Nested PCR detection and sequence analysis on Zhejiang duck circovirus

Using the nested PCR technology,the duck circovirus(DuCV) nucleotide fragment,which was isolated from Wenzhou,Zhejiang,was amplified and its sequence analyzed.The result showed that the obtained segment was about 340nt in length.Based on the determined sequence,two primers were applied for the construction of the remaining genomic sequence by inverse PCR.The genome of the Zhejiang DuCV had a length of 1995nt.Its ORFV1 encoded a replication protein of 292 amino acids,while the ORF-C1 encoded a capsid protein of 257 amino acids.The strain had highest sequence similarities(i.e.,99.2%) with EF451157 detected from the muscovy duck and EU344804 detected from the mule duck in Fujian.Phylogenetic analysis with Lasergene v 7.1 DNAStar software by Clustal W method showed that it had the same sub-lineage with the European strain detected in German mulard duck and the North American strain found in the American Perkin duck.