[1]王星皓,胡高峰,许成山,等.临床实验室血浆抗凝蛋白S活性检测日间精密度与凝血因子Ⅷ，Ⅸ定量限的性能验证方法学评估及改善方案研究[J].现代检验医学杂志,2025,40(02):195-201.[doi:10.3969/j.issn.1671-7414.2025.02.037]
　WANG Xinghao,HU Gaofeng,XU Chengshan,et al.Study on the Methodological Evaluation and Improvement for the Performance Verification of Inter-day Precision in Plasma Anticoagulant Protein S Activity Assay and Limits of Quantitative of Coagulation Factors Ⅷ and Ⅸ[J].Journal of Modern Laboratory Medicine,2025,40(02):195-201.[doi:10.3969/j.issn.1671-7414.2025.02.037]
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临床实验室血浆抗凝蛋白S活性检测日间精密度与凝血因子Ⅷ，Ⅸ定量限的性能验证方法学评估及改善方案研究()
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《现代检验医学杂志》[ISSN:/CN:]

卷:
第40卷
期数:
2025年02期
页码:
195-201
栏目:
质量控制·实验室管理
出版日期:
2025-03-15

文章信息/Info

Title:
Study on the Methodological Evaluation and Improvement for the Performance Verification of Inter-day Precision in Plasma Anticoagulant Protein S Activity Assay and Limits of Quantitative of Coagulation Factors Ⅷ and Ⅸ
文章编号:
1671-7414（2025）02-195-07
作者:
王星皓1; 2; 胡高峰1; 许成山1; 彭明婷1; 2
（1. 北京医院/ 国家老年医学中心，国家卫生健康委临床检验中心，中国医学科学院老年医学研究院，北京 100730；2. 北京协和医学院/ 中国医学科学院，北京 100730）
Author(s):
WANG Xinghao1; 2; HU Gaofeng1; XU Chengshan1; PENG Mingting1; 2
(1. Beijing Hospital / National Center of Gerontology，National Center for Clinical Laboratories，Institute of Geriatric Medicine，Chinese Academy of Medical Sciences，Beijing 100730，China；2. Chinese Academy of Medical Sciences & Peking Union Medical College， Beijing 100730，China)
关键词:
抗凝蛋白S; 凝血因子; 日间精密度; 定量限; 性能验证
分类号:
R446
DOI:
10.3969/j.issn.1671-7414.2025.02.037
文献标志码:
A
摘要:
目的 针对抗凝蛋白S（PS）、凝血因子Ⅷ活性（F Ⅷ :C）和凝血因子Ⅸ活性（F Ⅸ :C）检测性能验证中存在的问题，探讨PS 活性检测日间精密度验证的改进方案，以及F Ⅷ :C 和F Ⅸ :C 检测定量限（LoQ）的验证方案。方法使用Sysmex CN-6000 及配套试剂，使用低值和高值质控品作为研究样品。参考美国临床和实验室标准协会（CLSI）EP15-A3 文件，使用三种试剂准备方式（说明书方式、仪器操作手册方式和改进方式）进行PS 活性检测日间精密度验证研究。说明书要求方式即完全参照产品说明书开展；仪器操作手册方式在其基础上将所需试剂于仪器中静置30min后用于检测；改进方式在产品说明书基础上对所需试剂进行混合分装。研究同一批号试剂检测同一样品的瓶间差异，以国家卫生健康委临床检验中心（NCCL）室间质评（EQA）可接受范围作为评价标准。参考CLSI EP25 文件，验证产品说明书方式与改进方式复溶试剂的机载稳定性。参考CLSI EP17-A2 文件、WS/T 514-2017《临床检验方法检出能力的确立和验证》和国际血液学标准化委员会（ICSH）指南，进行F Ⅷ :C 和F Ⅸ :C 检测LoQ 验证。研究结果均以满足产品说明书要求为验证通过。结果 参照产品说明书与仪器操作手册进行PS 活性检测精密度验证，日间精密度（CV日间：12.9% ～ 21.6%）均超出产品说明书要求（高值水平的CV批内，CV日间均＜ 10%；低值水平的CV批内，CV日间均＜ 20%）。应用改进方式的日间精密度验证结果（CV日间：2.9% 和4.5%）符合产品说明书要求。试剂瓶间差异超出EQA 可接受范围。改进方式纠正了试剂机载稳定性验证结果（相对偏差：-4.24% ～ 9.97%）符合产品说明书（高值水平＜ 10%，低值水平＜ 20%）。F Ⅷ :C 和F Ⅸ :C 检测的LoQ 验证通过范围均符合厂商声明（F Ⅷ :C：0.75% ～ 1.46%，0.74% ～ 1.40%；F Ⅸ :C：0.71% ～ 1.27%，0.70% ～ 1.32%）。结论 提供了改善PS 活性检测日间精密度的改进方式，并提供了F Ⅷ :C和F Ⅸ :C 检测的LoQ 验证方案，以供临床实验室参考。
Abstract:
Objective To discuss the improvement for the verification of inter-day precision of PS activity assay and the verification protocol for limits of quantitative of F Ⅷ :C and F Ⅸ :C assay, aiming at the problems in the performance verification of anticoagulant protein S (PS), coagulation factor Ⅷ activity (F Ⅷ :C) and coagulation factor Ⅸ activity (F Ⅸ :C) assay. Methods SYSMEX CN-6000 and supporting reagents were used, and low- and high-value quality control (QCs) were used as the study samples. Following the American Clinical and Laboratory Standards Institute (CLSI) document EP15-A3, an inter-day precision verification study of the PS activity assay was performed with three reagent preparation methods designed (specification-required method, instrument’s manual method and improved method). The specification-required method was carried out completely according to the specification, and the instrument manual method involved allowing the required reagents to stand in the device for 30 minutes based on the specification-required method, and the improved method mixed and dispensed the reagents needed to be based on the specification-required method. To study the bottle variation of the same batch of reagents for the same sample, the acceptable range of external quality assessment (EQA) of the National Center for Clinical Laboratories (NCCL) was used as the evaluation standard. Following the CLSI EP25 document, the PS activity assay reagent onboard stability verification was performed with the specification-required method and the improved method. Following the CLSI EP17-A2 document, WS/T 514-2017 “Establishment and Verification of Detection Capability for Clinical Laboratory Measurement Procedures” and the International Committee for Standardization in Hematology (ICSH) guidelines, the LoQ for FVIII: C and FIX: C assays was verified. The verification results were passed if the requirements of the specification were met. Results The verification result of inter-day precision (CVWL：12.9% ～ 21.6%) of PS activity assay according to the specification method and the instrumental manual method exceeded the requirements of the specification (<10% CVWL at high levels, <20% CVWL at low levels). The results of inter-day precision verification with the improved method (CVWL：2.9% and 4.5%) were consistent with the requirements of the specification. The bottle variation exceeded the acceptable range of EQA. The improved method corrected the reagent on-board stability verification results of reagents (relative deviations of -4.24% ～ 9.97%) by the specifications (high levels <10%, low levels <20%). The LoQ verification results for F Ⅷ :C and F Ⅸ :C assay were by the product specifications (F Ⅷ :C：0.75% ～ 1.46% and 0.74% ～ 1.40%；F Ⅸ :C：0.71% ～ 1.27% and 0.70% ～ 1.32%). Conclusion An improved method to improve the inter-day precision of PS activity detection is provided, and LoQ verification protocol for F Ⅷ :C and F Ⅸ :C assay is provided for clinical laboratory reference.

参考文献/References:

[1] GARDINER C, COLEMAN R, DE MAAT M P M, et al. International Council for Standardization in Haematology (ICSH) laboratory guidance for the evaluation of haemostasis analyser-reagent test systems. Part 1: Instrument-specific issues and commonly used coagulation screening tests [J]. International Journal of Laboratory Hematology, 2021, 43(2):169-183.
[2] GARDINER C, COLEMAN R, DE MAAT M P M, et al. International Council for Standardization in Haematology (ICSH) laboratory guidance for the verification of haemostasis analyser-reagent test systems. Part 2: Specialist tests and calibrated assays [J].International Journal of Laboratory Hematology, 2021, 43(5): 907-916.
[3] 罗国菊, 寿炜龄, 陈倩, 等.CN-3000 全自动血凝仪检测凝血七项的性能验证[J].标记免疫分析与临床,2022,29(7):1204-1208. LUO Guoju, SHOU Weiling, CHEN Qian, et al. A performance verification of CN-3000 automatic blood coagulationanalyzer for seven routine blood coagulation tests [J]. Labeled Immunoassays and Clinical Medicine, 2022, 29(7): 1204-1208.
[4] 侯军林, 赵旭宏, 张曼. 两种血凝分析仪检测系统性能验证及结果比对[J]. 中华医学杂志,2011,91(16): 1139-1142. HOU Junlin, ZHAO Xuhong, ZHANG Man. Comparison and evaluation of testing results for two different coagulation analyzers[J] . National Medical Journal of China, 2011, 91(16): 1139-1142.
[5] 宋文良, 许巍. 儿童易栓症的诊断思路[J]. 中国小儿急救医学,2023,30(10):736-740. SONG Wenliang, XU Wei. Diagnostic thought of pediatric thrombophilia [J] .Chinese Pediatric Emergency Medicine, 2023,30(10):736-740.
[6] 中华医学会血液学分会血栓与止血学组. 易栓症诊断与防治中国指南(2021 年版)[J]. 中华血液学杂志,2021,42(11):881-888. Thrombosis and Hemostasis Group,Chinese Society of Hematology,Chinese Medical Association. Chinese guidelines for diagnosis,prevention and treatment of thrombophilia(2021) [J].Chinese Journal of Hematology, 2021,42(11):881-888.
[7] MIDDELDORP S, NIEUWLAAT R, BAUMANN KREUZIGER L,et al. American society of hematology 2023 guidelines for management of venous thromboembolism: thrombophilia testing [J]. Blood Advances, 2023, 7(22): 7101-7138.
[8] MARLAR R A, GAUSMAN JN, TSUDA H, et al. Recommendations for clinical laboratory testing for protein S deficiency: communication from the SSC committee plasma coagulation inhibitors of the ISTH [J]. Journal of Thrombosis and Haemostasis, 2021, 19(1): 68-74.
[9] 曹修娥, 苏艳, 贾传华, 等.HemoCELL 全自动血凝流水线检测抗Xa?蛋白C 和蛋白S 活性的性能评价[J]. 中国医学装备,2022,19(1):53-56. CAO Xiu’e, SU Yan, JIA Chuanhua, et al. Performance evaluation of HemoCELL automation coagulation assembly line in detecting the activities of anti-Xa, P-C and P-S[J].China Medical Equipment, 2022, 19(1): 53-56.
[10] 刘秀丽, 周文宾, 李臣宾, 等. 抗凝蛋白检测现状全国性调查与分析[J].中华医学杂志,2017,97(22):1699-1704. LIU Xiuli, ZHOU Wenbin, LI Chenbin, et al. Study and analysis of National current status and problems of anticoagulant proteins assay[J]. National Medical Journal of China, 2017, 97(22): 1699-1704.
[11] 虞红燕, 刘禹, 郁大伟, 等.2015 ~ 2017 年上海地区人群血浆PC 和PS 活性检测情况的调查分析[J].现代检验医学杂志,2019,34(4):154-157. YU Hongyan, LIU Yu, YU Dawei, et al. Investigation and analysis of plasma PC and PS activities in Shanghai population from 2015 to 2017[J]. Journal of Modern Laboratory Medicine, 2019, 34(4): 154-157.
[12] VAN MOORT I, MEIJER P, PRIEM-VISSER D, et al. Analytical variation in factor VIII one-stage and chromogenic assays: Experiences from the ECAT external quality assessment programme[J].Haemophilia, 2019, 25(1): 162-169.
[13] 刘禹, 许冠群, 戴菁, 等. 凝血因子Ⅷ活性检测现状及发展趋势[J]. 中华检验医学杂志,2022,45(10):1010-1016. LIU Yu, XU Guanqun, DAI Jing, et al. Current status and development trend of coagulation factor Ⅷ activity[J]. Chinese Journal of Laboratory Medicine, 2022, 45(10): 1010-1016.
[14] 中华医学会血液学分会血栓与止血学组, 中国血友病协作组. 血友病诊断与治疗中国专家共识(2017年版)[J]. 中华血液学杂志,2017,38(5):364-370. Thrombosis and Hemostasis Group, Hematology Society of Chinese Medical Association, Hemophilia Treatment Center Collaborative Network of China. Consensus of Chinese experts on the diagnosis and treatment of hemophilia (version 2017) [J]. Chinese Journal of Hematology, 2017, 38(5): 364-370.
[15] 李敏, 寿玮龄, 范连凯, 等. 一期凝固法凝血因子Ⅷ活性检测医学决定水平的性能验证[J]. 标记免疫分析与临床,2022,29(6):1015-1020. LI Min, SHOU Weiling, FAN Liankai, et al. The validation of medical determination level of coagulation factor Ⅷ activity by one-stage coagulation method[J].Labeled Immunoassays and Clinical Medicine, 2022, 29(6): 1015-1020.
[16] Clinical and Laboratory Standards Institute. Evaluation of detection capabilityfor clinical laboratory measurement procedures; approved guideline-Second edition [S].Wayne:PA,CLSI EP17-A2, 2012.
[17] Clinical and Laboratory Standards Institute. User verification of precision and estimation of bias; approved guideline-Third edition [S]. Wayne:PA,CLSI EP15-A3, 2014.
[18] GOMEZ-RIOJA R, VON MEYER A, CORNES M, et al. Recommendation for the design of stability studies on clinical specimens[J]. Clinical Chemistry and Laboratory Medicine, 2023, 61(10): 1708-1718.
[19] Clinical and Laboratory Standards Institute. Evaluation of stability of in vitro medical laboratory test reagents-Second edition [S]. Wayne:PA,CLSI EP25, 2023.
[20] 中华人民共和国国家卫生和计划生育委员会.WS/T514-2017: 临床检验方法检出能力的确立和验证 [S].北京: 中国标准出版社,2017. National Health and Family Planning Commission of the People’s Republic of China. WS/T 514-2017: Establishment and verification of detection capability for clinical laboratory measurement procedures [S]. Beijing: China Standard Publishing House, 2017.
[21] Sysmex CN-3000 and CN-6000 application guide. Fully Automated Coagulation Analysers CN-3000/CN-6000 [Z].( 2019-02) http:// www.sysmex-ap.com 2019:2

备注/Memo

备注/Memo:
基金项目：国家自然科学基金（81772254）。
作者简介：王星皓（2000-）, 男, 硕士在读, 研究方向：临床血液学检验标准化,E-mail:wxh1050@foxmail.com。
通讯作者：彭明婷, 研究员, 主要从事临床血液、体液学检验质量控制和标准化研究,E-mail:mtpeng@nccl.org.cn。
更新日期/Last Update: 2025-03-15