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(xué)碩士學(xué)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(xué)位級別：碩士專業(yè)：預(yù)防獸醫(yī)學(xué)指導(dǎo)教師：胡永浩陳化蘭20050601甘肅農(nóng)業(yè)大學(xué)碩士學(xué)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