天津医药

• 论著 •    

妊娠相关血浆蛋白A定量测定方法的建立

李灵俊1,陈淑莲2,李向东2,彭学静2,李会强2,王学谦3   

  1. 1. 天津医科大学
    2.
    3. 天津市天津医院
  • 收稿日期:2009-12-07 修回日期:2010-03-04 出版日期:2010-05-15 发布日期:2010-05-15
  • 通讯作者: 李灵俊

The studies on the method for the quantitation of pregnancy-associated plasma protein A

  • Received:2009-12-07 Revised:2010-03-04 Published:2010-05-15 Online:2010-05-15

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摘要： 摘要 目的:采用微孔板间接包被法，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-链酶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性为0.31μg/ml，回收率均在95%-100%之间；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CV均不大于10%，批间CV均不大于15%；与国外同类方法进行相关性分析，r=0.982，提示两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固相酶免疫测定方法灵敏、可靠、稳定性好，与国外同类试剂之间相关性较高。

关键词: 妊娠相关血浆蛋白A, 间接包被, 酶免测定

Abstract: Abstract Objective: To develop an Enzyme immunoassay for testing the concentration of pregnancy-associated plasma protein A, the indirect-coating plates were adopted. 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 concentration of pregnancy-associated plasma protein A in human serum. Results: The sensitivity of the assay we developed was 0.31μg/ml and its recovery rates all between 95% and 100%.The linearity, stability and specificity of the assay are good. The three serum samples that the concentration of PAPP-A each at low, middle and high levels were tested repeatedly 10 times, showing that the CVs inner the group all did not exceed 10% and the CVs between groups all did not exceed 15%.The assay we developed was done the analysis associativity with enzyme immunoassay kit abroad, r=0.982, which indicated the two assays were correlated greatly. Conclusion: Using indirect-coating technology, the assay we established to determine PAPP-A is sensitive and accurate, and the stability is good. There are greatly correlated between the assays, we developed and abroad.

Key words: pregnancy-associated plasma protein A, Indirect-coating technology, Enzyme immunoassay