发色底物法检测凝血因子Ⅷ活性的临床应用评价  

Clinical application evaluation of chromogenic substrate assay in detecting coagulation factor Ⅷ activity

作　　者：刘禹 许冠群[1] 王学锋[1] 戴菁[1] Liu Yu;Xu Guanqun;Wang Xuefeng;Dai Jing(Department of Laboratory Medicine,Ruijin Hospital,Shanghai Jiaotong University School of Medicine,Shanghai 200025,China)

机构地区：[1]上海交通大学医学院附属瑞金医院检验科,上海200025

出　　处：《中华检验医学杂志》2020年第8期816-822,共7页Chinese Journal of Laboratory Medicine

基　　金：国家自然科学基金(81770131,81970127)。

摘　　要：目的对发色底物法测定凝血因子Ⅷ(FⅧ)活性进行方法学建立以及临床应用评价。方法选取2018年1月至2019年5月于上海交通大学医学院附属瑞金医院确诊的血友病A患者40例,获得性血友病A患者20例,狼疮抗凝物质阳性患者26例和表观健康者60名,根据美国临床和实验室标准协会(CLSI)相关文件要求,分别对该检测方法的准确度、不精密度、检测下限、线性范围、携带污染率、参考范围、可报告范围等指标进行验证,并与FⅧ活性检测(一期凝固法)进行方法学比较。同时对发色底物法在获得性血友病A和狼疮抗凝物质阳性患者标本的临床应用进行评估。结果两个水平的定值质控品的检测结果均在厂商提供的定值范围内(偏倚为3.93%~6.79%)。批内不精密度变异系数(CV)为1.86%~2.06%(均≤5%),日间不精密度CV为4.83%~6.90%(均≤15%)。灵敏度CV为11.23%(<20%)。试剂盒提供的参考范围(60.00%~168.00%)适用于本实验室。最大稀释度为1∶16。线性范围为5.00%~193.50%(a=1.0243,R^2=1.000)。临床可报告范围为0.50%~387.00%。携带污染率为0.04%。与FⅧ活性检测(一期凝固法)结果一致性高(R^2=0.961)。FⅧ抑制物滴度检测结果与一期凝固法结果一致性高(R^2=0.973)。狼疮抗凝物质阳性伴FⅧ活性(一期凝固法)降低患者的FⅧ∶C结果与一期凝固法结果间的差异有统计学意义(t=9.232,P<0.05)。结论FⅧ活性检测(发色底物法)试剂盒具有优越的检测性能,各方面均呈现出良好的结果,可用于FⅧ活性水平测定,且临床应用范围更广,干扰因素更少。Objective To establish clinical detection of coagulation factorⅧactivity by chromogenic substrate assay and evaluate its clinical application.Methods A total of 40 hemophilia Apatients,20 acquired hemophilia A patients,26 patients with positive lupus anticoagulant and 60 apparently healthy people were enrolled from January 2018 to May 2019 in Ruijin Hospital of Medical College,Shanghai Jiaotong University.According to Clinical and Laboratory Standards Institute(CLSI),the accuracy,within-run and between-run imprecision,lower detection limit,linear range,carryover rate,reference range,and reportable range of chromogenic substrate assay for detecting coagulation factorⅧactivity was evaluated,and compared with coagulation assay.The clinical application was evaluated by detecting FⅧactivity in acquired hemophilia A patients and patients with lupus anticoagulant by chromogenic substrate assay.Results The results of two constant quality control products were within range provided by the manufacturer(the bias was 3.93%-6.79%).The within-run imprecision was 1.86%-2.06%(≤5%).The between-run imprecision was 4.83%-6.90%(≤15%).The chromogenic substrate assay had a good performance in sensitivity(CV=11.23%<20%).The recommended reference range was appropriate for our laboratory.The maximum dilution was 1∶16.The linear range was 5.00%-193.50%(a=1.0243,R2=1.000).The clinical reportable range was 0.50%-387.00%.The method had a low carryover rate(0.04%).The chromogenic substrate assay had good consistency with coagulation assay in detecting coagulation factorⅧactivity(R2=0.961)as well as the titer of FⅧinhibitor(R2=0.973).The difference of FⅧactivity in patients with lupus anticoagulant between these two assays was statistically significant(t=9.232,P<0.05).Conclusion The chromogenic substrate assay has a good performance in clinical detection of coagulation factorⅧactivity with wider clinical application and less interference.

关 键 词：凝血因子Ⅷ 发色底物法 临床应用评价 

分 类 号：R554.1[医药卫生—血液循环系统疾病]