Abstract:To develop a RT-PCR assay for the detection of novel duck reovirus(NDRV) and apply the developed assay to isolated strains and artificial infected samples, according to NDRV-NP03 S3 gene in the GenBank(GenBank:GQ888710), a pair of primers for amplying NDRV specifical fragment were designed and synthesized.RT-PCR technique detecting the RNA of NDRV was constructed.RT-PCR results showed that a 586 bp specifical fragment could be isolated only from the NDRV-NP03 strain RNA, and the sensitivity of RT-PCR reached to 2 pg NDRV-RNA.And the negative results were achieved from the other viruses, Muscovy duck reovirus(MDRV), Avian reovirus(ARV), Infectious bursal disease virus(IBDV), Muscovy duck parvovirus(MDPV), Goose parvovirus(GPV), Duck paramyxovirus(DPMV) and Duck hepatitis virus(DHV). The positive rate of RT-PCR method for detecting viral pathogens in 8 field NDRV isolates and 3 samples of the liver and spleen tissues of ducklings which were artificially infected by the strain of NDRV-NP03 was 100%. The results indicate that this RT-PCR method is sensitive and specific for detecting NDRV, and can be used in NDRV clinical diagnosis and epidemiology investigation.