GP、GPM雙重PCR方法的建立及GPV VP3基因的克隆與序列分析.pdf

1、黑龍江八一農(nóng)墾大學(xué)碩士學(xué)位論文GP、GPM雙重PCR方法的建立及GPVVP3基因的克隆與序列分析姓名：姚笛申請(qǐng)學(xué)位級(jí)別：碩士專(zhuān)業(yè)：預(yù)防獸醫(yī)學(xué)指導(dǎo)教師：潘興廣崔玉東20060601AbstractGoslingP1ague(GP)isaTlacuteandsubacutesepticemicinfe吲ondiseasewhjchinducedbyG00separvovims(GPV)Since1997，theinfe謝0nwithGPMV

2、eruptedinourcountrywhichisch姍ct甜zcdbydjge鰣vediseasewithahighdegreeofmorbi擊tyandmon州吼whjchbmughtaserioushazardQn4quaculnlreGPa11dGPMhassimilarclinicalsymptoms，0nceinfectedmixedly，itisdifnculttoideIlti母a11ddia印osisfornocha

3、racteristicpath0109iedchaIlge1nordertoearlydiagnosisofclinicalcases，establisbingadisnn州ve，s朋sitive，andrapiddoublePCRdiagnos吐cmeIllodisofgreatsign訊canceTheartjclecam酣outcloningandsequencingAnalysisfor血eVP3oft11eGPVinHeil衄

4、面iaTl島wbichprovidesatheoretica】b商sfoT。diagllosisandprevemiona11dhasimport鋤tpradicalapplicationsofepidemiologicalsiglli6cance∞dvalue。Twopairsofspeci￡c州mersweredesjglledaccordingto廿leconserva廿vesequ鋤ceVP3geneofGPVandFgeneo

5、fGPMVinthegcneballl(theDNAofGPVandcDNAofGPMVin掣i行cialmixeds鋤pleswas鋤pJ語(yǔ)edbytlledoublePcRw油t11etwopairsprimersTheTcsultshowsthatthetwospecificbaIldsof574bp(GPV)aIld880bp(G孫Ⅳ)isconsistemwi廿1teStsdesiglled111edoublePcRwaSas

6、peci行csensitive鋤dspeedymethodfortlledetecti珈ofstaphylococcllsairellsenterotoxingenesThedeadyounggooseofHeiJon萄iangjn2005t嘣edwiththjsm舶od，the瞄u船shows廿1atmcsaIIlplesofsevencitjesareGPVin‰Ii伽andGPM|VisnotdefectedAccordillgt

7、oGPVB咖ing印omesequence，the州m盯ofVP3geneofGPVweredesi印edandsylltllesized，alld廿1esevencitiesGPvs鋤ples卸dVaccinejnhe訂0n鮒卸gwere跚pli6ed誦thPcR曲eresultobtainsⅡ鵬e矗砧ments，Ⅵ蚰chis574bp、477bp、699bprespectivelMAⅡdtlletllrec丘a鯽entswerecl

8、oneda11dtestedsequences，thellcomp州ngwi廿lissuedstmininco帥廿_yaIldabroadTheresuhsshdwesthat廿legeneofvP3is1605bp，whichencodes534枷inoacjdresiduessequenccda11alysisShowsthehomol090usofVP3geneinseveniSolatesis99O一998％andhonl010

9、90ustoisolateofNomleasta鰣culturalllnivers時(shí)is97498％Whichshowsstraillsisdi腩rentineachyear；homol090ustosYGstrainsis953—959％，講五chshowsitisdjff砷1tt0beMeent、voprovince；homol090ustoBs廿ajnsis964—971％W1lenconlparedwithvaccinestra

10、insthesamesol】rceis976983％GenesequellcingreSultSshowedVP3gelle3。曲dnucJeo硼eismoreconserv商vethen5’VP3IIdi嘶distllehi曲est—changingareasequellcing鋤al”isalsofoundfotlrsugarbasesitesofVP3protejnarehi曲lyc011servativeThus，tlleVP3

11、ofGPVsep眥tionmainsinHe柏on西iangisconseⅣatjve，t11isismolecueel鋤e11tsofonlyoneserotypeWhencomParedVP3genesoft11eseparationstrains、Ⅳith加)PVshowsthathom01090usis807_815％Keywords：Gooseparvovjms，GoosePar鋤y)(o橢s，doublePCRclone，s