新生仔豬源大腸桿菌不同毒力因子環(huán)介導等溫擴增檢測方法的建立及應用.pdf

1、厶類號＼㈣必必分類號Y32741Z9UDC——靜刪HOUu杰RS譬ITY碩士學位論文學號MZl4551密級(全日制專業(yè)學位)新生仔豬源大腸桿菌不同毒力因子環(huán)介導等溫擴增檢測方法的建立及應用蔡樹東指導教師姓名：盛盤鎣塾撞：揚劌盤鱟：婆蒸揚劌：2至三QQ旦鲞蔓垂塾撞：婆蒸壅墼壁墊堅些堂瞳：婆蒸查型：22三蘭QQ申請學位級別：亟學科專業(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生魚爿學位授予單位：揚劌太堂學位授予日期：2

2、Q!Z生魚旦答辯委員會主席：送繼遺II揚州大學碩士學位論文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