一种高灵敏度的脂蛋白相关磷脂酶A2活性检测方法  

A High Sensitivity Method for the Detection of Lp-PLA2 Activity

作　　者：冯杰[1] 梁辰[1] 薛辉[1] 陆丽华 何秀丽 虞留明 高艳红[1] 颜光涛[1] FENG Jie;LIANG Chen;XUE Hui;LU Lihua;HE Xiuli;YU Liuming;GAO Yanhong;YAN Guangtao(Department of Clinical Laboratory center, The First Medical Center of the PLA General Hospital, Beijing 100853, China;Suzhou Evermed Biomedical Co., Ltd,Suzhou 215163, China)

机构地区：[1]解放军总医院第一医学中心医学检验中心,北京100853 [2]苏州博源医疗科技有限公司,江苏苏州215163

出　　处：《标记免疫分析与临床》2020年第5期850-855,共6页Labeled Immunoassays and Clinical Medicine

基　　金：国家重点研发计划课题子课题(编号:2017YFF0205401);国家科技支撑计划项目(编号:2015BAK45B01)。

摘　　要：目的酶法测定脂蛋白相关磷脂酶A2(Lp-PLA2)活性。方法脂蛋白相关磷脂酶A2水解底物1-豆寇酰基-2-(4-对硝基苯酚丁二酸酐)磷脂酰胆碱Sn-2位置,生成一种有颜色的产物4-对硝基苯酚,该产物在405 nm处有吸收峰,根据其吸光度的变化速率计算Lp-PLA2的活性。结果据本研究建立的方法,健康人参考范围0~670 U/L,线性范围为42.0~1658.0U/L,准确度相对偏差≤10%,批内变异<5%,批间变异<10%,携带污染率和试剂上机21d稳定性等符合判断标准。与德国Desai公司Lp-PLA2试剂检测结果进行线性回归分析,决定系数R2=0.991(n=20),一致性较好。结论该Lp-PLA2酶法活性测定的试剂盒性能满足临床检测要求,可以推广使用。Objective To determine the activity of Lp-PLA2 by an enzymatic method.Methods A colored product,4-p-nitrophenol,was produced from the hydrolysis of substrate 1-myristoyl-2-(4-nitrophenylsuccinyl)phosphatidyl choline sn-2 by lipoprotein phospholipase A2.The product had an absorption peak at 405 nm.The activity of Lp-PLA2 was then calculated according to the change rate of absorbance.Results The method established in this study had the linear range of 42.0-1658.0 U/L and the reference range for healthy people was 0-670 U/L.The relative accuracy deviation was less than 10%,the intra assay variation was less than 5%,the inter assay variation was less than 10%,the carrier contamination rates and the 21 days stability of the reagent on the Biochemical analyzer met the criteria.The correlation coefficient(R2=0.991,n=20)was in a good agreement with the results of Lp-PLA2 reagent of Desai company.Conclusion The performance verification of the Lp-PLA2 enzyme activity determination kit meets the requirements of clinical testing and can be popularized for the clinical practice.

关 键 词：脂蛋白相关磷脂酶A2 酶活性法 连续监测法 

分 类 号：R446.1[医药卫生—诊断学]