Establishment and Performance Evaluation of Method for Detecting Serum Soluble fms Like Tyrosine Kinase-1 Based on Biotin-avidin System Time-resolved Fluorescence Immunoassay
10.3969/j.issn.1671-7414.2025.02.035
- VernacularTitle:基于生物素亲和素时间分辨荧光免疫法检测血清可溶性fms样酪氨酸激酶-1的方法建立及性能评价
- Author:
Yuhua TAN
1
;
Ting YU
;
Haijia YU
;
Gaocheng LI
;
Jianming FENG
Author Information
1. 广州市丰华生物股份有限公司体外诊断试剂研发中心,广州 510730
- Publication Type:Journal Article
- Keywords:
biotin;
avidin;
time-resolved fluorescence immunoassay;
soluble fms-like tyrosine kinase-1;
performance evaluation
- From:
Journal of Modern Laboratory Medicine
2025;40(2):186-190,201
- CountryChina
- Language:Chinese
-
Abstract:
Objective To establish a biotin-avidin system time-resolved fluorescence immunoassay(BAS-TRFIA)for detecting serum soluble fms-like tyrosine kinase-1(sFlt-1)and evaluate its performance.Methods A BAS-TRFIA was established to quantitatively determine the concentration of sFlt-1 in the serum of pregnant women,which based on the microplate was coated with streptavidin.The monoclonal antibody to capture sFlt-1 was labeled by biotin,and the detection of sFlt-1 monoclonal antibody was labeled by europium.The performance indicators such as lower limit of detection,biological limit of detection,functional sensitivity,precision,linearity,interference test,cross-reaction test,and high dose hook effect of the method were evaluated.A total of 106 remaining serum samples from pregnant women with no hemolysis,jaundice and lipemia at more than 9 weeks were detected by BAS-TRFIA and electrochemiluminescence for methodological comparison study,and the correlation of the comparison test results was analyzed by linear regression.Results The optimal reaction time of the sample was 90 min.The lower limit of detection was 1.00pg/ml.The biological limit of detection was 10.00pg/ml,and the functional sensitivity was 10.00pg/ml.The intra-assay CV and inter-assay CV were both within 5%,and the linear range was 20.00 to 40 000.00pg/ml.The relative bias of the detection results of the 17 interfering samples with interfering substances added to the low-concentration and high-concentration quality controls and the basic samples was within-4.94%～4.24%.The high dose hook effect was not found in sFlt-1 samples up to 150 000pg/ml.When the concentration of sFlt-1 in the sample was 105.40～40 972.00pg/ml,the linear regression equation of BAS-TRFIA and electrochemiluminescence(ECL)detection results was Y=1.086 7X+17.946(r=0.994 4,t=96.26,P＜0.05).Conclusion The quantitative detection of sFlt-1 by BAS-TRFIA has high sensitivity,good precision,wide linear range,strong anti-interference ability,and good correlation with the detection results of reference methods,which is valuable for clinical application.
