[Objective]The aim was to establish an indirect ELISA for detection on antibody of Hps. [Method] The optimal conditions of indirect ELISA were selected and determined based on heat-resistant serotype 4 and 5 of Hps; s...[Objective]The aim was to establish an indirect ELISA for detection on antibody of Hps. [Method] The optimal conditions of indirect ELISA were selected and determined based on heat-resistant serotype 4 and 5 of Hps; specific,repeating and sensitive tests were conducted and 200 serums were detected. [Result]The optimal conditions were as follows： coating concentration of antigen at 10 μg /ml,and coating for 2 h at 37 °C; PBST containing 20 g /L of skim milk powder as blocking fluid for 30 min; serum dilution at 1∶ 80; reaction time of antigen for 45 min; dilution of secondary antibody at 1∶ 12 000 and effecting for 30 min; color development reaction for 15 min. [Conclusion] The established indirect ELISA is good in specificity and repetitiveness with higher sensitivity than that of indirect hemagglutination test; the results of clinic samples （ negative /positive serums） were in consistent with those detected with foreign ELISA kits. The established method can be made use of in serum antibody of Hps de- tection and seroepidemiology study.展开更多
基金funded by Scientific Research Foundation of Guangdong Dahuanong Animal Health Products CO. LTD.(033A201055-2)
文摘[Objective]The aim was to establish an indirect ELISA for detection on antibody of Hps. [Method] The optimal conditions of indirect ELISA were selected and determined based on heat-resistant serotype 4 and 5 of Hps; specific,repeating and sensitive tests were conducted and 200 serums were detected. [Result]The optimal conditions were as follows： coating concentration of antigen at 10 μg /ml,and coating for 2 h at 37 °C; PBST containing 20 g /L of skim milk powder as blocking fluid for 30 min; serum dilution at 1∶ 80; reaction time of antigen for 45 min; dilution of secondary antibody at 1∶ 12 000 and effecting for 30 min; color development reaction for 15 min. [Conclusion] The established indirect ELISA is good in specificity and repetitiveness with higher sensitivity than that of indirect hemagglutination test; the results of clinic samples （ negative /positive serums） were in consistent with those detected with foreign ELISA kits. The established method can be made use of in serum antibody of Hps de- tection and seroepidemiology study.
