[1]谭玉华,于 婷,余海枷,等.Auto TRFIA-4型全自动荧光免疫分析仪血片支架固定滤纸血斑检测17α-羟孕酮的试剂研制及性能评价[J].现代检验医学杂志,2025,40(05):182-188.[doi:10.3969/j.issn.1671-7414.2025.05.035]
　TAN Yuhua,YU Ting,YU Haijia,et al.Development and Performance Evaluation of A Reagent for Detecting of 17α-hydroxyprogesterone in Dried Blood Spots on Filter Paper Fixed on Blood Spots Stent by Auto TRFIA-4 Automatic Fluorescence Immunoanalyzer[J].Journal of Modern Laboratory Medicine,2025,40(05):182-188.[doi:10.3969/j.issn.1671-7414.2025.05.035]
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Auto TRFIA-4型全自动荧光免疫分析仪血片支架固定滤纸血斑检测17α-羟孕酮的试剂研制及性能评价()

《现代检验医学杂志》[ISSN:/CN:]

卷:
第40卷
期数:
2025年05期
页码:
182-188
栏目:
研究简报·实验技术
出版日期:
2025-09-15

文章信息/Info

Title:
Development and Performance Evaluation of A Reagent for Detecting of 17α-hydroxyprogesterone in Dried Blood Spots on Filter Paper Fixed on Blood Spots Stent by Auto TRFIA-4 Automatic Fluorescence Immunoanalyzer
文章编号:
1671-7414（2025）05-182-07
作者:
谭玉华1，于 婷2，余海枷1，陈梅欣1，谢敬玲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，CHEN Meixin1，XIE Jingling1，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:
时间分辨荧光免疫分析法; 血片支架; 滤纸血斑; 17α-羟孕酮
分类号:
R446
DOI:
10.3969/j.issn.1671-7414.2025.05.035
文献标志码:
A
摘要:
目的研制一种AutoTRFIA-4型全自动荧光免疫分析仪血片支架固定滤纸血斑检测17α-羟孕酮（17α-OHP）的试剂，并对其性能进行评价。方法羊抗兔IgG抗体包被微孔板作为微孔反应板，稀释后的兔抗人17α-OHP抗体作为中间抗体，铕标记17α-OHP-牛血清白蛋白偶联物作为铕标记物，采用AutoTRFIA-4型全自动荧光免疫分析仪血片支架固定滤纸血斑定量检测17α-OHP浓度，对其分析灵敏度、准确度、线性、精密度、特异度和稳定性等性能指标进行评价，并评估是否满足拟订的行业标准的要求。选择227份出生72h后、7天内、并充分哺乳的新生儿足跟血滤纸干血片样本用于试剂比对试验，一致性采用χ2检验、Kappa检验、t检验、线性相关分析、回归分析、Bland-Altman法分析和医学决定水平预期偏倚分析。结果羊抗兔IgG抗体最适包被浓度为3μg/ml，兔抗人17α-OHP抗体最适稀释比为1∶1500，17α-OHP铕标记物母液最适稀释比为1∶2500；空白限、检出限和定量限分别为0.75，1.08和1.99nmol/L；标准品核对试验的相对偏差在±15.00%范围内，回收试验的平均回收率为92.36%；在2.00～300.00nmo/L范围内，线性相关系数为0.9971；批内和批间变异系数均在10.00%以内；检测100.00ng/ml的孕酮、17α-羟孕烯醇酮、11-脱氧皮质醇的样品，交叉反应率在0.089%～0.64%范围内。稳定性试验的性能符合要求。与对比试剂检测结果的总符合率为100%，与对比试剂定量测定结果高度相关（r=0.9994，tr=452.02，P＜0.05）。结论自研试剂具有灵敏度高、准确度好、线性范围宽、精密度好、特异度高和稳定性好等优点，符合拟订的行业标准要求，与对比试剂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 develop a reagent for detecting of 17α-hydroxyprogesterone （17α-OHP）in dried blood spots on filter paper fixed on vascular stent by Auto TRFIA-4 automatic fluorescence immunoanalyzer and evaluate its performance. Methods The microwell plate was coated with the sheep anti-rabbit IgG antibody as microwell reaction plate,the rabbit anti-human 17α-OHP antibody was diluted as intermediate antibody,and the 17α-OHP- bovine serum albumin conjugate was labeled by europium as europium marker. The concentration of 17α-OHP in dried blood spots on filter paper fixed on blood spots stent was quantitatively detected by Auto TRFIA-4 automatic fluorescence immunoanalyzer.The analysis sensitivity, accuracy, linearity, precision, specificity and stability were evaluated,and whether they met the requirements of the formulated industry standards were evaluated. A total of 227 neonatal heel blood filter paper samples from newborns who were born 72 hours after birth and within 7 days and fully breastfeeding were selected for reagent comparison test. The consistency was analyzed by χ2 test, Kappa test, t test, linear correlation analysis,regression analysis,Bland-Altman method analysis and predictive bias analysis of medical decision level, P<0.05 indicated statistically significant difference. Results The optimal coating concentration of sheep anti-rabbit IgG antibody was 3 μg/ml.The optimal dilution ratio of rabbit anti-human 17α-OHP antibody was 1∶1 500.The optimal dilution ratio of 17α-OHP europium marker mother liquor was 1∶2 500.The limit of blank, limit of detection, limit of quantification was 0.75, 1.08 and 1.99 nmol/L, respectively.The relative deviations of the standard check test were within ±15.00%, and the average recovery rate was 92.36%.The linear correlation coefficient was 0.997 1 in the range of 2.00 to 300.00nmo/L.The intra-assay and inter-assay coefficients of variation were all within 10.00%.The cross-reactivity rates of 100.00 ng/ml progesterone, 17α-hydroxypreg nenolone and 11-deoxycortisol were within 0.089% to 0.64%. The performance of stability test met the requirements.The total coincidence rate was 100% compared with the results of contrast reagent. The quantitative results were highly correlated with the contrast reagent (r=0.999 4, tr=452.02, P< 0.05). Conclusion The self-developed reagent has the advantages of high sensitivity, good accuracy, wide linear range, good precision, high specificity and good stability, which meets the requirements of the formulated industry standards ,and has high correlation and consistency with the result of contrast reagent, which meets the needs of clinical detection.

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相似文献/References:

[1]谭玉华,曹春玲,邢晓敏,等.国产Auto TRFIA-4 型自动荧光免疫分析仪与试剂盒检测妊娠相关血浆蛋白A 的性能评价与参考区间的建立[J].现代检验医学杂志,2020,35(04):75.[doi:10.3969/j.issn.1671-7414.2020.04.018]
　TAN Yu-hua,CAO Chun-ling,XING Xiao-min,et al.Performance Evaluation of Domestic Auto TRFIA-4 Model Automatic Fluorescence Immunoassay and Kit for Detection of Pregnancy Associatied Plasma Protein A and Establishment of Reference Interval[J].Journal of Modern Laboratory Medicine,2020,35(05):75.[doi:10.3969/j.issn.1671-7414.2020.04.018]

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: 2025-09-15