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1、自配试剂免疫透射比浊比率法测定血清CIC【摘要】目的:寻求一种全自动仪测循环免疫复合物(CIC)的新方法,以取代传统方法繁琐的手工劳动和略显粗糙的定量结果。方法:自配PEG和BBS试剂,以康宁560全自动生化仪结合AU560全自动生化仪设置程序最佳参数,免疫透射比浊终点法多点定标PEG和BBS,比率法计算CIC值,以百分回收试验和重复性试验确定其准确度和精密度,以对照试验与传统PEG沉淀法作对比分析。结果:仪测CIC最佳参数设置为:血清:试剂=1∶8,λ1340nm,λ2450nm,内源性读数时间
2、20Sec,总体反应时间288Sec,总反应体积为180μl,37℃。本方法与传统PEG沉淀法的回收率、日内和日间变异系数(CV%)分别为97%~110%、3.5%、4.3%和90%~122%、6.8%、9.7%。与传统PEG沉淀法相比,准确性差异无显著性(P>0.05),两者具有较好的相关性(r=0.852),精密度则比手工方法高。结论:自配试剂上机检测CIC,其操作简便易行,且定量相对准确,重现性好,线性范围宽,完全可以取代繁琐的传统操作。【关键词】免疫透射比浊法比率法循环免疫复合物 Ser
3、umCICDeterminationByImmunologicalTransmissionTurbidimetryRatiowithSelfmixReagents Abstract:ObjectiveTolookforanewfullautomaticinstrumentalmethodforCirculatingImmuneComplexes(CIC)determination,toreplacethetedioushandicraftlaboroftraditionalmethodand
4、gainmoreaccurateresults.MethodsTrytomixBBSandPEGreagentsself,establishthebestprocedureparameterswithKangning560combinewithAU560,immunologicaltransmissionturbidimetry4pointscalibrationthePEGandBBS,computetheCICvaluewithratiomethod,thenjudgeitsaccuracya
5、nddegreeofprecisionbypercentagerecoveryratiostudyandrepeatabilitystudy,makecontrastexperimenttocomparewithtraditionalPEGprecipitationmethod.ResultsThebestprocedureparametersofCICinstrumentalmethodwere:Serum:Reagents=1:8,λ1340nm,λ2450nm,endogenousreadt
6、ime20Sec,totalreactiontime288Sec,totalreactionvolume180ul,37℃.Percentagerecoveryratio,insidedayanddaytimepercentagevariationcoefficient(CV%)ofthismethodandtraditionalPEGprecipitationmethodrespectivelyfor97%~110%,3.5%,4.3%and90%~122%,6.8%,9.7%.Compared
7、withtraditionalPEGprecipitationmethod,ithasnodifferenceinaccuracy(P>0.05),thetwohavegoodcorrelation(r=0.852),butdegreeofprecisionishigherthanhandicraftmethod.ConclusionSerumCICdeterminationbyimmunologicaltransmissionturbidimetryratiowithSelf3mixreage
8、nt,itsoperationissimpleandconvenient,alsoitshowsquantitativemeasurementrelativelyaccurate,goodrepeatability,widelinear,itmayreplacenittygrittytraditionalhandoperationcompletely.KeyWords:Immunologicaltransmissionturbidimetry;Ratio;Circulating