[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;spec...[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 10μg/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∶12000 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 10μg/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∶12000 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.
