四環(huán)素多克隆抗體的制備及丁胺卡那霉素酶聯免疫檢測的初步研究.pdf

1、鄭州大學碩士學位論文四環(huán)素多克隆抗體的制備及丁胺卡那霉素酶聯免疫檢測的初步研究姓名：俎晶晶申請學位級別：碩士專業(yè)：藥物分析化學指導教師：屈凌波李建軍20070501鄭J，I大掌qE掌位能。文AbstractTetracyclinefrc)isasyntheticantibacterialtthatbelongstotheTetracyclinesgroupandAmikacin(Amk)isoneofthemostCOlnmOllsem

2、isynthespenicillinthatbelongstotheAminoglyensidegroupTheyhavebeenincreasinglyusedinveterinarymedicinetotreatmicrobialinfectiousHowevertheTCandAmkresiduesinanimaledibletissuescouldc捆lseseriouspublichealthproblemsEnzymelin

3、kedimmunosorbentassay(ELISA)isaconvenientandfastmethodtoscreenlargesamplesButnowmainlykitsdependonforeignsoitisurgenttodevelopELISAfastkitforclenbuterolwithowuindependentpropertyrightItsverymeaningfultoestabfishafastdete

4、ctionmethodtodetectthetwoantibioticsresiduetocontrolanimalfoodsafetyandsafeguardconsumers’healthThepurpo∞ofthestudyhereistodevelopanindirectcompetitiveenrynlelinkedimmunoanrbentassays(CiELISAltomonitorTCandAmkresiduesina

5、nimalfoodThemaincontentsandresultsweleasfollowed：1SynthesisandIdentificationofTetracyclineandAmikacinArtificialAntigenTheimmtmogenandthecoalingantigenweresynthesmedbycarbodimidemethodThecompleteantigenswereidentifiedbyUV

6、spectrometricscanningmethodandimmuneBALB／cmicetestsTheresultsdemonstratedthattheartificialantigensweresynthesizedsuccessfully2SynthesisandIdentificationofTetracyclineandAmikacinpolyclonalantlbodyTheantiserumtitrcandspeci

7、ficityobtainedusinganindirectELISAandCiELISArespectivelyTheresultsshowed：theantiserumtitreofTCwasabout1：80000andthespecificityWasgood；theantiserumtitreofAmikacinwasabout1：600000andthespecificitywasgood3TheCi—ELISAmethodw

8、asestablishedtodetectAmikaeinresiduesTheoptimaltestconditionswasasfollowed：coatingantigen，blockingreagent，andhorseradishperoxidasemarkedtothegoatIgGofantirabbitlgGweTe1：10001％ofseparatedmilkend1：5000respectivelyItsstanda

9、rdcluNeisin005rtg／mLlI|tg／mLthecurvyequationisy=04641x06958，coefficientcorrelationR_09955，塒血alimitofdetectionto40ng／raLIC，尹038pg／mLPrecisionoftheCiELISAindicatedbyeoeffcientsofvariationofintraassayandinterassayw∽197％、127

10、％、45％and201％、95％、8o％TherewasllocrossactivitybetweenthesanlekindandfiveotherkindsAccuracyofthemethoddenotedbymeanrecoveriesofmilkandkidneysamplewere81％、78％、75％and79％、76％、72％4SynthesisandIdentificationofAkacinmonoclonalant

11、ibodywereexpotmdedintheresearchTheCiELISAmethodwasestablishedtodetectAmikaeinresiduesThecurvyequationisy=04942x07554，coefficientcorrelationR=O9956，謝thalimitofdetectionto44ng／mL1C如o35pg／mLTherewasDocrossactivitybenv嘲thesa