孕妇血清妊娠相关蛋白A鲁米诺氧通道免疫测定方法的建立  

Development of luminescent oxygen channeling immunoassay for pregnancy associated plasm a protein A

作　　者：郝颖新 侯丽英[2] 周宇捷[2] 张月香[1] 李会强[2] 

机构地区：[1]天津市中心妇产科医院检验科,天津300100 [2]天津医科大学医学检验学院临床免疫学教研室,天津300203

出　　处：《中国免疫学杂志》2014年第7期917-920,共4页Chinese Journal of Immunology

基　　金：国家“863”计划项目(2012AA022602);天津市滨海新区自主创新重大项目(2011-BK120017)

摘　　要：目的：建立孕妇外周血妊娠相关蛋白A的鲁米诺氧通道免疫测定方法。方法：采用1株鼠抗人PAPP-A单克隆抗体用于生物素标记，兔抗人PAPP-A多克隆抗体包被受体微球，与包被有链霉亲和素的供体微粒共同构成检测试剂，优化检测条件并评价检测性能。结果：本方法的批内及批间精密度为5.91％～7.94％和6.14％～9.69％；分析灵敏度2.8 mU／L，功能灵敏度4.6 mU／L，在2.8～8000 mU／L范围内线性良好；回收率为96.7％～100.3％，通过干扰性实验分析溶血、黄疸和脂血的干扰率均＜10％；当PAPP-A浓度高达8000 mU／L时，无Hook效应；本文所建立方法检测50份临床标本，结果与时间分辨荧光分析法的结果相关系数为0.989。结论：本研究建立的方法能够用于孕妇血清PAPP-A的定量测定，检测性能符合临床诊断试剂的要求。Objective：To develop a luminescent oxygen channeling immunoassay for pregnancy associated plasm a protein A.Methods：The monoclonal antibody of PAPP-A was labeled with biotin ,the polyclonal antibody of PAPP-A was coated on receptor particles.The LOCI reagents also contained sensitizer particles coated with streptavidin.The optimal test conditions and analytical performance of the method were studied.Results：The withinrun and the between-run coefficients of variation were 5.91%-7.94% and 6.14%-9.69%,respectively;the analytical sensitivity was 2.8 mU/L and the function sensitivity was 4.6 mU/L,good linear in 2.8 mU/L-8 000 mU/L range;the recovery rate was 96.7%-100.3%.The interference rate of hemolysis , icterus and triglycerides were less than 10%; there is no Hook effect of PAPP-A concentrations up to 8 000 mU/L;the correlation coefficient between clinical samples detection results and Time resolved fluoroimmunoassay analysis results was 0.974.Conclusion：This LOCI can be used for the quantitative of serum PAPP-A,and detection performance in line with the requirements of clinical diagnostic reagents .

关 键 词：妊娠相关蛋白A 鲁米诺氧通道免疫测定 均相发光免疫分析 定量分析 

分 类 号：R392-33[医药卫生—免疫学]