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0.31mg/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=0.982,P〈0.05）,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。
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-enzymelinked.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 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 contained 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 developed was done the analysis associatively with enzyme immunoassay kit abroad,r = 0.982 （P 0.05）,which indicated that 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 is greatly correlation between the assays we developed and the others abroad.
出处
《天津医药》
CAS
北大核心
2010年第5期356-359,共4页
Tianjin Medical Journal
基金
天津市科技支撑计划项目(项目编号:08ZCGYSF01800)
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
pregnancy-associated plasma protein A indirect-coating technology enzyme-linked immunosorbent assay serum albumin
bovine
