Development of Indirect ELISA for Detecting Novel Duck Reovirus Using σB and σC Proteins as Coating Antigens

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.