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1、临床医学论文•用液相芯片技术定量测定人血清CEA・AFP和HBsAg作者:陈玮,李明,吴英松,李妙艳,何蕴韶【摘要】【目的】采用液相芯片技术同时定量测定人血清中癌胚抗原(CEA)、甲胎蛋白(AFP)和乙肝表面抗原(HBsAg),并对该方法进行评价。【方法】制备抗体交联微球及生物素标记抗体,用双抗体夹心法对临床血清标本进行测定。【结果】同时检测CEA、AFP和HBsAg时的线性范围分别为0.032〜200ng/mL'0.022〜55U/mL、0・26〜1000ng/mL,最低检测限分别为8.90pg/mL、0.013U/mL'0.24ng/mL,批内变异系数(CV)<9%»批间CV<14%。
2、检测CEA、AFPtiBsAg的灵敏度分别为96.4%95.8%92%特异度分别为95.6%94.2%、100%,准确度分别为96%、95%、96%。其检测结果与化学发光免疫分析法(CLTA)或时间分辨荧光免疫分析法(TRFIA)之间呈显著的等级相关关系。该方法仅需1?滋L标本,3h即可完成检测。【结论】液相芯片技术具有可联合检测多项指标、高通量、线性范围广、灵敏度高、重复性好、节省样品和时间等优点,具有临床应用潜力。【关键词】液相芯片;微球悬浮阵列技术;Luminex:肿瘤标志物;乙型肝炎表面抗原Abstract:[Objcctivc】Tosimultaneouslyquantifyca
3、rcinocmbryonicantigen(CEA),alphafetoprotein(AFP),andhepatiIisBsurfaceantigen(HBsAg)inhumanscrumusingmultiplexedLumincxassayandevaluatetheassayperformance.【Methods】Biotinylateddetectionantibodiesandmicrospherescoupledwithcaptureantibodieswereprepared.Asandwichimmunoassaywascstablishedtomeasuretheth
4、reeanalytesinserumsamples・【RcsulIs】Linearrangeswere0.032〜200ng/mL,0.022〜55U/mL,0.26〜1000ng/mL,anddetectionlimitswere8.90pg/mL,0.013U/mL,0.24ng/mLforCEA,AFP,andHBsAg,respectively.Theintra-andinter・dssnycoefficientsofvariationwerelessthan9%and14%respectivcly.FotCEA,AFP,andHBsAg,sensitivitywere96.4%,
5、95.8%and92%,specificitywere95.6%,94.2%and100%,andaccuracywere96%,95%and96%,respectively.SignificantSpearmanrankcorrelationswerefoundbetweentheresultsobtainedbyLuminexassayandbychemiluminescenceimmunoassay(CLIA)ortime-resolvedfluorescenceimmunoassay(TRFIA).Only1microliterofscrumand3hourswereneededf
6、orLuminexassay.【Conclusion】Withtheadvantagesofuniquecapabi1ityofmultiplexingandhigh-throughput,goodreproducibi1ity,increaseddetectionrangeandsensitivity,Luminexassaywouldhaveagreatpotentialinclinicalapplication.Keywords:1iquichip;suspensionbeadarray;Luminex;tumormarkers;hepatiIisBsurfaceantigen肿瘤[
7、JSUNYat-senUniv(McdSci),2007,28(1):105-109;114]标志物对于肿瘤普查、诊断、分期、疗效监测及预后判断等具冇重耍价値,临床上常联合检测多种标志物以提高肿瘤诊断的阳性率和特异性,但目前常用的检测手段如放射免疫测定、酶联免疫测定、化学发光法等仅能单独检测。液相芯片是一种新型生物芯片技术平台,它在不同荧光编码的微球上进行抗原■抗体及核酸杂交反应,通过两束激光分别检测微球编码和荧光信号以