[1]谭玉华,冯健明,于 婷,等.全自动时间分辨荧光免疫分析技术检测血清抑制素A的方法建立及性能评价[J].现代检验医学杂志,2026,41(02):23-28+45.[doi:10.3969/j.issn.1671-7414.2026.02.005]
　TAN Yuhua,FENG Jianming,YU Ting,et al.Establishment and Performance Evaluation of Automatic Time Resolved Fluorescence Immunoassay for the Detection of Serum Inhibin A[J].Journal of Modern Laboratory Medicine,2026,41(02):23-28+45.[doi:10.3969/j.issn.1671-7414.2026.02.005]
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全自动时间分辨荧光免疫分析技术检测血清抑制素A的方法建立及性能评价()

《现代检验医学杂志》[ISSN:/CN:]

卷:
第41卷
期数:
2026年02期
页码:
23-28+45
栏目:
出版日期:
2026-03-15

文章信息/Info

Title:
Establishment and Performance Evaluation of Automatic Time Resolved Fluorescence Immunoassay for the Detection of Serum Inhibin A
文章编号:
1671-7414（2026）02-023-07
作者:
谭玉华1，冯健明1，于 婷2，余海枷1，陈 龙1，戴林彬1
1.广州市丰华生物股份有限公司，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，FENG Jianming1，YU Ting2，YU Haijia1，CHEN Long1，DAI Linbin1
1.Guangzhou Fenghua Bioengineering Co.Ltd., Guangzhou 510730, China;2.Institute for in Vitro Diagnostic Control, National Institutes for Food and Drug Reagents Testing, Beijing 100050, China
关键词:
时间分辨荧光免疫分析; 抑制素A; 性能评价
分类号:
R446.11；Q503
DOI:
10.3969/j.issn.1671-7414.2026.02.005
文献标志码:
A
摘要:
目的 建立一种全自动时间分辨荧光免疫分析(TRFIA)技术检测血清抑制素A(INHA)的方法,并对其性能进行评价。方法抑制素βA单克隆抗体包被微孔板作为微孔反应板,铕标记抑制素α单克隆抗体作为铕标记物,采用全自动TRFIA仪定量检测孕妇血清中INHA浓度,对其分析灵敏度、准确度、回收率、线性、精密度、校准品均匀性和特异度等性能指标进行评价。选择69例孕周在14~21周的无溶血、黄疸和脂血的孕妇血清样本同时进行化学发光法及TRFIA法检测进行方法学比对试验,一致性采用t检验、线性相关分析、回归分析和Bland-Altman法分析。结果抑制素βA单克隆抗体最适包被浓度为4μg/ml,铕标记抑制素α单克隆抗体母液最适稀释比为1:1000,检测低限、生物检测限和功能灵敏度分别为0.75、10.00和25.00pg/ml,检测国际标准品的相对偏差在-9.28%~-2.17%,回收试验的平均回收率在92.71%~100.79%。在20.00~2000.00pg/ml范围内,线性相关系数可达0.9995,批内和批间变异系数(CV)分别在1.56%~1.71%和1.61%~2.22%,校准品B~F的瓶间CV在2.23%~3.47%。交叉反应中,检测高浓度的人促甲状腺激素(hTSH)、人促黄体激素(hLH)、泌乳素(PRL)、人促卵泡激素(hFSH)、人绒毛膜促性腺激素(hCG)、人甲胎蛋白(hAFP)、α2-巨球蛋白(AMG)、激活素A(ACTA)、激活素AB(ACTAB)、激活素B(ACTB)、抑制素B(INHB)和卵泡抑素(FST),结果均不高于10pg/ml。自建方法与对比方法定量测定结果高度相关(r=0.9968,tr=101.99,P<0.05),95.65%的相对偏差数据点在95%一致性限(-7.48%,7.63%)内,且95%一致性限的置信区间(-9.07%,9.24%)在±10%内。结论自建方法具有灵敏度高、准确度好、线性范围宽、精密度好和特异度高等优点,与对比方法检测结果具有高度的相关性和一致性,满足临床检测需求。
Abstract:
Objective To establish an automatic time-resolved fluorescence immunoassay (TRFIA) for the detection of serum inhibin A (INH A) and evaluate its performance. Methods The microwell plate was coated with anti-inhibin βA monoclonal antibody served as microwell reaction plate. Europium-labeled anti-inhibin α monoclonal antibody was used as the europium marker. The INH A concentrations in maternal serum were quantitatively detected by automatic TRFIA instrument. Perfor-mance indicators including analytic sensitivity, accuracy, recovery rate, linearity, precision, calibration uniformity, and specific-ity were evaluated. A total of 69 serum samples from pregnant women without hemolysis, jaundice and lipemia were selected from 14 to 21 weeks of gestation and tested concurrently using chemiluminescence and TRFIA methods for methodological comparison. The consistency was analyzed by t test, linear correlation analysis, regression analysis and the Bland-Altman method. Results The optimal coating concentration for anti-inhibin βA monoclonal antibody was 4 μg/ml. The optimal di-lution ratio for the parent solution of europium-labeled anti-inhibin α monoclonal antibody was 1:1 000. The detection lim-it, biological detection limit and functional sensitivity were 0.75, 10.00 and 25.00 pg/ml, respectively. The relative deviation for international standards ranged from -9.28% to -2.17%, and the average recovery rate was from 92.71% to 100.79%.The linear correlation coefficient reached 0.999 5 within the range of 20.00 to 2000.00 pg/ml. The intra-assay and inter-assay co-efficients of variation ranged from 1.56% to 1.71% and 1.61% to 2.22%, respectively. The bottle-to-bottle coefficient of variation for calibrators B ~ F ranged from 2.23% to 3.47%. In the cross-reactivity testing, high concentrations of human thyrotropin (hTSH), human luteinizing hormone (hLH), prolactin (PRL), human follicle-stimulating hormone (hFSH), human chorionic go-nadotropin (hCG), human alpha-fetoprotein (hAFP), α2-macroglobulin (AMG), activin A (ACTA), activin AB (ACTAB), activin B (ACTB), inhibin B (INH B) and folliclestatin (FST) were detected, and the results were not higher than 10 pg/ml. The quantita-tive results of the self-developed method were highly correlated with those of the reference method (r= 0.996 8, tr=101.99, P <0.05). 95.65% of the relative deviation data points fell within the 95% consistency limits (-7.48% to 7.63%), and the 95% confi-dence interval of the consistency limits (-9.07% to 9.24%) fell within ±10%. Conclusions The self-developed method has the advantages of high sensitivity, good accuracy, a wide linear range, good precision, and high specificity, which has high correlation and consistency with the reference method, meeting clinical testing requirement.

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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。
冯健明（1978-），男，本科，主管检验技师，研究方向：医疗器械（含体外诊断试剂）的研发、注册与应用，E-mail:845776124@qq.com，并列第一作者。
通讯作者： 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。
更新日期/Last Update: 2026-03-15