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1、592BriefCommunicationsJVetDiagnInvest15:592±596(2003)Areversetranscriptase±polymerasechainreactionassayforthediagnosisofturkeycoronavirusinfectionBinuT.Velayudhan,Hyun-JinShin,VanessaC.Lopes,TomHooper,DavidA.Halvorson,KakambiV.Nagaraja1Abstract.Thisstudyreportsonthedevelopmentofareversetra
2、nscriptase±polymerasechainreaction(RT-PCR)forthespeci®cdetectionofturkeycoronavirus(TCoV).Oftheseveralsetsofprimerstested,1setofprimersderivedfromthePgeneand2setsderivedfromtheNgeneofTCoVcouldamplifytheTCoVgenomeintheinfectedsamples.TheRT-PCRwassensitiveandspeci®cforTCoVanddidnotamplifyoth
3、eravianRNAandDNAvirusestestedexcepttheinfectiousbronchitisvirus(IBV).ToovercometheproblemofIBVampli®cation,asetofseparateprimerswasdesignedfromthespikeproteingeneofIBV.TheRT-PCRunderthesameconditionsasabovecouldeffectivelydifferentiatebetweenTCoVandIBV.Thecloselyrelatedbovinecoronavirusand
4、transmissiblegastroenteritisvirusofpigsweredifferentiatedfromTCoVusingthesameRT-PCRwithslightmodi®cations.TheresultsofRT-PCRcorrelatedwellwiththeresultsoftheimmuno¯uorescenttestforthesamesamplestestedatthePurdueUniversityAnimalDiseaseLaboratory,WestLafayette,Indiana.Thenucleotidesequencean
5、dprojectedaminoacidsequencecomparisonofthePgeneofdifferentisolatesofTCoVfrom5differentstatesintheUnitedStatesrevealedacloseassociationamongthedifferentisolatesofTCoV.CoronavirusbelongstothefamilyCoronaviridae,atiousdiseaseaffectingturkeysofallages.Thediseaselargegroupofribonucleicacid(RNA)
6、virusesthatin-isknownbyvariousnames.Bluecombdisease,mudfectawiderangeofavianandmammalianspecies.fever,transmissibleenteritis,andinfectiousenteritisTheyarepleomorphicpositive-sensesingle-strandedaresynonymouslyusedtodescribecoronaviralenter-RNAvirusesmeasuring60±220nmindiameter.3,11itis.8,1
7、2ThisdiseaseisofhigheconomicimportanceThecharacteristicfeatureofthecoronaviralmorphol-becauseitcausespoorproductionperformance,im-ogyisthepresenceoflong(12±24nm),widelyspacedpairedgrowth,poorfeedconversion,andmortality.petal-orpear-shapedsurfaceprojections,whi
