血清孕酮同位素稀释液相色谱串联质谱候选参考方法的性能评价及临床应用  

Performance evaluation of serum progesterone measurement by ID-LC/MS/MS candidate reference methods and their clinical application value

作　　者：欧阳芬 张乔轩 严君 韩丽乔 王建兵 柯培锋 庄俊华 黄宪章 Ouyang Fen;Zhang Qiaoxuan;Yan Jun;Han Liqiao;Wang Jianbing;Ke Peifeng;Zhuang Junhua;Huang Xianzhang(Department of Laboratory Medicine,the Second Affiliated Hospital of Guangzhou University of Chinese Medicine(Guangdong Provincial Hospital of Chinese Medicine),Guangzhou 510120,China)

机构地区：[1]广州中医药大学第二附属医院(广东省中医院)检验医学部,广州510120

出　　处：《中华检验医学杂志》2022年第5期456-462,共7页Chinese Journal of Laboratory Medicine

基　　金：广州市高校创新创业(就业)教育项目(平台建设重点项目)(2020PT102);广州市科技计划项目(202002020038)。

摘　　要：目的建立本实验室血清孕酮同位素稀释液相色谱串联质谱(ID-LC-MS/MS)候选参考方法,并将其用于5种临床常规检测系统的性能评价,探讨不同检测系统检测血清孕酮的可比性。方法采用液液萃取的方法进行样本前处理,在正离子质谱模式下的反相液相分离检测人血清中孕酮,通过梯度洗脱,使单个样本检测时间控制在5 min内。为提高方法准确度,使用包括法建立标准曲线,根据美国临床和实验室标准化协会C62-A、EP15-A2、EP6-A2和EP9-A3等文件分别评价包括法和经典校准曲线法检测的灵敏度、正确度、精密度和特异度等性能,并评价5个血清孕酮临床常规检测系统的检测性能,与ID-LC-MS/MS法比较,评估其在医学决定水平2和25 ng/ml处的偏倚是否均<1/2允许总误差(TEa,12.5%)。结果本实验室血清孕酮ID-LC-MS/MS候选参考方法检测限为0.005 ng/ml,包括法和经典校准曲线法回收实验的回收率分别为97.95%~101.58%和96.88%~110.70%,包括法对认证标准物质的测量结果在其声明的不确定度范围内;包括法批内、批间变异系数(CV)均<3.0%,优于经典校准曲线法(2.48%~9.33%)。5种临床常规检测系统的精密度和线性范围均能达到要求;检测系统1、3和5在2和25 ng/ml处2个医学决定水平的测量结果偏倚不满足均<1/2TEa,检测系统2和4在2个医学决定水平的测量结果偏倚均<1/2TEa。结论本室建立的血清孕酮ID-LC-MS/MS候选参考方法分析性能符合要求,包括法较经典校准曲线法具有更好的检测性能;5种临床常规检测系统的精密度、线性评估良好,仅2个检测系统在2个医学决定水平的正确度评价能满足临床需求。Objective To establish a candidate reference method for serum progesterone using isotope dilution liquid chromatography tandem mass spectrometry(ID-LC/MS/MS)in our laboratory,validate the analytic performance of five clinical routine detection systems to explore the comparability of serum progesterone detection by different detection systems.Methods A candidate reference method for serum progesterone using ID-LC/MS/MS method was established.The sample was pretreated by liquid-liquid extraction method,and the reversed phase liquid phase separation in positive ion mass spectrometry mode was used to detect progesterone in human serum,and the detection time of a single sample was controlled within 5 minutes by gradient elution.In order to improve the accuracy of the method,the bracketing calibration method(BCM)was used to establish the standard curve.The sensitivity,accuracy,precision and specificity of BCM and classical calibration curve method were evaluated according to CLSI C62-A,EP15-A2,EP6-A2 and EP9-A3,and the analytical performance and comparability of five clinical routine progesterone detection systems were evaluated,compared with ID-LC/MS/MS method,the bias at medical decision level 2 and 25 ng/ml was evaluated to see if they were<1/2TEa(12.5%).Results The limit of detection(LOD)of ID-LC/MS/MS was 0.005 ng/ml.The recoveries of BCM method and classical calibration curve method are 97.95%-101.58%and 96.88%-110.70%,respectively.The measurement results of BCM method for certified reference materials are within its declared uncertainty range.The intra-and inter-assay coefficient of variation(CV)of BCM method was less than 3.0%,which was better than that of classical calibration curve method(CV:2.48%-9.33%).The precision and linear range of the five clinical routine detection systems can meet the detection requirements.The measurement bias of detection system 1,3 and 5 at 25 ng/ml of medical decision level was less than 1/2TEa,and the measurement bias at 2 ng/ml of medical decision level was more than 1/2TEa.The

关 键 词：孕酮 液相色谱串联质谱 参考方法 性能评价 

分 类 号：R446.1[医药卫生—诊断学]