Cloning, Sequence Analysis And Studies Of Nucleic Acid Probe Of The F1L Gene Of Contagious Ecthyma Virus

A Vstrain(OV-nm-05)Contagious Ecthyma Virus (CEV)isolated in Inner Mongolia was propagated,and the total DNA of virus was extracted.According to the published F1L gene sequence of NZ2 strain in GeneBank,two pairs of primer were designed,one was sequencing primer,the other was diagnostic primer.The F1L gene of virus was amplificated with sequencing primer by PCR and then cloned into pGEM-T Easy plasmids and then sequenced.The acquired sequences contains 1005bp,and congtain one ORF and the nontranslated region.The acquired sequences of OV/nm-05 was compared with OV/7, NZ2, OV/C2, OV/mi-90, OV/Torino and OV/20.The results showed OV-nm-05 had the hingest homology (99.1%)with OV-mi-90. The homology anaysis revealed that there were a little variation between OV/nm-05 strain and the referenced reovirus strains.Using the technique of dot blot hybridzatiom,the DNA of CEV was detected with the F1L gene labeled with Dig-dUTP .Detection of sensitivity and specificity showed the dot blot hybridzatiom had hing sensitivity and specificity.It reacted positively with DNA of CEV only and did not reacted positively with DNA of NDV,IBDV,IBV.This kind of probe could even detect 20pg/s.