Study On The Analysis Methods Establishment And Application Of VP1 Gene Detection From Inactivated Vaccine Against Foot-and-Mouth Disease

Using Emulsion breaking purification antigen technology to deal inactivated vaccines against Foot- and-Mouth Disease, with alcohol, freezing &thawing and centrifuging.Extraction gene VP1 RNA from white oil emulsification with inactivated vaccines against FMDV, live virus and inactivated vaccine by BEI, RT-PCR amplified, cDNA sequencing and used SepMan of DNAStar Software package to connected together the sequences. The results indicate that, VP1 RNAs isolated from live virus, virus inactivated by BEI and inactivated vaccine emulsified by white oil access to all of the specific objectives of fragments, and the expected 639bp fragment match. Nucleotide sequence revealed that VP1 homology of the virus was 100% isolated by three methods. BEI and the emulsification process do not damage whole RNA structure of VP1. The isolated method to RNA can be used to analyse sequence of VP1 gene, which derived from either live or inactivated Foot-and-mouth Disease virus.Isolated VP1 gene by this methods to deal inactivated vaccine against foot-and-mouth disease, these inactivated vaccine come from three different manufacturers. The Sequence of VP1 gene between the gained sequence and other published sequence of FMDV compared and analyzed by the software of DNAStar, and Drew the phylogenetic tree. Analysis result showed that two manufacturers isolated FMDV belongs to PanAsia Genetic group, and the other is belongs to Cathay Genetic group. Between the 2 vaccine strain belongs to PanAsia, their homology of 96.2%, and compared them with recent international pandemic virus O/LY/2000, O/MOG/2000, O/1734/RUS/2000, O/NY00, UKG/8098/2001, the homologies among VP1 gene sequences are 94.4-98.4%.The experiment has built one kind of the pragmatic method that can extracted VP1 gene from the inactivated vaccine. To analysis of relationship between vaccine strains and epidemic strain, it's great practical importance to provided an effective and reliable vaccine for immune prevention of foot and mouth disease in China.