AgriculturalSciencesinChina2008,7(12):1516-1523©2008,CAAS.Allrightsreserved.PublishedbyElsevierLtd.December2008EstablishmentoftheMethodofImmunohistochemistryAssayfortheDetectionofScrapieinChineseShort-TailedHanSheepbyMonoclonalAntibodyZHANGYong-qiang1,WANGZhi-liang1,WUXiao-dong1,LIUYu-tian1,ZHANGHai-tao1,2andBAOEn-dong21NationalDiagnosticCenterforExoticAnimalDisease/ChinaAnimalHealthandEpidemiologyCenter,Qingdao266032,P.R.China2CollegeofVeterinaryMedicine,NanjingAgriculturalUniversity,Nanjing210095,P.R.ChinaAbstractThemethodofimmunohistochemistryassayforthedetectionofscrapieinChineseShort-tailedHansheepwasestablishedusingmonoclonalantibody.GenomicDNAwasisolatedfromChineseShort-tailedHansheepblood.Usingthepolymerasechainreactiontechnique,PrP27-30genesequencewasamplifiedfromChineseShort-tailedHansheepgenomicDNA.ByrecombinantDNAtechnology,therecombinantproteinofChineseShort-tailedHansheepPrP27-30wasobtained.Then,usingstandardmethodologyofmyelomacellfusion,apanelofmonoclonalantibodieswasgenerated.WithmAbs,scrapieinChineseShort-tailedHansheepwasdetectedbyimmunohistochemistryassay.TherecombinantproteinofChineseShort-tailedHansheepPrP27-30wasobtainedandapanelofsixhybridomacelllinessecretingspecificantibodiestoChineseShort-tailedHansheepPrP27-30relatedtoscrapiewasobtainedwithonefusionbetweenmyelomaSp2/0andspleencellsfrommiceimmunizedwiththepurifiedrecombinantprotein.FourhybridomacelllinescanbeusedinimmunohistochemistryassayforthedetectionofscrapieinChineseShort-tailedHansheep.Sothatthespecialmonoclonalantibodydevelopedinauthor’sinstitutecanbeusedtodetectPrPScofscrapieinChineseShort-tailedHansheepbyimmunohistochemistryinChina.Keywords:ChineseShort-tailedHansheep,PrP27-30,scrapie,monoclonalantibody,immunohistochemistryassayINTRODUCTIONTransmissiblespongiformencephalopathies(TSE)areneurodegenerativediseasesinhumanandanimals,in-cludingCreutzfeldt-Jakobdisease(CJD),fatalfamilialinsomnia(FFI)inhuman,scrapieinsheepandgoat,bovinespongiformencephalopathy(BSE)incow,andotherdisea...
