基因芯片技術(shù)在食源性致病微生物檢測中的應(yīng)用研究.pdf

1、上海海洋大學(xué)碩士學(xué)位論文基因芯片技術(shù)在食源性致病微生物檢測中的應(yīng)用研究姓名：陳昱申請學(xué)位級別：碩士專業(yè)：食品科學(xué)指導(dǎo)教師：潘迎捷20090110上海海洋大學(xué)碩士學(xué)位論文ApplicationofDNAMicroarrayforDetectionofTheFoodbornePathogenicMicroorganismsABSTRACTArapid，sensitiveandspecificmethodofDNAmieroarraycomb

2、ined喇tllthePolymeraseChainReactionthatallowthesimultaneousdetectionandidentificationofSh／ge／／adysenteriaeStaphyloccocusaureusSalmonellaspp，Ecoli015ZVibriocholeraVibrioparahaemolyticusandListeriamonocytogenesThe16SrDNAgen

3、eandthekeypathogenicgeneswereusedastargetgenes，includingtheinvasionassociatedplasmidantigenHofSdysenteriae(ipaH)，thecoagulasegeneofStaphyloccocusaureus(coa)，Salmonellasppenterotoxingene(stn)andEscherichiacoli0157ShigaTox

4、ingene(slt)，thehemolysingeneofVibriocholera(^劬，thetoxRgenecodingTOXR—proteinofVibrioparahaemolyticusandtheclpEgenecodingclpE—proteinofListeriamonocytogenesThentheprimersandcapturedoligonucleotideprobesweredesignedandsynt

5、hesizedTheasymmetricPCRproductswerehybridized麗tllDNAmicroarray，whichcontainedspecificprobesofthesefoodbomepathogenicmicroorganismsOenomicDNAsfrom25bacterialstrainweredetected，theindexbacteriaswerepositiveanditonlyexistsf

6、alsepositiveamongSh／get／a@senteriae，SalmonellasppandEcoli01570ftheassaybased16SrDNAgeneThedetectionlimitofthismicroarrayassaywasaround8PggenomicDNAInaddition，thismethodWasappliedtodetectartificiallycontaminatedfoodsample

7、s，andthedetectionlimitwas50CFU／mLfornon—culturedsamplesWhileThedetectionlimitofthemultiplexPCRassayWasaround8PggenomicDNAand50CFU／mLfornonculturedsamplesTheseresultssuggestedthatdetectionofpathogenicmicroorganismsbyDNAmi

8、croarrayisaneffectiveprocedurewithhighspecificityandsensitivity，WhencomparedwiththeconventionalmicrobiologicalmethodsandothersTheDNAmicroarrayassaydevelopedinthisstudycouldprovideallinformativesupplementtoconventionalmic