Preparation of the Time-resolved Fluoroimmunoassay Kit for Detecting CA125

Objective To prepare a kit for the detection of CA125 in serum by time-resolved fluoroimmunoassay (TRFIA). Method Two anti-CA125 monoclonal antibodies were used to develop the sandwich TRFIA kit for detection of CA125 in serum. Results The working range of kit was 10～600U/ml. The analytical sensitivity was 1.07U/ml. There was no cross-reactivity with CEA,AFP, CA50, CA15-3 and CA19-9. The intra-assay and inter-assay coefficients of variation were 4.5%～8.1% and 6.9%～11.5%, respectively. The TRFIA CA125 reagents can be stored at 4℃ for 1 year and at 37℃ for 7 days. The cutoff value for TRFIA CA125 kit in healthy subjects (n=425) was 37U/ml. The correlation coefficient of blood samples detection results (n=123) between home-made and commercially available CLIA CA125 kit(Roche) was 0.939. Conclusion CA125 TRFIA kit was a valuable diagnostic kit for clinical application with better sensitivity, specificity, and accuracy.