妊娠相关血浆蛋白A 定量测定方法的建立.PDF

356 栽蚤葬灶躁蚤灶 酝藻凿 允熏 May 圆园10熏 灾燥造 38 晕燥 5 妊娠相关血浆蛋白A 定量测定方法的建立* 李灵俊 陈淑莲 李向东 彭 静 李会强 王 谦吟 摘要 目的:建立人外周血妊娠相关血浆蛋白A（PAPP-A）定量酶联免疫测定方法。方法:以生物素化牛血清白蛋 白-链霉亲和素间接包被微孔板，将标记有生物素的抗 PAPP-A 抗体作为捕获抗体，结合标本中的抗原成分；标记有 辣根过氧化物酶的抗 PAPP-A 抗体作为检测抗体，经酶催化底物显色，定量测定人体外周血PAPP-A。结果:所建立的 测定方法灵敏度为0.31 mg/L ，回收率均在95%~100%之间；该方法线性、稳定性及特异性良好；3 份低、中、高浓度的血 清标本重复测定 10 次，批内变异系数（CV ）均小于10% ，批间CV 均小于15% ；该方法测量结果与国外同类方法测量 结果具有相关性（r 越 园援怨愿 ，孕约 园援05 ），2 种检测方法高度相关。结论:本文采用间接包被技术所建立的人外周血PAPP- A 固相酶联免疫测定方法灵敏、可靠、稳定性好，与国外同类方法的试剂盒之间相关性较高。 关键词 妊娠相关血浆蛋白A 间接包被 酶联免疫吸附测定 血清白蛋白, 牛 Establishment of Quantitative Determination Method for Pregnancy-Associated Plasma Protein A LI Lingj un, CHEN Shulian, LI Xiangdong, PENG Xueji ng, LI Huiqiang, WANG Xueqian Tianj in Medical Univ ers ity, Tianj in 300070, China Abstract Objective: To establish an enzyme immunoassay for detecting the concentration of pregnancy -associated plasma protein A (PAPP-A). Methods: The plates were coated with biotinylation bovin serum albumin and avidin-enzyme- linked. A biotin-labeled antibody for PAPP-A was used as catching antibody to capture the antigen of PAPP-A in the sample. The another antibody for PAPP-A that labeled with horseradish peroxidase was used as detecting antibody to test the concen鄄 tration of PAPP-A in human serum. Results: The sensitivity of the assay we developed was 0.31 mg/L and its recovery rates were between 95% and 100%. The linearity, stability and specificity of the assay were good. The three serum samples that con鄄 tained PAPP-A each at low, middle and high levels were tested repeatedly 10 times, showing that the coefficient of variations (CVs) were not exceed 10% in the group, and the CVs were not exceed 15% between groups. The assay we