25-羟基维生素D化学发光定量免疫分析方法的建立  

The Establishment of Chemiluminescence Quantitative Immunoassay for 25-hydroxyvitamin D

作　　者：周齐洋 徐海伟 梁辰[3] 冯杰[3] 颜光涛[3] ZHOU Qiyang;XU Haiwei;LIANG Chen;FENG Jie;XUE Hui;YAN Guangtao(Jiangsu Institute of Medical Device Testing,Nanjing 210012,China;SuzhouHybiome Biomedical Engineering Co.,LTD,Suzhou 215163,China;Center for Clinical Laboratory Medicine,PLA General Hospital,Beijing 100853,China)

机构地区：[1]江苏省医疗器械检验所,江苏南京210012 [2]苏州长光华医生物医学工程有限公司,江苏苏州215163 [3]中国人民解放军总医院医学检验中心,北京100853

出　　处：《标记免疫分析与临床》2021年第1期127-131,169,共6页Labeled Immunoassays and Clinical Medicine

基　　金：国家科技支撑计划(编号:2015BAK45B01)。

摘　　要：目的建立一种竞争法定量检测人血清血浆中25-羟基维生素D[25-(OH)D]的免疫分析方法。方法本研究以磁颗粒-链酶亲和素-生物素为固相分离系统,样本中加入解离剂使维生素D结合蛋白(vitamin D binding protein,VDBP)变性而使25-(OH)D游离出来,用生物素标记25-(OH)D,用吖碇酯标记一株抗-25-(OH)D的绵羊单抗,溯源到国际标准品,建立25-(OH)D定量免疫分析方法。结果此分析方法空白限不高于1.0 ng/mL;批内不精密度不超过5%,批间不精密度不超过10%;在线性范围为6~200 ng/mL;与金标准方法LC-MS/MS相比,相关性在0.97以上,偏差在±30%内,无系统偏差。结论该25-(OH)D化学发光定量检测方法,性能优异,适合临床推广使用。Objective To establish a competitive immunoassay for the quantitative determination of 25-hydroxyvitamin D[25-(OH)D]in human serum and plasma samples.Methods In this study,a magnetic particle-streptavidin-biotin solid phase separation system was used.Vitamin D Binding Protein(VDBP)was denatured and 25-(OH)D was freed by dissociator.25-(OH)D was labeled with biotin and an anti-25-(OH)D sheep monoclonal antibody was labeled with acridine ester.We then traced back to the international standard and established 25-(OH)D quantitative immunoassay method.Results The LOB of this method was less than 1.0 ng/mL.The intra-batch and inter-batch precision were less than 5%and 10%.The linear range was 6-200 ng/mL.Compared with the gold standard LC-MS/MS,the correlation was over 0.97,the deviation was less than 30%,and there was no systematic deviation.Conclusion This 25-(OH)D chemiluminescence quantitative detection method has an excellent performance and is suitable for clinical applications.

关 键 词：25-羟基维生素D LC-MS/MS 吖碇酯化学发光 24 25-二羟基维生素D 

分 类 号：R446.61[医药卫生—诊断学]