冠状病毒Coronavirusl论文-2016 [Springer Protocols Handbooks] Animal Coronaviruses __ Genotype-Specific Detection of Ferret Coronavirus by Convent.pdf

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1、Chapter16Genotype-SpecificDetectionofFerretCoronavirusbyConventionalandReal-TimeReverseTranscriptionPolymeraseChainReactionAnnabelG.Wise,MattiKiupel,andRogerK.MaesAbstractFerretcoronavirusisassociatedwithtwodiseasepresentationsinferrets,namely,epizooticcatarrhalenteri-tisandafelineinfectiousperiton

2、itis(FIP)-likesystemicdisease.Inthischapter,wedescribeconventionalandreal-timeone-stepreversetranscriptionpolymerasechainreactionassaysthatareroutinelyusedinourlaboratorytodetecteithergenotype1orgenotype2ferretcoronavirusinclinicalspecimens.Theseassaysweredesignedbasedupontheconservedspikegenesequ

3、encedifferencefoundbetweenthreestrainsofferretsystemiccoronavirusandthreestrainsofferretentericcoronavirus.Recentliteratureevidenceindi-catesthatpathotypeisnotassociatedwithaspeciÞcgenotype,andtherefore,itisimportanttotestforbothgenotypeseitherinentericorsystemicdisease.KeywordsFerretcoronavirus,C

4、onventionalRT-PCR,Real-timeRT-PCR,Moleculardiagnosticassay1IntroductionEpizooticcatarrhalenteritis(ECE),anentericdiseaseofdomesticferretswasÞrstdescribedintheUSAin1993[1].Clinically,thediseaseischaracterizedbyafoul-smellingbrightgreendiarrheawithhighmucuscontent,lethargy,anorexia,andvomiting.Morbi

5、dityapproaches100%,butmortalityisusuallybelow5%,withjuvenileferretsoftendevelopingonlymilddisease.ECEwasfoundtobeassociatedwithanalphacoronavirus,designatedferretentericcoronavirus(FRECV)[2].Thisdiseaseortheentericcoro-navirusagentassociatedwithithassincespreadintheUSAandothercountriesworldwide[3Ð

6、5].Closetoadecadeaftertherec-ognitionofECE,asystemicdisease,characterizedbyfelineinfec-tiousperitonitis(FIP)-likeclinicalsignsandlesions,begantoemergeinferretsacrosstheUSAandEurope[6Ð9].ClinicalÞnd-ingsincludeanorexia,weightloss,diarrheaandthepresenceofLeyiWang(ed.),AnimalCoronaviruses,SpringerPro

7、tocolsHandbooks,DOI10.1007/978-1-4939-3414-0_16,©SpringerScience+BusinessMediaNewYork2016171172AnnabelG.Wiseetal.largepalpableintra-abdominalmasses.AmarkedresemblancetoFIPisthegrossobservationofwidespreadwhitishn