A method of human serum folic acid dectetion by non-equilibrium competitive immunoassay using FITC detecting system / 重庆医学

ABSTRACT

Objective To prepare anti-folic acid (FA) polyclonal antibody and develop a new non-balanced competing chemiluminescence analysis for clinical detection of FA.Methods Established the detection method by added FITC-FA-analogs and FAHRP-antibody in the light emitting plate,which coated with anti-FITC antibody,to form the immune response complex of FITC/antibody-FITC-FA-analogs/FA-antibody-HRP.Then methodology evaluation was performed to evaluate the method performance;and further compared the detecting results with non-FITC system detection system and Electrochemiluminescence system (Roche Elecsys 2010).Results The ELISA results showed that the prepared anti-FA antibodies can recognize serum FA specificly.The methodology evaluation indicated that the linear correlation coefficient of the standard curve was 0.990 0;the analytical sensitivity was 1.21 ng/mL;the range of linear detection was 1.21～ 38.80 ng/mL;The coefficient variability of intra-assay was ＜5 ％,which was better than the results of non-FITC detection system;and the correlation coefficient was 0.908 1 compared with the Elecsys-2010 detection system.Conclusion The established chemiluminescence immunoassay for human serum FA has a good sensitivity and specificity,and suitable for clinical serum FA quantitativedetecting.