strip is convenient for the goose parvovirus disease in clinical testing and a large number of testing in locations.It is laid the foundation for early detection of disease,early diagnosis,early treatment basis.