金黃色葡萄球菌血清8型莢膜多糖的提取純化及其偶聯(lián)物免疫學特性的研究.pdf

1、分類號S852.3學校代碼10129UDC11.220學號2009201088慢性瘤胃酸中毒對瘤胃發(fā)酵功能及乳酸代謝菌的影響金黃色葡萄球菌血清8型莢膜多糖的提取純化及其偶聯(lián)物免疫學特性的研究StudytheextractionpurificationofCP8theimmunogenicityofconjugateinCP8fromStaphylococcusaureus申請人：王姝懿學科門類：農(nóng)學學科專業(yè)：預防獸醫(yī)學研究方向：獸醫(yī)微生

2、物與免疫學指導教師：郝永清教授論文提交日期：二〇一二年五月StudytheextractionpurificationofCP8theimmunogenicityofconjugateinCP8fromStaphylococcusaureusAbstractStaphylococcusaureusisoneofthemostsignificantpathogenswhichcausesthebovinemastitis.weconsid

3、erdthemajvirulencefactsiscapsularpolysacideadhesinofS.aureus.WhenS.aureusinvasionofthebodyitcan’tbeclearedeasilybyphagocyticcellbecauseofresistingthephagytosisofcapsularpolysacide.Furthermethebacterialadhesinrecognisedad

4、heredtotheganizationdefinitemoleculeofcell.Theinfectionishardtocontrolwhenbacteriumproduceenzymestoxinsafterthesuccessofcolonization.IntheexperimentCouplingthecapsulepolysacideadhesintopreparetheconjugatesthroughchemical

5、method.SoitlaidthetheeticalexperimentalbasisofthedivalentvirulencefactsvaccineofCPadhesininS.aureus.WeaddeddifferentnutrientsinColumbiasolidmediumindertooptimizethecultureconditionsofS.aureusinthetext.Usingthemethodofplu

6、senzymetoextractedCP8fromS.aureus.WepurifiedCPusingthroughcolumnchromatographymethods.ItscontentsweredetectedbythemethodofPhenolvitrioliccolimetry.OnthebasisofgetthepurifiedCP8wechoseADHasbridgeEDCasthecouplantthepurifie

7、dClfAFnBPBascarrierproteintobideCPClfAFnBPBtogetherindertoprepareconjugates.Inthecoupingprocesstodeterminethebestmassratioofconjugate.SecondlytheuseofUVabsbancePhenolvitrioliccolimetrymethodtoqualitativequantitativeanaly

8、siscalculatethecouplingratio.FinallytheBalBcmicewereimmunizedwiththeconjugatesELISAmethodmousetoxicitytestwerechosentodetectantibodylevelsprotectiveeffectofCPconjugates.TheresultshowedthatCP8S.aureushadthemostvigousgrowt

9、hinColumbiasolidmediumwhichadded1﹪lactose2﹪NaCl10﹪fetalbovineserum.Weusedtheseconditionstoculturedstrainswhichextractedcapsularpolysacide.ThepurityofthecruedCPwas34.43﹪thepurityoftherefinedCPwas79.68﹪.Thenucleinicacidpro