H1N2亞型豬流感病毒進化分析及抗HA蛋白單克隆抗體的制備.pdf

1、中國農(nóng)業(yè)科學(xué)院碩士學(xué)位論文H1N2亞型豬流感病毒進化分析及抗HA蛋白單克隆抗體的制備姓名：張飛申請學(xué)位級別：碩士專業(yè)：預(yù)防獸醫(yī)學(xué)指導(dǎo)教師：喬傳玲20080601AbstractSwineinfluenzavirus(SIV)isoneofimportantpathogensthatcauserespiratorydiseaseinpigsAtpresent，thesubtypesofSIVincludesHINI，H1N2，H1N7，H

2、2N3，H3N1，H3N2，H3N6，H4N6，H5NIandH9N2SeveralreassortantH1N2SIVstrainswereisolatedfromsomeprovincesinrecentyears，sincethefirstH1N2SIVstrainWasisolatedinChinain2004Whetherornotallepidemicswineinfluenzadiseaseinducedbythesevi

3、ruseswouldbreakOutinpigsshouldbefurtherstudiedInthepresentstudy，thespecificprimersforRT—PCRandsequencingweredesignedbasedonthepreferencesequencesofHlN2virusesAlltheeightgenefragmentsofSW／HeB／10／06andSW／T3／I／07aresequence

4、dandgeneticallyanalysisedwithbioinformaticsoftwaresResultsofgeneticanalysisshowedthatSW／HeB／10／06andSW／TJ／I／07arefarinrelativesHomologyofeightgenefragmentsofthetwostrainsis：PB2814％，PBl832％，PA83O％，HA901％，NP816％，NA917％，M84

5、1％，NS804％HAandNAgenefragmentsofSW／TJ／I／07showshighesthomologywithcorrespondinggenesofSW／ZJ／I／04(HIN2)，andotherinternalgenefragmentswithtWOHIN1SIVstrainsineuropeHA，NAandNSgenefragmentsofSW／HeB／10／06showshiighesthomologywi

6、thA／Turkey／MO／24093／99，andotherinternalgenefragmentswithHINIhumaninfluenzavirus，H3N2andH1N2swineinfluenzastrainsSW／HeB／l0／06iSprovedtobeanewreassortedvirusdifferentfromotherH1N2SIV，whichiSthefirstHIN2SIVincludinghumanavi

7、anandswineinfluenzagenesinmainlandWeproceedHItestamong4H1influenzavirusesandserumAccordingtotheresultsSW／HeN／II／05WasselectedtObepropagatedinembryonatedeggsAllantoicfluidswereharvestedandpuriedMixedwithFreundsadjuvantthe

8、viruseswereinjectedintoBALB／cfemalemiceTwomonoclonalantibody(McAb)hybridomacellstrainsweredevelopedbyfusingSP2／0cellswithspleencellsofmousethathadbeenimmunizedwithSIVSW／HeN／1I／05(H1N1)Clonedseveraltimes，positivehybridoma

9、cellsstrains4E682，3El1C4weredeterminedbyHIandELISAInHItest，ThetwoantibodiesshowspecificactivityinHI，noactivitywithH3，H5andH9swineinfluenzavirusandotherpathogensofswineInsummarygeneticrelationshipofthesetwoSIVstainswerean

10、alysedinthestudyTheresultsshowedthatthetwonewreassortedSIVstrainswerefarinrelativesThestrainsSW／TJ／I／07andSW／HeB／10／06providesevidenceforproducingnewreassortedinfluenzaviruseswithpigasavesselMeanwhile，twomonoclonalantibo