新生仔豬源大腸桿菌不同毒力因子環(huán)介導(dǎo)等溫?cái)U(kuò)增檢測方法的建立及應(yīng)用.pdf

1、厶類號＼㈣必必分類號Y32741Z9UDC——靜刪HOUu杰RS譬ITY碩士學(xué)位論文學(xué)號MZl4551密級(全日制專業(yè)學(xué)位)新生仔豬源大腸桿菌不同毒力因子環(huán)介導(dǎo)等溫?cái)U(kuò)增檢測方法的建立及應(yīng)用蔡樹東指導(dǎo)教師姓名：盛盤鎣塾撞：揚(yáng)劌盤鱟：婆蒸揚(yáng)劌：2至三QQ旦鲞蔓垂塾撞：婆蒸壅墼壁墊堅(jiān)些堂瞳：婆蒸查型：22三蘭QQ申請學(xué)位級別：亟學(xué)科專業(yè)名稱：獸醫(yī)碩士論文提交日期：2Q12生魚旦論文答辯日期：至Q12生魚爿學(xué)位授予單位：揚(yáng)劌太堂學(xué)位授予日期：2

2、Q!Z生魚旦答辯委員會主席：送繼遺II揚(yáng)州大學(xué)碩士學(xué)位論文AbstractNewbornpigdiarrheahasalwaysbeenanimportantdiseaseagainstthepigindustryalthoughthecausesofpigletsdiarrheaismorecomplex，pathogenicEscherichiacoliisanimportantcauseofpigletsdiarrheapatho

3、gensAtpresent，thedetectionmethodsofEscherichiacoliaremainlyisolatedandidentified，immuneserologicaltestsandtraditionalPCRmethods，butthesemethodsCannotmeettherequirementsoftherapiddetectionofgrassrootson—siteLoop—mediatedi

4、sothermalamplificationtechnologyisanewtypeofnucleicacidamplificationtechnologydevelopedbyNotomieta1Becauseofitssimpleopermion，fastandconvenient，highspecificity，highsensitivityandlowcost，ithasbeenwidelyusedinpathogenicmic

5、roorganismstestTherefore，tomeettheclinicalrapiddetection，thestudyestablishedtheLAMPtechnologyforthepigletpathogenicEscherichiacolidetectionandvirulenceanalysisLoop—mediatedisothermalamplificationtechnologyisdevelopedbyNo

6、tomietal，itisanewtypeofnucleicacidamplificationtechnologyBecauseofitssimpleoperation，fastandconvenient，highspecificityhighsensitivityandlowcost，inrecentyears，ithasbeenwidelyusedvirusdetection，parasitedetection，fungalandm

7、ycoplasmadetection，animalembryosexidentification，geneticallymodifiedfoodtestingandtumorgenedetectionTherefore，theestablishmentofLAMPtechnologytodetectEcoliforthepreventionandtreatmentofpigletsdiarrheaisofgreatsignificanc

8、eThisstudywasbasedontheconservedgenesofEscherichiacolienterotoxin(LTl，STl，ST2)gene，fimbriae(K88，K99，987P)gene，virulenceisland(HPI，ETT2，LEE)genepublishedbyGenBankSequence，asetofLAMPprimerweredesignedbyusingtheonlineprimer

9、designsoftware(http：／／primerexplorerjp／e／)TheconcentrationsofM92，theconcentrationofbetaine，theconcentrationofdNTPtheconcentrationofprimertheamountofBstDNApolymeraseandthereactiontemperatureandreactiontimewereoptimizedThe

10、LAMPtechnologyforEscherichiacolienterotoxingene，fimbriaegeneandirulenceislandgeneisestablishedInthisstudytakingenterotoxin(LTl，STl，ST2)genotype，fimbriae(K88，K99，987P)genotype，virulenceisland(HPI，ETT2，LEE)genotypeEcoliasp

11、ositive，eightcommonbacteriainclodingSalmonella，Staphylococcus，Riemerellaanat@estosis，Pasteurella，Streptococcus，Proteus，Clostridiumpe咿ingensandPseudomonasaeruginosaandultrapurewereusedasnegativecontrol，waterwereusedasblan