Identification Of The Xinjiang 's Isolate Of Pseudorabies Virus And Sequence Analysis Of The GD And GE Gene

One Pseudorabies( Aujeszky's disease) virus isolated from infected domestic pigs originating from Xinjiang.The　organ samples were filtered then drained monolayers of BHK-21 cells,the typical cytopathic effect(cpe) were observed. The virus appeared typical Pseudorabies virus form.The virus were neutralized with standard positive porcine sera using serum neutralization test(SNT). TCID50 of virus was 10-10.87/0.1mL. Rabbits and pigs showed typical syndrome of PR when they were inoculated with cells virus. PCR technique was applied for the detection PRV in cells virus and Amplified a fragment with desired size of 217bp.According with virus characterisation from infected pigs,the virus was indentified as PRV and named XIN-W strain。 To study the glycoprotein gene characteristics and molecular epidemiological background,PRV gD and gE gene sequence of XIN-W strain were determined. According to the published PRV gD and gE gene nucleotide sequence,two pairs of specific primers designed and synthesized. A 1209bp gD and a 1743bp gE gene completed sequence were amplified by polymerase chain reaction (PCR) method and colned into pGEM-T-Easy vector. After transforming the recombinant plaxmid to JM109,positive colnes were selected then confirmed by PCR and restrict enzyme identification.After the gD and gE gene nucleotide sequencing,the sequences of nucleotide and predicted amino acid were compared with those of other published PRV strains.The results showed that gD and gE gene nucleotide sequence homology respectively is 97.9%-99.3%,98.0%-98.7%; deduced amino acid sequence homology respectively is 97.0%-99.3%,97.1%-98.1%.At the nucleotide and amino acid level, gD and gE gene sequences were respectively compared with others published PRV strains, their homology were very high. Molecular phylogenetic tree show that XIN-W and others PRV strains isolated in China might originate from same source and belong to same evolution embranchment. By the pathogeny and molecular level, the study gives a preliminary proof that wild virulent strain of PRV exited in Xinjiang region. The research also has a great significance in exploration of molecular epidemiology of PRV and the development of diagnosis and prophylaxis against PRV.