妊娠相关血浆蛋白a定量测定方法的建立

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1、356TianjinMedJ，May2010，Vol38No5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学静李会强王学谦摘要目的：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一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0．31ms／L，回收率均在95％～100％之间；该方法线性、稳定性及

2、特异性良好；3份低、中、高浓度的血清标本重复测定l0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。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EstablishmentofQuantitativeDeterminationMethodforPregnancy——Associa

3、tedPlasmaProteinALILingjun，CHENShu~an，LIXiangdong，PENGXuejing，LIHuiqiang，WANGXueqianTianjinMedicalUniversity,Tianjin300070,ChinaAbstractObjective：Toestablishallenzymeimmunoassayfordetectingtheconcentrationofpregnancy-associatedplasmaproteinA(PAPP-A)．Methods：n1eplateswere

4、coatedwithbiotinylationbovinserumalbuminandavidin—enzyme—linked．Abiotin-labeledantibodyforPAPP-AWasusedascatchingantibodytocapturetheantigenofPAPP-Ainthesample．n1eanotherantibodyforPAPP-Athatlabeledwithhorseradishperoxidasewasusedasdetectingantibodytotesttheconcen—tratio

5、nofPAPP-Ainhumanserum．Results：ThesensitivityoftheassaywedevelopedwasO．31ms／Landitsrecoveryrateswerebetween95％and100％．Thelinearity，stabilityandspecificityoftheassayweregood．Thethreeserultllsamplesthatcon-rainedPAPP-Aeachatlow，middleandhighlevelsweretestedrepeatedly10times

6、，showingthatthecoeflqcientofvariations(CVs)weI．enotexceed10％inthegroup，andtheCVswerenotexceed15％betweengroups．TheassaywedevelopedWasdonetheanalysisassociativelywithenzymeimmunoassaykitabroad，r=0．982(P<0．05)，whichindicatedthattlletwoassayswerecorrelatedgreatly．Conclusion：

7、Usingindirect-coatingtechnology．theassayweestablishedtodeterminePAPP—Aissensitiveandaccurate，andthestabilityisgood．Thereisgreatlycorrelationbetweentheassayswedevelopedandtheothersabroad．Keywordspregnancy——associatedplasmaproteinAindirect-coatingtechnologyenzyme-linkedimm

8、unosorbentas-sayselllmalbumin，bovine妊娠相关血浆蛋白A(pregnancy—associated标抗体的复合物，建立外周血PAPP—A定量酶联免plasmaprotein