Research On The Rapid Determination Of The Antigens Of Porcine Parvovirus

Porcine parvovirus infection, which is caused by Porcine parvovirus, a DNA virus belonging to the family Parvoviridae, genus Parvovirus, is characterized by reproductive failure in sows. And it can cause persistent infection among swine herds with the infection rate reaching up to 90%. Clinically it is hard for serological tests to detect the infection due to the fact that PPV is characterized by being a long-term carriers of its virulence, and persistent infections with low-dose of PPV frequently occur. In addition, laboratory tests are not only restricted by specific apparatuses and environment, but also time-consuming. To avoid these detection defects above, lateral-flow immunochromatographic test strips featuring the properties of high specificity and simplicity were produced in this study.Since the low titer and poor immunohistochemistry results of the commercial HRP conjugated antibodies, IgG extracted from pigs and rabbits was purified to immunized goats, which in turn resulted in the production of high-titer antibodies(Antibody titer:1:160-320). Those antibodies were conjugated with HRP by SATP-SMCC methods. Thus, goat anti pig IgG-HRP conjugation with the antibody titer of 1:64000 was obtained.Apparent cytopathic effect(CPE) was observed upon PPVs were propagated passaged five times in PK-15 cell line cultured by DMEM. And polyclonal antibodies with the titer of 1:51200 were obtained after immunizaion of rabbits by the purified virus.After amplification and purification of PPV, it was used to immunize BALB/c mice. PPV mAb was produced through cell fusion and the screening of specific hybridoma cell lines. Thus, the test strip for the rapid detection of PPV were developed with colloidal gold -labeled (MACG) technique, and it could detect virus fluid quantitatively diluted 640 times. What's more, the test strip shows no cross-reactions with CSFV, JEV and PRRSV. Therefore, characterized by rapid detection, high sensitivity, specificity and simplicity, the test strip is capable of being widely-used in clinics and productions.