鹅细小病毒传代致弱毒株的培育与胶体金免疫层析检测方法的建立

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1、万方数据AbstractGooseparvovirusinfectioniscausedbygooseparvovirus(GPV)，whichisanacuteorsubacutesepsisin4to20days—oldgoslingscharacterizedbyhighinfectiousnessandexudativeinflammatoryboweldisease．Thisdiseasespreadsrapidlyandhasahighfatalityrate，themortalityrateofgosl

2、ingswithin5days．oldisabove95％．Thetransmissionandprevalenceofgooseparvovirusinfectionhascausedsevereeconomiclossesandrestrictedthedevelopmentofgooseindustry．ApathogenicagentisolatedfromgoslingswithpathogenicchangesandclinicalsymptomsofDerzsy’sdiseasebygooseembry

3、o，itwasidentifiedasgooseparvovirusandnamedasYGstrainafterPCRdetection，indirectimmunofluorescenceassay(IFA)detectionandanimalregressionexperiment．Toobtainattenuatedvaccinestrain，GPVYGstrainwaspropagatedalternatelyingooseembryofibroblasts(GEF)andduckembryofibrobl

4、asts(DEF)．GEFinfectedwithYGstrainbegantoinducecytopathogeniceffectafter12passages．IFAshowedthatthenumbersofinfectedcellswithgreenfluorescenceincreasedwiththetimeofinfection，andthehighestnumberofpositivecellswereobtainedat96hpost．infection．Virustitersincreasedwi

5、thserialpassagesgoingandvirustiterreachedat106·5TCIDsdmLafterpassage50．Comparedtoparentalvirus，themortalityandmorbidityofgoslingsdecreasedobviouslyafterinfectedwithcell．adaptedYGstrain．TherewerenoobservedcharacterizedpathogenicchangesorclinicalsymptomsofDerzsy’

6、sdiseaseingoslingsafterinfectedwithcell-adaptedYGstrain．FaecesofgoslingswhichinoculatedbyGPVF50weredetectedbyPCR，theresultsoffaecesdefecatedat3to21dayspost-inoculationwereGPVpositive，itshowsthattheattenuatedGPVcanpropagateinsusceptiblegoslings．Goslingsbegantopr

7、oduceneutralizationantibodyafterinoculatedbycell．adaptedYGstrain，andtheantibodytiterincreasedwiththetimeofinoculation．Theantibodytiterwas10。255／0．2mLat4weekspost-inoculation，thenitdecreasedslightly．ToidentifygeneticchangesoftheattenuatedGPV,thecompleteviralgeno

8、mesofGPVF50anditsparentalvirusFOweresequenced．Theresultsshowthattherewere89nucleotideand19aminoacidsmutationsbetweenthem．Therewere5aminoacidsmutationsinthenon·structuralprot