| Duck viral hepatitis(Duck viral hepatitis,DVH)is a highly infectious disease caused by Duck hepatitis virus(DHV),which has a short incubation period and a high mortality rate.It is an important disease of economic loss for duck production.Duck viral enteritis(Duck plague,DP)is an acute septic infectious disease caused by duck enteritis virus(DEV),which is characterized by digestion Organ damage and degeneration of solid organs.In 1957,China's initial outbreak of the disease,followed by nation-wide range of popular duck caused serious economic losses.In this study,the VP0 primers based on the sequence of DHAV-1 sequence were designed,and VP0 gene was amplified by polymerase chain reaction(PCR).The pET30a-VP0 vector was successfully constructed,and the VP0 protein was expressed and purified.The purified protein was used to immune New Zealand large rabbit by injecting abdominal.The immune titer was measured after three times of immunization.The rabbit polyclonal antibody against VP0 protein was successfully prepared.Using the system for successfully constructing DEV by transfection fosmids.The pDONR-PCA-VP0 was generated by BP reaction with pCAGGS-VP0.Then PCA-VP0 expression cassette was introduced into 2 fosmid by LR reaction,generating 2-UL41-VP0.The recombinant virus rDEV-VP0 was constructed by using 2-UL41-VP0 replacing 2 fosmid.The rDEV-VP0 was passaged in chicken embryo fibroblasts,then the inserted VP0 was identified by PCR.The result indicated that we successfully constructed the recombinant virus rDEV-VP0.To investigate the protective efficacy of the rDEV-VP0,we divided 60 ducklings into 2 rDEV-VP0 immunization groups,C-KCE immunization group,DEV challenge control group,DHV challenge control group,and control group.The DEV and DHV were challenged at 4 days post immunization.The results showed that rDEV-VP0 can provide full protection against lethal Chinese standard challenge(CSC)DEV and conferred 70% protection against DHAV-1 161/79. |
PDF Full Text Request