N末端脑利钠肽前体电化学发光检测方法的建立  

The Establishment of An Electrochemiluminescence Detection Method for N-Terminal Brain Natriuretic Peptide Precursors

作　　者：魏雪梅 梁辰[1] 薛辉[1] 冯杰[1] 邓文斌 周桂生 曾映 周明 颜光涛[1] WEI Xue-mei;LIANG Chen;XUE Hui;FENG Jie;DENG Wen-bin;ZHOU Gui-sheng;ZENG Ying;ZHOU Ming;YAN Guang-tao(Medical Laboratory Center of First Medical Center,PLA General Hospital,Beijing 100853,China;Shenzhen Pumen Technology Co.,Ltd.,Shenzhen 518055,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]深圳普门科技股份有限公司,广东深圳518055

出　　处：《标记免疫分析与临床》2019年第12期2130-2134,2155,共6页Labeled Immunoassays and Clinical Medicine

基　　金：广东省引进创新创业团队计划资助(编号:2014ZT05S003);深圳市海外高层次人才创新创业资金(编号:KQTD2014063010175739);国家科技支撑计划(编号:2015BAK45B01)

摘　　要：目的建立一种定量检测人血清中N末端脑利钠肽前体(NT-proBNP)的基于三联吡啶钌的电化学发光免疫分析方法。方法以双抗体夹心法为原理,以生物素和三联吡啶钌标记NT-proBNP的单克隆抗体,以链霉亲和素包被磁微粒,建立一种以磁颗粒-链霉亲和素-生物素系统实现固相和液相分离的电化学发光免疫分析方法(eCLIA)。结果本研究建立的分析方法,其最低检测限可达2.39 pg/mL,线性范围为5~35000 pg/mL,批内变异小于8%,批间变异小于8%,与罗氏公司NT-proBNP检测试剂盒相关系数r=0.9957,可报告范围为5~137844 pg/mL。试剂盒整体置于37℃7 d,各性能指标不变,试剂稳定性良好。试剂抗干扰性能良好。结论该方法具有灵敏度高、线性范围广、稳定性好、受环境影响小、检测结果准确等特点,各项指标均达到临床检测要求,适合临床推广使用。Objective Based on the principle of double antibody sandwich method,a method for the quantitative detection of N-Terminal pro-B-type natriuretic peptide(NT-proBNP)in human serum based on terpyridine pyridinium-based electrochemiluminescence immunoassay was established.Methods Following double antibody sandwich method,a monoclonal antibody against NT-proBNP was labeled with biotin and terpyridine,and magnetic particles were coated with streptavidin to establish a magnetic particle-streptavidin-Biotin system to achieve solid phase and liquid phase separation of electrochemiluminescence immunoassay(eCLIA).Results The analytical method established in this study had a minimum detection limit of 2.38pg/mL,a linear range of 5 to 35000pg/mL,also an intra-assay variation of less than 8%,and an inter-assay variation of less than 8%.When compared with a Roche NT-proBNP detection reagent,the box correlation coefficient was r=0.9957.Reportable range was 5-137844(pg/mL).After the whole reagent kit was placed at 37°C for 7 days,the performance indexes were unchanged and the reagent stability was good.In general,the reagent has a good anti-interference performance.Conclusion The developed method in the current study has the characteristics of high sensitivity,wide linear range,good stability,little environmental impact,and accurate detection results,etc.It meets the requirements of clinical testing and is suitable for clinical promotion.

关 键 词：N末端脑利钠肽前体 链霉亲和素-生物素 三联吡啶钌 电化学发光法 

分 类 号：R54[医药卫生—心血管疾病]