冠状病毒Coronavirusl论文-2016 [Springer Protocols Handbooks] Animal Coronaviruses __ Antibody-Capture Enzyme-Linked Immunosorbent Assay for Detec.pdf

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1、Chapter5Antibody-CaptureEnzyme-LinkedImmunosorbentAssayforDetectionofAntibodytoTurkeyCoronavirusUsingInfectiousBronchitisVirusorRecombinantNucleocapsidProteinasCoatingAntigenChienChangLoa,MohamedAbdelwahab,Yi-NingChen,Ming-KunHsieh,ChingChingWu,andTsang

2、LongLinAbstractTurkeycoronavirus(TCoV)infectioncontinuestothreatenturkeyindustry.BecausespeciÞctreatmentandeffectivevaccinationprogramarenotavailable,rapidandcost-effectivedetectionofantibodiestoTCoVinfectionisanimportantcontrolmeasuretomonitorthediseas

3、estatusintheÞelds.Twoantibody-captureenzyme-linkedimmunosorbentassay(ELISA)proceduresfordetectionofantibodiestoTCoVareoutlinedinthischapter.OneELISAmethoduseschickeninfectiousbronchitiscoronavirus(IBV)asthecoatingantigenbasedonantigeniccross-reactivityb

4、etweenTCoVandIBV.TheothermethodreliesonarecombinantTCoVnucleocapsidprotein.BothmethodsareusefulforserologicaldiagnosisofTCoVinfectionintheturkeyßocks.KeywordsTurkeycoronavirus,Antibodydetection,ELISA,Nucleocapsid,Recombinantprotein1IntroductionTurkeycor

5、onavirus(TCoV)causesanacuteandhighlycontagiousentericdiseaseaffectingturkeysofallages[1,2].TCoVisanimportantpathogenandremainsasaseriousthreattotheturkeyindustry.ThereiscurrentlynospeciÞctreatmentoreffectivevac-cineforTCoVinfection.Inordertorapidlydiagn

6、oseandeffec-tivelycontrolturkeycoronaviralenteritis,serologicaldiagnosisofantibodiestoTCoVbecomesacriticaltooltodeterminetheinfec-tivityofturkeysintheturkeyßocks.Immunoßuorescentantibodyassay(IFA)fordetectionofanti-TCoVantibodieshasbeenextensivelyusedfo

7、rdiseaseoutbreaksintheÞelds.However,IFAproceduresaretimeconsumingandlaborintensiveforhandlinglargenumbersofsamples.Inaddition,IFArequireshighlytrainedpersonneltopreparefrozensectionsofLeyiWang(ed.),AnimalCoronaviruses,SpringerProtocolsHandbooks,DOI10.10

8、07/978-1-4939-3414-0_5,©SpringerScience+BusinessMediaNewYork20164344ChienChangLoaetal.turkeyorturkeyembryointestinesandperformßuorescentmicroscopy.ELISAmethodhashighspeciÞcityandsensitivityandcanbeappliedtolargenumbersofsamplessi