冠狀病毒Coronavirusl論文-2008 [Methods in Molecular Biology] SARS- and Other Coronaviruses Volume 454 __ Isolation and Propagation of Coronavirus.pdf

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1、10IsolationandPropagationofCoronavirusesinEmbryonatedEggsJamesS.GuyAbstractTheembryonatedeggisacomplexstructurecomprisinganembryoanditssupportingmembranes(chorioallantoic,amniotic,yolk).Thedevelopingembryoanditsmembranesprovidethediversityofcelltypesthatareneededforsucce

2、ssfulreplicationofawidevari-etyofdifferentviruses.WithinthefamilyCoronaviridae,theembryonatedegghasbeenusedasahostsystemprimarilyfortwogroup3coronaviruses,infectiousbronchitisvirus(IBV)andturkeycoronavirus(TCoV),butitalsohasbeenshowntobesuitableforpheas-antcoronavirus.IB

3、Vreplicateswellintheembryonatedchickenegg,regardlessoftheinoculationroute;however,theallantoicrouteisfavoredasthevirusreplicatesextensivelyinchorioallantoicmembraneandhightitersarefoundinallantoicfluid.TCoVreplicatesonlyinembryotissues,withinepitheliumoftheintestinesandb

4、ursaofFabricius;thusamnioticinoculationisrequiredforisolationandpropagationofthisvirus.Embryonatedeggsalsoprovideapotentialhostsystemforstudiesaimedatidentifyingother,novelcoronavirusspecies.Keywords:embryonatedegg;allantoic;amniotic;chicken;turkey;isolation;propagation;

5、diagnosis;detection.1.IntroductionEmbryonatedeggsareutilizedasalaboratoryhostsystemforprimaryiso-lationandpropagationofavarietyofdifferentviruses,includingthegroup3coronaviruses,infectiousbronchitisvirus(IBV),turkeycoronavirus(TCoV),andpheasantcoronavirus(1,2,3,4).Theyha

6、vebeenextensivelyutilizedforpropagationofthesevirusesforresearchpurposesand,inthecaseofIBV,forFrom:MethodsinMolecularBiology,vol.454:SARS-andOtherCoronaviruses,Editedby:D.Cavanagh,DOI:10.1007/978-1-59745-181-910,CHumanaPress,NewYork,NY109110Guycommercialproductionofvacc

7、ines.Inaddition,embryonatedeggsprovideapotentialhostsystemforstudiesaimedatidentifyingother,novelcoronavirusspecies.Theembryonatedeggcomprisesthedevelopingembryoandseveralsupport-ingmembranesthatenclosecavitiesorsacswithintheegg(5).Theshellmem-braneliesimmediatelybeneath

8、theshell;thisisatoughfibrinousmembranethatformstheairsacinthebluntendoftheegg.Incontrasttotheshellmembr