Abstract: The genome sequence of a duck hepatitis virus type 1 (DHV-1) strain was determined. Comparative sequence analysis showed that the genome has a typical picornarivus genetic organization, and strain DHV1-R genetic organaiztion is 5' UTR–VP0–VP3–VP1–2A1–2A2–2B–2C–3A–3B–3C–3D–3' UTR, DHV1-R has colsed realationship with Parechovirus, DHV1-R has 94.2%-99.2% nucleotide sequence homology and has98%-98.8% amino acid homology with other DHV1 strains ,Based on the DHV1 sequence database on Genbank,three pairs of specific primers were designed to amplify duck hepatitis virus type 1 using nested PCR and real-time PCR ,the results showed that sensitivity of nested PCR is 6pg/mL,and real-time PCR Tm value is 85.6℃ ,and the detected limit of postive plasmid is 0. 015fg/μL,all tests showed that nested PCR and real-time PCR have high sensitivity and specificity to detect duck haptitis virus type 1.