应用PVA构建妊娠相关血浆蛋白A固相时间分辨荧光免疫试剂  

Develop A Direct Solid Phase Lanthane Time-resolved Fluoroimmunoassay Reagent of PAPP-Autilizing PVA

作　　者：陈卫布 武学成 耿伟明 

机构地区：[1]深圳市人民医院,广东深圳518000

出　　处：《实用医技杂志》2006年第23期4113-4117,共5页Journal of Practical Medical Techniques

摘　　要：目的：合成（SA）z-（Bio）x-PVA-（BCPDA）y-Eu^3＋复合物，并用于构建妊娠相关血浆蛋白A固相时间分辨免疫荧光试剂。方法：用PVA作为载体结合BCPDA-Eu^3＋和生物素-亲和素，以双抗体夹心法原理，结合生物素标记抗体和包被抗体建立PAPP-A时间分辨免疫荧光试剂，并进行方法学评价。结果：成功的合成了活性的（SA）z-（Bio）x-PVA-（BCPDA）y-Eu^3＋复合物，构建PAPP—A试剂检测灵敏度为0．7mIU／L；批内变异系数在3．89％～4．96％之间，批间变异系数在7．35％-9．27％之间。结论：合成的（SA）z-（Bio）x-PVA-（BCPDA）y-Eu^3＋复合物较高的反应活性，可以用于多种试剂的构建。构建的PAPP—A试剂灵敏度高，具有潜在的临床应用价值。Objective To develop a direct solid phase lanthane time-resolved fluoroimmunoassayreagent of PAPP-A by synthesizing a complex of （SA）z- （Bio）x-PVA- （BCPDA） y-Eu^3＋. Methods Polyvinylamine （PVA） was labeled with biotinstreptavidin and europium chelate of 4,7-bis（chlorosulfophenyl）-1,10-phenanthroline-2,9-dicarboxylic-aeid（BCPDA）. Using labeled PVA complex, We designed two-site sandwich time-resolved fluoroimmunoassay of PAPP-A and evaluated assay characteristics of PAPP-A kit. ReSults We successed synthesizing a conjuageto of （SA） z： （Bio） x-PVA- （BCPDA） y-Eu^3＋ and a reagent of PAPP-A. Detection limits achieved were 0.7mIU/L. The calibration curve was linear 0-10000mlU/L. Intra- and interassay imprecision （CV） was 3.89% -4.96% and 7.35% -9.27%. Recovery was 95.8% -104.2%. Conclusions The conjugate of （SA）z-（ Bio）x-PVA-（BCPDA） y-Eu^3＋ synthesizied was reactive ,highly fouorescent. A sorts of kit could be synthesized , taking advantage of it. The reagent of PAPP-A was potentially suited for use in clinical with highly analytical sensitivity.

关 键 词：妊娠相关血浆蛋白A 固相时间分辨荧光免疫 亲和素-生物素-聚乙烯胺-4 7二氯磺苯基-1 10啡罗啉-2 9二羧酸-铕复合物 唐氏综合征 

分 类 号：R714.1[医药卫生—妇产科学] R446.6[医药卫生—临床医学]