Abstract: To identify suspected goats Orf case and analyze the molecular characteristics of Orf Virus (ORFV), crust materials of lambs with clinical sore mouth symptom from a farm in HuBei Province were used and researched. ORFV in tissue scrapings from the lips was confirmed by electron microscope, HE stain and specific gene(B2L)PCR amplification. Molecular characterization of ORFV strain was performed using amino acid sequence homology comparison and phylogenetic analysis based on B2L gene. The results showed that this disease was caused by ORFV(named TS/HuB/CHIA/2009), B2L gene sequence was submitted to GenBank and obtained accession number GU320351. Phylogenetic tree analysis demonstrated that the GU320351 has a closest genetic relationship with DQ904351, which was isolated from Taiwan Province of China in 2007. And GU320351 was in the evolutionary branch contain DQ263306, DQ263305, DQ263304 and DQ263303, which was isolated from India in 2004 and 2005. Research of this paper accumulated useful material for further studying of molecular epidemiology of ORFV in China.