Molecular Characterization and SYBR Green I-Based Quantitative PCR for Duck Hepatitis Virus Type 1

摘要: Abstract To determine the genomic sequence of a duck hepatitis virus type 1 (DHV-1) strain, real-time quantitative polymerase chain reaction (RTQ-PCR) assay based on SYBR Green I technology was developed to target 3D gene DHV-1. Comparative analysis showed that genome has typical picornarivus genetic organization, and strain DHV-1 R organaiztion is 5′ untranslated region (UTR)-VP0-VP3-VP1-2A1-2A2-2B-2C-3A-3B-3C-3D-3′ UTR, close relationship with Parechovirus, 95.1-99.1% nucleotide identity other strains. Based sequences in GenBank, three pairs specific primers were designed amplify using PCR. The results PCR Tm value 85.6°C provides broad dynamic range, detecting from 102 109 copies cDNA per reaction. No cross-reactions found specimens containing DPV, AIV NDV. It concluded belongs new group family Picornaviridae may form separate genus most closely related Parechovirus. All high sensitivity specificity detect dissociation curve analysis, isolates can be distinguished by their melting temperature. These methods are rapid, sensitive, reliable, readily adapted for detection clinical samples.