人C肽时间分辨免疫荧光分析试剂盒的研制3

:10012764X(2005)0320200203:R587:AC31,2,2,2,1,1,1,2(11,510663;21,510515):C(C2peptide)(TRFIA)CTRFIA,0.522ng/ml,0.045ng/ml,(CV)4.1%6.7%5.4%6.8%;,9.0%30,0.992,:C;;C,(EIA)(RIA)(CLIA)(TRFIA)[1],,C2peptideTRFIA11.12CBiodesignMedixCSigma;96(812)Labsystem;SephadexG250Pharmacia;CEu3+PerkinElmer;WallacDELFIATMC2peptidekits,Auto2Delfia1235TRFIA30C430C1.250mmol/LpH9.63g/ml,96,200l,4,,(20g/LBSA),250l,372h,,,-201.3Eu3+0.5mgMillipore,8000r/min56min;(50mmol/L,Na2CO3,pH9.0)200l,8000r/min56min,6;,20003000r/min1min,,,0.05mg,251.4Eu3+SephadexG250(1cm30cm),(50mmol/lTris2HCl,9g/LNaCl),(1ml/),(A280nm),,,PerkinElmerEu3+Eu3+Eu3+(Eu3+mol/IgGmol)10.97,90%1.52g/LBSA,1g/LNaN3,50mmol/LPH7.8Tris2HClC0.5,4.7,13.7,22ng/ml,1ml,-201.625l,200l175200l,2h;6;200l5minAuto2Delfia1235Auto2Delfia1235TRFIA22.1,(log2log),2,0022005233ChineseJournalofClinicalLaboratoryScience,2005,Vol123,No133:()(2004Z12E4021):,1971,,,,E2mail:[e-mail],02023229078921640.471,0.466(tP0.05,)C,12r=0.99982.20.522ng/ml2.3(A)20€x2s,0.045ng/ml2.43(,2.06.512.0ng/ml),,8231C2TRFIA(n=8)ABCDE00.504.713.722.00(00.04)0.47(0.450.49)4.5(4.314.78)13.0(12.713.9)21.6(20.922.8)-0.94(0.900.98)0.96(0.911.02)0.95(0.931.01)0.98(0.951.04)2(€xs)040215C(ng/ml)CV%040612C(ng/ml)CV%040701C(ng/ml)CV%2.660.1084.082.480.1626.542.630.1455.517.480.4465.968.120.4215.198.260.5356.4813.790.8876.4314.550.9046.2114.950.9996.683(n=8)C(ng/ml)CV%2.620.1786.797.740.4265.5011.20.6105.392.5C(289IU/ml)(10ng/ml),01002mg/ml019ng/ml2.6430,222(1882),208(2173),03.75ng/ml,(€x)1.95ng/ml,(s)0.83ng/ml49717%0.33.6ng/ml,,0.33.6ng/ml4430C(ng/ml,n=8)00.50.61.01.11.51.62.02.12.52.63.03.13.53.5(%)3(0.69)24(5.58)76(17.67)236(54.88)63(14.65)18(4.19)8(1.86)2(0.47)2.7Wallac,,530,29,1;30,2,Wallac10.9925WallacWallac(ng/ml)0.0450.015(ng/ml)0.522100.42010CV(%)6.551041413CAB,31,3000,111C,,[5],,2,CC,,,[627]TRFIA,,,TRFIA,[4]EIARIACLIA,1022005233ChineseJournalofClinicalLaboratoryScience,2005,Vol123,No13[829],,,,25l,:[1],.[M].4.:,1997.7251[2],.C2[J].,2002,11(23):238722388.[3]PfutznerA,LobigM,FortunatoA,etal.Evaluationofanewfullyautomatedone2stepC2peptidechemiluminescenceassay(LIAISONC2Peptid)[J].ClinLab,2003,49(526):2272232.[4],1[J]1,2004,22(4):31323151[5].[J].,1999,7(1):57261.[6]BonserA,Garcia2WebbP.C2peptidemeasurement:methodsandclinicalutility[J].CRCCritRevClinLabSci,1984,19:2972352.[7]RendellM.ExpandingclinicaluseofC2Peptideradioimmunoassay[J].ActaDiabetolLat,1983,20:1052113.[8]YamadaH,ChibaH,AmanoM,etal.Rainbowtrouteyed2stageembryosdemonstratemelatoninrhytoninrhythmsunderlight2darkconditionsasmeasuredbynewlydevelopedtime2reslovedfluoroimmunoassay[J].GenCompEndocrinol,2002,125(1):41246.[9]AndohT,NagasawaH.Developmentofatime2resolvedfluoroimmunoassayforinsulinsanditsapplicationtomonitoringofinsulinsecretioninducedbyfeedinginthebarfinflounder,Veraspermoseri[J].GenCompEndocrinol,2002,125(3):3652374.Preparationoftime2resolvedfluoroimmunoassaykitfordetectionofhumanC2peptideTANGYong2ping1,WUYing2song2,HANGJian2feng2,XUWei2wen2,SUNSheng2li1,YUWei2hong1,ZHONGZhi2cong1,LIMing2(1.GuangzhouDaruiAntibodyCo.Ltd,Guangzhou510663;2.InstituteofTropicalMedicine,TheFirstMilitaryMedicalUniversity,Guangzhou510515)Abstract:ObjectiveToprepareatime2resolvedfluoroimmunoassay(TRFIA)kitfordetectionofhumanC2peptide.MethodsAsandwichTRFIAkitfordetectionofC2peptidewasdevelopedusingmonoclonalantibodytohumanC2peptide.ResultsThemeasurementrangeofthedevelopedkitwas0.5222ng/ml.Itssensitivitywas0.08ng/ml.Therewasnocross2reactionwithinsulinandthecrossreactivityratewithproinsulinwas9%.Theintra2andinter2assaycoefficientsofvariationwere4.126.5%and5.426.8%respectively.Thecorrelationcoefficientbetweenhome2madeandcommerciallyavailableC2peptidekit(PerKinElmer)fortheresultsofbloodsampleswas0.992.ConclusionThedevelopedC2peptideTRFIAkitinthisstudywasavailableforclinicalapplicationwithfinesensitivity,specificityandaccuracycorrespondtorelatedlaboratorycriteriaandmaysubstituteforforeignlikeproducts.Keywords:humanC2peptide,timed2resolvedfluoroimmunoassay(TRFIA),kit(:2004210220,:2005202220)(:)(1,[e-mail];2;[e-mail];3,[e-mail]),,(),(),772022005233ChineseJournalofClinicalLaboratoryScience,2005,Vol123,No13