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1、JVetDiagnInvest10:93±97(1998)CorrelationofgenomicdetectionoffelinecoronaviruswithvariousdiagnosticassaysforfelineinfectiousperitonitisMelissaA.Kennedy,K.Brenneman,R.K.Millsaps,J.Black,L.N.D.PotgieterFelineinfectiousperitonitis(FIP)isafataldiseaseofdo-forRNAextraction.TheproceduresforRNAextract
2、ionmesticandnondomesticfelidscausedbyafelinecoronavi-weredoneasdescribed.b,3Approximately5mgofRNAwasrus(FCV).Itisasigni?cantproblemincatteries,multipleusedforRTwithMoloneymurineleukemiavirusreversecathouseholds,andshelters.14,18FIPcanmanifestasanef-transcriptasebbystandardmethods.13PCRwasperfo
3、rmedfusiveperitonitisand/orpleuritis,witharelativelyshortdis-withTaqpolymerasecalsobystandardmethods.13Thecon-easecourseendingindeath.Aprotractedcoursewithgran-centrationofprimerforRTandPCRwas0.5mM.Theam-ulomatouslesionsaffectingmultipleorgansmayoccur,pli?cationproductwaspredictedtobeapproxima
4、tely1ki-whichalsoinvariablyprogressestodeath.8,11,12lobase(kb)long.Theampli?cationproductfromthevirusTheFCVsarecloselyrelatedandincludetwobiotypes:propagatedinvitrowastheappropriatesize.thosethatarevirulentandcauseFIPandthosethatareavir-Toverifytheidentityoftheampli?cationproductandulent.7Thea
5、virulentgroup,knownasfelineentericcoron-con?rmthespeci?cityoftheseprimers,theDNAproductaviruses(FECV),maybeassociatedwithmildentericdis-fromampli?cationofstrainUCD1wasclonedintothep-easeorsubclinicalinfectionincats.11,12FCVsarealsoclas-CRIIvectorusingtheTAcloningmethoddaccordingtothesi?edintos
6、erotypes1and2basedonantigenicity.7,12Bothmanufacturer'sdirections.Thebasesequenceofclonedserotypescontainvirulentandavirulentbiotypes,andFCVscDNAwasdeterminedbythedideoxychainterminationwithinaserotypeareindistinguishableinthelaboratory.methodusing35S-ATPewithstandardmethods.13TheproductFCVsar
7、eubiquitous,especiallyinenvironmentssuchascat-had78%homologytothepublishedsequenceforthe7a/7bteries,wherelargenumbersofcatsarehousedtogetherandORFofFECVstrain79-1683(datanotshown).17Primers3,wherethemajorityofcatsareseropositive.12Asare
