Study On Biological Properties And PCR Detecting Method Of Contagious Ecthyma Virus

From the scab of the sick sheep ,one strain of virus was isolated using Bovine Testicle cell and MDBK cell,then,it was not taken cell passaged until the CPE is order .the recurrent infection,replication test, Aether and Chloroform-stability test, other biological property, and electrongraph test proved it was Contagious Ecthyma Virus(CEV)According to the published sequence of F1L gene of the CEVin GeneBank, a pair of differential primers was designed by oligo 4.1. The primers are PB1 : CAC GGG CTA CAT AAT CGG GGT TGC PB2 : TTG CGG AAG GTC ATG TCG TTG TCG. Wegot the optimal condition was got for PCR systemic by optimizing and filtrating. The optimal denaturing temperature was 94℃, the optimal anneal temperature was 55℃, the optimal circles were 35 times. At the same time, the premium result of the PCR system was also found by experimentation.A 429 bp specific fragment was amplified in PCR experiments with DNA from the Infectious CEV, the amplified fragment accorded with the predicted amplification fragment. However, the PCR did not amplify the specific fragment with DNA from FMDV,MDBK cell.The results showed that this PCR amplification system was specific.In addition, the PCR system could detected 0.5 ng dilute template. It showed that The sensitivity of the PCR was very high. According to the above results, It was showed that the PCR detecting method was to be specific and sensitive.