The development and application of a blocking ELISA kit for the diagnosis of infectious coryza

Abstract

A blocking enzyme-linked immunosorbent assay (B-ELISA) kit for the diagnosis of infectious coryza was developed in this study. The kit was based on a recently described blocking ELISA that uses monoclonal antibodies to achieve specificity for antibodies to either Haemophilus paragallinarum serovar A or serovar C. The results showed that the B-ELISA kit detected 96 and 90%, respectively, of chickens vaccinated or challenged with H. paragallinarum serovar A. When used on chickens vaccinated or challenged with H. paragallinarum serovar C, the kit detected 77 and 40%, respectively, as positive. The majority of sera from vaccinated chickens were still positive on the serovars A and C ELISAs 4 months after vaccination. Based on pen trial data, the serovar A B-ELISA kit had a sensitivity of 95% and a specificity of 100%. The serovar C B-ELISA kit had a sensitivity of 73% and a specificity of 100%. A range of field sera was examined with the kit, generating results that correlated with the known vaccination/disease history of the flocks examined. As freeze drying the monoclonal antibodies and the conjugate had some effect on optimal working concentration, the kit used liquid solutions of these two reagents. The kit could be stored for 7 days at 37 deg C, 10 months at 4 deg C and more than 1 year at -20 deg C. Our results suggest that the kit would be a useful aid in the diagnosis of infectious coryza in China and other countries where H. paragallinarum serovars A and C are the predominant or sole serovars.