3株H9N2豬流感福建株的分離鑒定及分子衍化研究.pdf

1、甘肅農(nóng)業(yè)大學碩士學位論文3株H9N2豬流感福建株的分離鑒定及分子衍化研究姓名：趙有淑申請學位級別：碩士專業(yè)：預防獸醫(yī)學指導教師：胡永浩陳化蘭20050601甘肅農(nóng)業(yè)大學碩士學位論文英文摘要Isolation、IdentificationandMolecularEvolutionof3H9N2SubtypeSwineInfluenzaVirusesfromFujianProvinceSummarycandidate：ZhaoY0ushuA

2、dvisorProfHUYonghaoProLCHENHualanSwineinfluenza(SI)isapigrespiratorydiseasecausedbyswineinfluenzaviruse(SIV)Clinicalsignsincludehighfevers，coughing，dyspnea，muscleachingandcausingabortionsanddeathsamongthebreedingSOWSAtth

3、epresent，thediseasehavebecamewidespreadamongswineherdsintheworldPigsserveasanintermediatehostforgeneticreassortmentofhumanandavianvirusesbecausetheyCallbeinfectedbyeitheravianorhumanvirusesandaremorefrequentlyinvolvedini

4、nterspeciestransmissionofinfluenzaAvirusesthanotheranimalsInaddition，thesituationofavianinfluenzahasbecomemorecomplicatedinthemainlandofChinarecentlyTherefore，thestudingofthebiological，geneticpropertiesofthemainrecentepi

5、demicalstrainscirculatingamongChinapigswillnotonlyprovideusefulinformationforthediseaseprediction，promotehealthydevelopmentofthelivestock，butalsoprovideusefuldataforhumaninfluenzapandemicplan3swineInfluenzaviruseswereiso

6、latedfrom28nasalswabsobtainedfromFujianProvinceTheywereidentifiedasAinfluenzavirusesofH9N2subypcusinghemagglutinationinhibition(HAI)testsandneuraminidaseinhibition(NI)tests，andmolecularevolutionofthe3strainshasbeenanalyz

7、edinthisstudyViruseswereisolatedinembryonatedchickeneggsasdescribedinrefA33Eachviruswasbiologicallyclonedbythreeroundsoflimitingdilutioninembryonatedspecificpathogenfree(SPF)chicheneggsAfterthatallantoicfluidswereharvest

8、edStoretheisolatesat75℃Thenucleicacidsofthe3influenzaviruSeSconductedintheresearchwereextractedfromthevirusespropagatedinspecificpathogenfreechichenembryosAlloftheeightsegmentswereamplifiedbyRTPCR，andthepurifiedPCRproduc

9、tsweredonecyclesequencingwithspecificprimers，filrtherrglore，thesequencingproductswerepurifiedandrungelonCEQ8000DNAsequencingmachineThespecificprimersoftheeightgenesforPCRandcyclesequencingweredesignedusingtheOligo(60vers