Isolation And Identification Of Porcine Epidemic Diahorrea Virus Stain Bd And Establishment Of Vero Cell Line Stable Expressing Porcine Aminopeptidase N Gene

Porcine epidemic diarrhea virus （PEDV） causes lethal diarrhea in piglets that leadsto greateconomic losses in all word. It belongs to group I coronavirus.Porcine epidemic diarrhea virus（PEDV）strain named BD was isolated from pigletssuffering from a severe diarrhea in BaoDing city using Vero cell. The experimental resultsindicated that the bland passages of22genrations were undertaken untill the CPE could beseen on23genrations. The isolated virus was identified by stable cytopathic effects（CPE）, RT-PCR,IPMA, ELISA,TCID50ect. The experimental results indicated that thebland passages of22genrations were undertaken untill the CPE could be seen on23genrations in48h. PEDVs were identified through RT-PCR on the M gene in order tocertificated that the CPE caused by PEDV and the viruspropagated in Vero cells. Theresults were masculine. Immunoperoxidase monolayer assay（IPMA） test indicated thatviral antigen could be detected and could be seen brown in Vero cells. Indirect ELISAdetected that OD450nm≥0.260.It was justified that the virus in Vero cell culture wasmasculine. Titer of the virus was10-2.5/0.05ml at23passage.Above experiments certifiedthat it was PEDV from the feces and it propagated in Vero cell culture in mediumcontaining trypsin. it can provide materials for serology immunology and biologicalcharacteristics of PED, and provide a theoretical basis for the effective prevention andcontrol of PED.