人骨钙素检测化学发光法的建立及性能评价  

Establishment and evaluation of chemiluminescent assay for human osteocalcin

作　　者：许亚丰[1] 黄建荣[1] 薛一峰[1] 朱旭明 韩霜 王春新[1] XU Yafeng;HUANG Jianrong;XUE Yifeng;ZHU Xuming;HAN Shuang;WANG Chunxin(Department of Laboratory Medicine, Wuxi People's Hospital,Wuxi 214000,Jiangsu;School of Bioengineering,Jiangnan University,Wuxi 214122,Jiangsu ,China)

机构地区：[1]无锡市人民医院医学检验科,江苏无锡214000 [2]江南大学生物工程学院,江苏无锡214122

出　　处：《临床检验杂志》2018年第12期895-898,共4页Chinese Journal of Clinical Laboratory Science

基　　金：国家高技术研究发展计划(2015AA020802)

摘　　要：目的建立一种灵敏的定量检测人骨钙素的方法。方法采用磁微粒化学发光法定量检测人血清中的骨钙素含量,反应模式为双抗夹心法,生物素化捕获抗体,辣根过氧化物酶标记检测抗体与血清(校准品)及包被有链霉亲合素的磁微粒构成完整的分析系统。并对本方法进行条件优化、性能评估以及相关性分析。结果建立的人骨钙素磁微粒化学发光法线性范围为0.46～280.25 ng/mL,空白限为0.23 ng/mL,重复性和期间精密度(CV)分别小于8%和10%,回收试验回收率在103%～108%之间;血红蛋白、总胆红素和三酰甘油对骨钙素检测无干扰作用;在120例临床血清样本中,该方法检测骨钙素结果和罗氏电化学发光法结果相关系数(r)为0.992。结论建立的定量检测人骨钙素的磁微粒化学发光法精密度好、线性范围宽、灵敏度高,达到临床测定的需求。Objective To establish a sensitive method for quantitatively determining human osteocalcin. Methods The established chemiluminescent immunoassay(CLIA)using magnetic particles for determining osteocalcin was performed by incubating serum samples or calibrator with biotinylated capture antibody,horseradish peroxidase(HRP)labeled detecting antibody and streptavidin coated mag netic particle. The working conditions of this assay were optimized. The performance was evaluated and the correlation of osteocalcin re sults between CLIA and other system was analyzed. Results The CLIA assay of osteocalcin was successfully established. The linear range was 0.46 to 280.25 ng/mL. The limit of blank was 0.23 ng/mL. The repeatability and intermediate precision were less than 8% and 10% respectively. The recovery rates measured by adding osteocalcin calibrators to human sera with different osteocalcin concentra tions were ranged from 103% to 108%. No interference was observed by adding hemoglobin,total bilirubin and triacylglycerol to human sera with different osteocalcin concentrations. The correlation coefficient of osteocalcin results between this CLIA method and Roche electrochemiluminescence immunoassay system was 0.992. Conclusion The quantitative detection for osteocalcin by CLIA established in this study showed better precision,wide linear range and high sensitivity,so as to meet the requirements of clinical measurement.

关 键 词：磁微粒 化学发光法 双抗夹心法 骨钙素 

分 类 号：R446.6[医药卫生—诊断学]