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1、J.Vet.Med.B37,91-96(1990)01990PaulPareyScientificPublishers,BerlinandHamburgISSN0931-1793DepartmentofVeterinaryMicrobiology,IwateUniversity,Morioka,JapanPlaqueAssayandPropagationinRatCellLineLBCCellsofRatCoronavirusand5StrainsofSialodacryoadenitisVirusN.HIRANOA

2、ddressofauthor:Dr.NORIOHIRANO,DepartmentofVeterinaryMicrobiology,IwateUniversity,Morioka020,japanWith3figuresand3tables(ReceivedforpublicationJune28,1989)SummaryVariousfactorsinfluencingtheplaqueformationofratcoronavirus(RCV)andsialodacryo-adenitisvirus(SDAV)in

3、LBCcellmonolayerswerestudiedtodevelopthepracticalmethodforplaqueassay.Bythismethod,4JapaneseisolatesofSDAValsoproducedclearplaques.Inone-stepgrowthexperimentsoftheseviruses,newlyformedviruswasfirstrecognizedwithin7.5hpostinfec-tionandshowedsubsequentlyarapidexp

4、onentialincrease.Theviruswasreleasedrapidlyfromtheinfectedcells.Byindirectimmunofluorescencevirusspecificantigenwasdetectedasperinucleargranulesinthecytoplasmofthecellsat5-6hpostinfection,andallthecellsrevealedfluorescenceat12hpostinfection.Keywords:Ratcoronavi

5、rus,Sialodacryoadenitisvirus,LBCcells,Plaqueassay,ReplicationIntroductionRatcoronavirus(RCV)(6)andsialodacryoadenitisvirus(SDAV)(1)havebeenreportedtopropagatewithcytopathiceffect(CPE)onprimaryculturesofratkidneybutnotonanyestablishedcelllines(7).Inaddition,noad

6、equateplaqueassayforRCVandSDAVhasbeendescribed.Recently,theauthorsintroducedaratcellline,LBC,asausefulmediumforpropagationofRCV8190strain(2)andSDAV618strain(3),andreportedsuccessfulpropagationof4JapaneseisolatesofSDAVinLBCcellcultures(4).Ofseveraladvantagesofth

7、iscellsystemovertheprimarycultureofratkidney,themostsalientisthefactthatLBCcellsareverysensitivetothevirusandyieldhigh-titeredactivevirus.Inaddition,thecellscanbegrownandhandledeasily,andprovideasensitive,reproducibleassaymethodforinfectivity.Thepresentpaperdea

8、lswithinvestigationofvariousfactorsinfluencingplaqueformationinLBCcellswithRCVandSDAV,andastandardtechniqueofplaqueassayandthegrowthcharacteristicsofRCVand5strainsofSDAVincl