Development And Application Of Monoclonal Antibodies Against Canine Parvovirus

Canine parvovirus (CPV-2) is a significant pathogen for domestic dogs. It causes an acute haemorrhagic diarrhoea and sometimes fatal myocarditis in young dogs.CPV affects puppies much more frequently than it affects adult dogs. In very young puppies it can infect the heart muscle and lead to "sudden" death. This newer virus seems to represent a mutation from the feline parvovirus (which is more commonly known as the "feline distemper virus"). Because this virus was (and is) shed in gigantic numbers by infected animals and because this virus is especially hardy in the environment, worldwide distribution of the virus rapidly occurred.Four from ten clinical samples were dectected positive with CPV colloidal gold test paper. By Fk81 cell culture, as a result, one CPV strains (No.2) were isolated successfully. By using the CPV as antigen, Monoclonal antibody (McAb) was prepared, and two positive clones, designated as 4E4 and 5C8 respectively, were obtained from hybridoma cell lines.The BALB/c mice were immunized subcutaneously with CPV, which was purified with polyethylene glycol. After being immunized three times, the mice were immunized intraveinally with equal the dose for the final immunization. Three days later, spleen cells from immunized mice were fused with SP2/0-Ag-14 myeloma cells. Indirect enzyme linked immunosorbent assay (iELISA) was used to screen hybridoma cells, and limiting dilution method was applied to subclone positive hybridoma cells three times, and two positive clones, designated as 4E4 and 5C8 respectively, were obtained from hybridoma cell lines. 4E4 was subtyped to be IgM, and 5C8 was subtyped to be IgG.1 .Two hybridoma cell lines secreting McAb against CPV have been established with the characteristic of counteraction in four places, such as Yangzhou. And they were applied to treating CPV-infected dogs, the effect was obvious.