[1]谭玉华,于 婷,余海枷,等.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(02):186.[doi:10.3969/j.issn.1671-7414.2025.02.035]
　TAN Yuhua,YU Ting,YU Haijia,et al.Establishment and Performance Evaluation of Method for Detecting Serum Soluble fms Like Tyrosine Kinase-1 Based on Biotin-avidin System Timeresolved Fluorescence Immunoassay[J].Journal of Modern Laboratory Medicine,2025,40(02):186.[doi:10.3969/j.issn.1671-7414.2025.02.035]
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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()

《现代检验医学杂志》[ISSN:/CN:]

卷:
第40卷
期数:
2025年02期
页码:
186
栏目:
研究简报·实验技术
出版日期:
2025-03-15

文章信息/Info

Title:
Establishment and Performance Evaluation of Method for Detecting Serum Soluble fms Like Tyrosine Kinase-1 Based on Biotin-avidin System Timeresolved Fluorescence Immunoassay
文章编号:
1671-7414（2025）02-186-06
作者:
谭玉华1，于 婷2，余海枷1，李高成1，冯健明1
（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）
Author(s):
TAN Yuhua1，YU Ting2，YU Haijia1，LI Gaocheng1，FENG Jianming1
（1. R & D Center of IVD Reagents， Guangzhou Fenghua Biotech Co.Ltd，Guangzhou 510730，China；2. Institute for in Vitro Diagnostic Control， National Institutes for Food and Drug Control，Beijing 100050，China）
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
分类号:
R446
DOI:
10.3969/j.issn.1671-7414.2025.02.035
文献标志码:
A
摘要:
目的 建立检测血清可溶性fms 样酪氨酸激酶-1（sFlt-1）的生物素- 亲和素时间分辨荧光免疫法（BAS-TRFIA），并对其性能进行评价。方法 利用链霉亲和素包被微孔板、生物素标记捕获sFlt-1 单克隆抗体和铕标记检测sFlt-1 单克隆抗体，建立一种BAS-TRFIA 定量测定孕妇血清中的sFlt-1 浓度。对该方法的检测低限、生物检测限、功能灵敏度、精密度、线性、干扰试验和高剂量钩状效应等性能指标进行评价。选择106 份孕周大于9 周的无溶血、黄疸和脂血的孕妇血清剩余样本，用于BAS-TRFIA 和电化学发光法的比对研究，比对试验结果的相关性采用线性回归分析。结果 该方法的样本最适反应时间为90min，检测低限为1.00pg/ml，生物检测限为10.00pg/ml，功能灵敏度为10.00pg/ml，批内和批间变异系数均在5.00% 以内，线性范围为20.00 ～ 40 000.00pg/ml，分别在低浓度和高浓度质控品中添加有干扰物的17 种干扰样本与基础样本检测结果的相对偏倚在-4.94% ～ 4.24% 内；检测样品sFlt-1 浓度高达150 000pg/ml 时未出现高剂量钩状效应；样本sFlt-1 浓度在105.40 ～ 40 972.00pg/ml 间，与电化学发光法（ECL）检测结果的线性回归方程为Y=1.086 7X+17.946（r =0.994 4，t=96.26，P ＜ 0.05）。结论 BAS-TRFIA 具有灵敏度高、精密度好、线性范围较宽和抗干扰能力强，与参比方法检测结果的相关性良好等优点，可以满足临床需要。
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.

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备注/Memo

备注/Memo:
作者简介： 谭玉华（1980-），男，医学硕士，医疗器械高级工程师，临床医学检验技师，二级企业培训师，三级健康管理师，研究方向：医疗器械（含体外诊断试剂）的研发、注册与应用，临床医学检验和标准化研究，E-mail: tanywhy@aliyun.com。
于婷（1978-），女，博士，研究员，研究方向：体外诊断试剂质量控制与评价、标准化研究以及标准物质研制，E-mail：yuting@nifdc.org.cn。并列第一作者。
更新日期/Last Update: 2025-03-15