超敏妊娠相关血浆蛋白A均相化学发光免疫检测方法的建立  

Establishment of homogeneous chemiluminescent immunoassay method of pregnancy-associated plasma protein A

作　　者：王瑞[1,2] 曾昭伟[2] 李会强[1] 侯丽英[1] 王雪颖[1] 周玉章 

机构地区：[1]天津医科大学医学检验学院,天津300203 [2]天津市南开医院,天津300100

出　　处：《中国卫生检验杂志》2015年第4期507-510,共4页Chinese Journal of Health Laboratory Technology

基　　金：超敏PAPP-A均相发光免疫测定方法建立及初步应用基金(2013KY24);高通量均相发光免疫分析仪与诊断试剂开发基金(2012AA022602)

摘　　要：目的建立有关急性冠脉综合征(ACS)患者血清中妊娠相关血浆蛋白A(PAPP-A)的均相化学发光免疫检测方法。方法使用PAPP-A抗体与受体微球和生物素连接,以双抗体夹心法,生物素化抗体和受体微球与血清中PAPP-A结合,以鲁米诺氧途径免疫分析(LOCI)检测PAPP-A浓度。评价此方法各项性能指标。数据分析使用ELISA Calc软件。结果本研究PAPP-A检测方法灵敏度为0.55 m U/L,回收率为95%~100%;线性、稳定性及特异性良好;重复性批内变异系数(CV)<10%,批间CV<15%;检测不易受血红蛋白、甘油三酯和胆红素的干扰。该方法与时间分辨荧光免疫方法(TRFIA)进行相关分析(r=0.990,P<0.05)。检测104例正常健康人(男/女为43/61,年龄为24岁~69岁)血清PAPP-A,初步推算正常参考范围为1.60 m U/L^25.12 m U/L。结论本研究所建立的超敏PAPP-A检测方法准确性好,灵敏度、回收率、特异性较高,与TRFIA相关性良好,评价了其应用于临床检测的可行性,并为参考范围的确定提供了一定的依据。Objective To establish a homogeneous chemiluminescent immunoassay for detecting pregnancy- associated plasma protein A（ PAPP- A） in patients with acute coronary syndrome（ ACS）. Methods To connect PAPP- A antibody with receptor microspheres and biotin,with double antibody sandwich assay,to combine biotinylated antibody and PAPP- A receptor with microspheres in serum,by immune lumino oxygen pathway analysis（ LOCI） to detect the concentration of PAPP- A. Evaluation of the performance index of this method was conducted. The data were analyzed by using ELISA Calc software. Results The sensitivity of the assay we developed was 0. 55 m U / L and its recovery rates were between 95% and 100%. The linearity,stability and specificity of the assay were good. The repeatability showed that the coefficient of variations（ CVs） were less than 10%in the group,and the CVs were less than 15% between groups. The testing results were not subjected to interference by hemoglobin,triglycerides,or bilirubin. The analysis was done with the assay associatively with TRFIA（ r = 0. 990,P 0. 05）.PAPP- A of 104 healthy serum samples（ M / F： 43 /61,Age： 24- 69） were detected,the range were deduced between1. 60 m U / L- 25. 12 m U / L. Conclusion The assay we established to determine PAPP- A is sensitive and accurate,and the stability is excellent. There is greatly correlation between the assays we developed and the TRFIA. The posibity of applying in chinic was evaluated,and the basis of conforming reference range was provided.

关 键 词：均相免疫分析 鲁米诺氧途径免疫分析 急性冠脉综合征 妊娠相关血浆蛋白A 检测方法 

分 类 号：R446.6[医药卫生—诊断学]