豬瘟膠體金免疫層析快速診斷方法的建立.pdf

1、河北農(nóng)業(yè)大學碩士學位論文豬瘟膠體金免疫層析快速診斷方法的建立姓名：陳龍賓申請學位級別：碩士專業(yè)：預防獸醫(yī)學指導教師：孫繼國李佩國20050617RapidDetectionofClassicSwineFeverVirusbyColloidalGoldConjugatedAntibodyTestPaperAuthor：Specialty：Supervisor：ChenLong—binPreventiveVeterinaryMedicine

2、ProfSunJi—guoAbstractAsimp。lemethodwasdevelopedforthedetectionofclassicswinefevervirusinclinicalsamplesusinggoldimmunochromatography(GICA)11kclassicswinefevervirusvaccinewasusedtoimmunizerabbitsandcocksTheimmunizedserawe

3、repurifiedbycaprylicacid／ammoniumsulfamateprecipitationreactionTheresultantinununoglobulinG(IgGlwasanalyzedbytheconventionalPAGEandagardiffusionassayThePAGEshowedthatthepurifiedantibodiesconsistofYglobulinandalittleother

4、globulinTheactivationoftwokindsofimmunizedseraandpurifiedIgGinagardiffusionassaywerel：16Tri—sodiumcitratewithaqueousgoldchloridewerewarmedupandmixedtosynthesizegolds01The15umcolloidalgoldparticleswereselectedfromtheprima

5、rypreparationofcolloidalgoldnanoparticleswhichshoweddifferentcolorandjudgedesailyThemulticlonalantibodyagainstclassicswinefeverviruswaslabeledwiththeselectedgolds01Whenlmilliliteroftheselectedgoldsolwaslabeled，40txgpurfi

6、edantibodieswereneededatleastThelabeledrabbitlgGagainstclassicswinefeverviruswaspurifiedonlowtemperatureultracentrifugingTheIabeledrabbitIgGagainstclassicswinefeverviruswasdilutedinTBS(containingl％BSAlattheratioofl：2Thec

7、hickenanti—CSFVantibodyandsheepanti—rabbitantibodywereimmobilizedtodifferentareasofnitrocellulosemembraneTheoptimizedconcentrationsoftwoantibodieswere7509晷／mLand50099／mLrespectively，intermsoftitrationofthesquarematrixtes

8、tAgoodrecipewasvalidandwasusedtopreparethestoringlabeledrabbitIgGagainstclassicswinefevervirusOurresultshowedthepotentialofdetectingandreleasingWasverypromisingwhenthetestpaperwasstoredat4“Cindesiccatorfor4monthesTheclin

9、icalsamplesofclassicswinefeverweredetectedbythetestpaperAtthesanletimetwodifferentmethodsincludingFAandRTPCRweretestedinparallelThcresultshowedthatthismethodwaseasytobejustified，timesaving，differentialandsuitableforfield