Abstract: An indirect ELISA was developed using the purified recombinant novel duck reovirus(NDRV)σB and σC proteins as coating antigens. The optimized conditions for the methodology were determined to include:concentrations of σB at 12.5 μg·mL^-1 and σC at 6.25 μg·mL^-1,a blocking buffer of 15% FBS, serum samples being diluted 80 times, goat anti-duck HRP-IgG being diluted 400 times,and substrate incubation at 37℃ for 5 min. The assay was found to be specific without any cross reaction with antibodies of MDRV, DHV, MPV, or MD-GPV. The coincidence rate between the indirect ELISA coated with the recombinant σB and σC proteins and that coated with NDRVwas 92.5%. It appeared that the newly developed ELISA exhibited satisfactory sensitivity and specificity on measurements,and could be an alternative means for detecting anti-NDRV antibodies.