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1、JVetDiagnInvest7:301-304(1995)EvaluationofshellvialcellculturetechniqueforthedetectionofbovinecoronavirusRafiqueA.Tahir,KernA.Pomeroy,SagarM.GoyalAbstract.Theeffectofblindpassageandcentrifugationontheisolationofbovinecoronavirusinhumanrectaltumorcellsculturedinshellvialswasinvestigated.A
2、totalof68fecalsamplesknowntobepositiveforbovinecoronavirusbytransmissionelectronmicroscopic(TEM)examinationwereused.Thesampleswerecentrifugedontohumanrectaltumorcellmonolayersandincubatedinthepresenceoftrypsin.Thegrowthofbovinecoronavirusininfectedcellswasdemonstratedbyfluorescentantibod
3、ystaining,andtheextracellularviruswasdetectedandconfirmedbyhemagglutinationandhemagglutination-inhibitiontests,respectively.Ofthe68TEM-positivesamples,51(75%),58(85%),and61(90%)grewinshellvialcellculturesatfirst,second,andthirdpassages,respectively.Ofthe51culturespositiveonfirstpassage,1
4、9wereexaminedbyTEM;18ofthesewerepositiveforbovinecoronavirus.Theshellvialtechniquewasalsocomparedwithdirectdetectionofbovinecoronavirusbystainingcryostatsectionsofinfectedtissuesinadirectfluorescentantibodyassay.Theresultsofdirectfluorescentantibodyassaywereavailablefor54ofthe68samples,o
5、fwhich53(98%)and43(80%)werepositivebyshellvialtechniqueanddirectfluorescentantibodyassay,respectively.Foridenti-ficationofbovinecoronavirus,shellvialsusinghumanrectaltumorcellsinthepresenceoftrypsinismoresensitivethandirectfluorescentantibodyassaybutisrelativelylesssensitivethantransmiss
6、ionelectronmi-croscopy.23Bovinecoronavirus(BCV)is1oftheagentsthatvirusgrowspoorlyincellcultures.Atpresent,nocell13causesdiarrheaincalvesofupto4weeksofageandculturesystemisavailableinwhichBCVcangrowtohasfrequentlybeendetectedinthefecesofadultcattleasignificantlyhightiterduringprimaryvirus
7、isolation.1,20,21,2327withwinterdysentery.NeonataldiarrheacausedHumanrectaladenocarcinoma(HRT-18)cellsare5,14,29byBCVisresponsibleforheavyeconomiclossesinhighlysusceptibletoBCV,especiallywhentrypsin105,6bothdairyandbeefcattle.Inaddition,BCVcancauseisaddedtotheculturemediu
