资源描述：
《冠状病毒Coronavirusl论文-1996 Factors Affecting Isolation and Propagation of Bovine Coronavirus in Human Rectal Tumor-18 Cell Line.pdf》由会员上传分享，免费在线阅读，更多相关内容在学术论文-天天文库。
1、JVetDiagnInvest8:96-99(1996)BRIEFCOMMUNICATIONSFactorsaffectingisolationandpropagationofbovinecoronavirusinhumanrectaltumor-18celllineSanjayKapil,KayL.Richardson,CraigRadi,CindyChard-BergstromBovinecoronavirus(BCV)isanimportantcauseofcalfultracentrifugedpelle
2、ts(approximately100,000xg),oras412enterocolitisandrespiratorydisease.Itisthesecondmajorpiecesofspiralcolon.Allsampleswerepositiveonlyforcauseofviraldiarrheaincalves,withrotavirusbeingtheBCVbyeitherdirectelectronmicroscopicexaminationor5first.AttheWisconsinAni
3、malHealthLaboratoryduringfluorescentantibodytest.1993-1994,BCVwasdetectedin93casesofcalfscoursoutFortrial1,fecalsuspensionsweretreatedwithdihydro-of1,058bovinefecalsamplesexaminedbydirectelectronstreptomycin(1mg/ml),penicillin(300units/ml),andfun-microscopy.E
4、lectronmicroscopyisusedcommonlyforthegizone(7.5µg/ml)for30minutesat37C.Sampleswerethen11bdiagnosisofentericviruses,includingBCV.TheadvantagesinoculatedontoHRT-18cellsgrownin12-wellplates.Forofelectronmicroscopyarethatdiagnosiscanbemaderapidlytrial2,approximat
5、ely1mloffecalsuspensionwasclarifiedandmultiplepathogens,acommonfeatureinenteritis,canbycentrifugationat1,000xgfor10minutes,diluted1:10bedetectedsimultaneously.Usingelectronmicroscopy,moreinminimumessentialmedium(MEM),andpassedthroughcthanadozennovelentericvir
6、useshavebeendescribedina0.45-µmfilter.Beforethesampleswerefiltered,themem-thelast2decades.However,electronmicroscopyhassomebraneswerewettedwithMEMtoavoidlossoftheviruslimitations;approximately1millionviralparticlesshouldthroughviralabsorptionandadsorption.bep
7、resenttodetectavirusbyelectronmicroscopy.Thus,itBeforeadsorptionofthevirus,HRT-18monolayerswere8lackssensitivityandcanleadtofalse-negativeresults.Inwashedoncewithcalcium-andmagnesium-freePBS(CMF-addition,someviruses,especiallycoronaviruses,canbecon-PBS)contai
8、ningtrypsin(5µg/ml).Afterthewashwasdis-fusedmorphologicallywithnonviralparticlessuchasintes-carded,theviruswasadsorbedfor1hourat37C.Samplestinalbrushborderepitheliumandwithothermorphologi
