猪附红细胞体分子诊断与致病机理的研究

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1、中国农业大学博士学位论文猪附红细胞体分子诊断与致病机理的研究姓名：贾杏林申请学位级别：博士专业：临床兽医学指导教师：齐长明;沈志强20050501AbstractThestudyfocusedonmoleculardiagnoseofEperythrozoonSuis，causeofPorcineEperythrozoonsis，theanimalmodelandnewtransmitwayandpathogenesisofthisdisease．1．ThestudyonmoleculardiagnoseofEpery

2、throzoonSuis：TheprimertodiagnoseEperythrozoonSuiswagdesignedaceofdingtotheforeignreportand781bpDNAfragmentwasamplifiedfromganomeofE．suisfrompig'sblood．TheDNAfragmentwaslinkedtopUCl8-TeasyvectorandclonedtoDHs-．TheplasmidwasidentifiedbyEcoRI肋fIandPastIdigestion．Th

3、ehomologybetweenthisfragmentandthatofE．suispublishedinGenebankis99．4％．Thereisn’thomologybetweenthisfragmentandothersequencepubfishedinGenebank．ThisDNAfragmenthash’tbeenamplifiedfromgnnomeofPasteurellosis，E。coil，StreptOCOCCUSSuis,Toxoplasm，TrichumsOvine，babesia’S

4、aimonellosis,Pseudorabiesvires,Porcinercpmduc'fiveandrespiratorySyndromeVirusandSPFpigblood．Thebloodsamples胁3farmsinwhichthepigadisplayedthesymptomofthisdiseaseand5healthfarmsweretestbyGimsadyeandmicroscopecheckandPCRdiagnose．Therearen’tdifferentbetweenmolecular

5、diagnoseandmicroscopechecktodetectEsuis．ThistestdemonstratedthatEperythrozoonSuispresentedinChinaandthepathogenyisthesanleasthatreportedinforeignbyPCP．diagnoseandthepathoganyisverypopularinShandongprovince．2．StudyonthemethodofdiagnosebyseparationE．suisfromerythr

6、ocyte：BeforeEPJL-1wasaddedintoblood，theerythrocytesofswinebloodweredeformedandlookedlikewave，starorsawbyE．soiswhenthebloodsmearswerestainedbyGimsadyeandobserved．TheE．suiswasstainedaspurplishredsmalldotaroundtheerythrocyte．TheerythrocytewasdeformedandtheE．suiswas

7、smallblackdotwhenitwasobservedunderOlympusmicroscope．AfterEPJL-1wasaddedintobloodandreacted0．5—1min，theerythrocytewasregularandthesurfaceoftheerythrocytewassmoothagain；moreovertheE．suisdidn’toccurredarounderythrocyte．10positiveswinebloodand1negativeswineblooddet

8、ectedbyPCRandmicroscopecheckwerepositiveandnegativealsobyseparationmethodrespectively．ThisseparationmethodcanseparateE．suis50reerythrocytewithin0．5·1rainandavoidblood