慶大霉素免疫分析新方法的研究.pdf

1、江南大學碩士學位論文慶大霉素免疫分析新方法的研究姓名：馬偉申請學位級別：碩士專業(yè)：動物營養(yǎng)與飼料科學指導教師：胥傳來20090601AbstractAbstractGentamicinlSanaminoglycosideantibioticwhichwaswidelyusedintreatingbacterialinfectionsinanimalsbecauseofitsgoodspectrumofactivityagainstgra

2、m—positiveandgramnegativebacteriaThepresenceofgentamicinresiduesinfoodisconsideredasahighrisktoconsumerTheobjectiveofourresearchistodeveloparapidandreliableanalyticalmethodwhichiSusedtomonitorgentamicinresidueinanimaltis

3、suesTheimmunogenandthecoatingantigenweresynthesizedbyusinggentamicincoupledtocarrierproteinbovineserumalbumin(BSA)andovalbumin(OVA)respectivelyAfterthepolyclonalantibodiesagainstgentamicinwereobtained，anELISAmethodforthe

4、determinationofGentamicinwasdevelopedbyusingthepolyclonalantibodiesTheantiserumagainstgentamicinwasacquiredafterimmunizationinrabbitsThetiterofantiserumwas512000Todevelopgentamicinimmunoassaytheinfluenceofseveralfactorss

5、uchascoatingtime，competitiontime，andpHwerestudiedUnderoptimizedconditions，thedetectionlimitwas01ng／m1TheresultsindicatedthattheoptimalworkingdilutionofcoatingantigenandantibodyWas1：3200and1：16000Thebestconcentrtionofenzy

6、melabledsecondaryantibodywas1：4000TotestthespecialityofantibodythreeaminoglycosidedrugswereusedinthecompetitionELISAasthecompetitionantigenTheresultsindicatedthattheantibodynocrossreactionwithotheraminoglycodidesByusingt

7、heantibodyWeestablihedimmunochromatographicstripforgentamicinThedetectionlimitofthisassayis50ng／ml，whichissensitiveenoughtoscreengentamicinresidualAcordingtorealsampleanalysis，theresultagreeswellwithEnzyme—linkedimmunoso

8、rbentassay(ELISA)WealsosynthesizedmagneticresonanceimagingimmuinosensorforthedetectionofgentamicinWesuccessfullyestablishedthestandardcurveofgentamicinwithaequations：y21775x一244ThedetectionrangeforgentamicinisO91—24ng／m1