Isolation And Identification Of Pseudorabies Virus Shaanxi WG Strain And Sequence Analysis Of GE And GD Gene

Pseudorabies virus (PRV), or named herpes virus I, is a member of Alphaher pesvirinae, herpesviridae, which can cause Aujeszky's disease in many domestic and wild animals. Its clinical symptoms are characterized in piglets by (PRV) fever, dyspnea, nerval symptom, salivation, growth stopping, weight falling, and high mortality, in sows by breeding disorder such as abortion and dead fetus, in grown-up animals by light symptom. PRV infection could cause its natural and main storing host-swine (also its main infectious origination) a huge loses, the most huge economical lose in all swine infectious diseases except Classic Swine Fever and Food and Mouth Disease. In order to discover the characteristic and epidemic rule and control the epidemic disease, we did some research on isolation and identification the WG strain of PRV, cloning and analysis of gE and gD gene.About 187 serum samples from 6 pig farms in some region of shannxi province were detected with the gE- ELISA Kit which can distinguish the vaccination from wild-type virus infection. The results showed that the average of positive rate was 28.76%,the highest gE-antibody of wild-type PRV was 48.98% and the lowest was 0, and that the PRV infection is very popular in shannxi province. Porcine pseudorabies virus was isolated from a field collected purtenance and the brain of a 2-day-old pig. To identify the virus microbiologically, chicken embryos were infected by field isolate and the typical pathology were observed. Infected PK-15 cell line shrunk, aggregated, and detached from the culture system in 20h approximately. When titrated in PK-15 cell culture, TCID50 attained 10-7.25/100μL, and it could be neutralized by the Min-A standard serum. Mock-infected rabbits were itching severely. We get the specific 612bp bands by PCR of the isolated virus.These data indicate that the isolated virus could be PRV and referred to as WG strain in this report.The isolates DNA were extracted and use to amplify gD and gE gene of PRV by olymerase chain reaction (PCR), 1203bp and 1734bp fragments were obtained .The PCR production were purified and cloned into pMD18-T vector. The recombinant plasmid were transformed into E.coli DH5α,then the positive clones were obtainded by Amp+ selection, identified by PCR and restriction digestion. The recombinant DNA were purified, and sequenced. The gD and gE gene sequences of PRV WG strain were compared with other PRV isolates in the Genbank, and nucleotide and amino acid were analyzed by use the software of DNAstar. Make phylogenic tree, reveal the genomic relatedness among different...