Cloning Of The VEGF Gene Of Cantaginus Ecthyma Virus And Diagnosis Of CEV By PCR

The Orf-ys virus was propagated and the total DNA of virus was extracted.According to the published VEGF gene sequence of ORF V strain in GenBank, one pair of primer was designed. The VEGF gene of virus was amplificated with the primer by PCRand then cloned into pMD18-T plasmids and then sequenced and analysed in homology.The acquired sequences contained 414bp, The homology analysis revealed that thehomology of the strain Matsumoto and the strain Orf-ys was 98%, the IT01 and the Orf-ys was 84%. There were a little variation between Orf-ys strain and the referenced virus strains.According to the published sequence of strain NZ2 in GenBank, another pair of primer was designed. The virus DNA was amplificated with the primer by PCR, then PCR diagnostic method was established, and this method's specificity and sensitivity was detected. The results indicated that the PCR method could detect the nucleic acid of Orf-ys virus only and a 414bp DNA fragment was amplificated. The minimum ORF V DNA's detection level was 4.5ng. This illustrated that this method was specific and sensitive for the detection of ORF V.