现代检验医学杂志2025，Vol.40Issue(2)：186-190,201,6.DOI:10.3969/j.issn.1671-7414.2025.02.035
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
Establishment and Performance Evaluation of Method for Detecting Serum Soluble fms Like Tyrosine Kinase-1 Based on Biotin-avidin System Time-resolved Fluorescence Immunoassay
谭玉华 1于婷 2余海枷 1李高成 1冯健明1
作者信息
- 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
- 2. 中国食品药品检定研究院体外诊断试剂检定所,北京 100050
- 折叠
摘要
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.关键词
生物素/亲和素/时间分辨荧光免疫法/可溶性fms样酪氨酸激酶-1/性能评价Key words
biotin/avidin/time-resolved fluorescence immunoassay/soluble fms-like tyrosine kinase-1/performance evaluation分类
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谭玉华,于婷,余海枷,李高成,冯健明..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[J].现代检验医学杂志,2025,40(2):186-190,201,6.
