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

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

http://www.100md.com 《实用医技杂志》 2006年第23期

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,,],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,应用PVA构

     [摘要] 目的：合成(SA)z(Bio)xPVA(BCPDA)yEu3+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Eu3+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Eu3+复合物，构建PAPPA试剂检测灵敏度为0.7 mIU/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Eu3+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。

    [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

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

    CHEN Weibu,WU Xuecheng,GENG Weiming

    (The Laboratory Medicine Department of Shenzhen People's Hospital, Shenzhen， Guangdong 518000，China)

    Abstract :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Eu3+. Methods Polyvinylamine (PVA) was labeled with biotinstreptavidin and europium chelate of 4,7bis(chlorosulfophenyl)1,10phenanthroline2,9dicarboxylicacid(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 conjuagete of (SA)z(Bio)xPVA(BCPDA)yEu3+ and a reagent of PAPPA. Detection limits achieved were 0.7mIU/L.The calibration curve was linear 010000mIU/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Eu3+ 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. ......

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