志贺菌的多重pcr检测和分子分型研究-流行病与卫生统计学专业论文.docx

郑州大学2009届硕!一学位论文(摘要)志贺菌的多重PCR 检测和分子分型研究MultiPlex P C RD etection and MoleeularTyPin g aboutslligella，PPPostgraduate:X u Lanyin gSuPervisor:ProfessorXu BianliD ePartm ent ofEPidem iology，Zhengzhou U niversityZhengzhou，450001A bstraCtShigellae15thepathogenthatinducesbacilla尽办sente尽whichwaseonfirmedthe seco nd seriousinfeetion disease by the Chinese governm entin 1985.W ith theim Provem entoflivingstandardsandstrengtheningofm anagem ent，itsineideneerate15deelining，butstillm uehhigherthandeveloPedeountries.shlgellae’5Pathogenicityanddrugresistanee 15relative w ith itsvirulenee genes.Th erearem any researehesaboutviruleneegenesofShl’gellaeand theirdeteetion 15alsooneoftheroutinem onitoring，butm ostofthemeoneentrated on asinglePC R ，tim e一eonsum ing and laborious.A varietyofm olecularbiologytYPing m ethods，butnon ofthem 15standard.In view ofthis，thisexPerim ent15to develoP amethodofmultiPlexPCRtodeteetviruleneegenes(setjB，sen，切口从ial，stxlandvirA)inShl’gellasPPandeomPareseveralt即 ingm ethodsinorder tofindwhieh15m oresuitableformoleeulardeteetionofShl’gellae.M ethodsInorderto establishthe m ultiPlexPCR ，116 shigellaisolatesw erereeovered fromShigellam onitorsetsofHenanProvineeduringZOOI一2007(1strains.dys enteriae，114strainsS.刀ex月eri，11strains5.sonnei).Thesebacteriaarealso tyPed byrePetitiveextrageniePalindromiePCR(REP一PCR)，random amPlifiedPol卿orphieDNA (RAPD)，PlasmidProfile郑州人学2009届硕1学位论文(摘要)志贺菌的多重PC R 检测和分子分型研究andPulsed一fieldgeleleetroPhoresis(PFGE)teehniques and analyzedwithBioNum eriessoftware.R eSUltSEither6 virulenee genesw eredetected by multiPlex PC R atthe sam e tim e ordeteetedSeparatelyby generalPCR and itshow ed thatboth w ay sw ere sen sitive and sPeeifie.ThePositiveratesofsen，iPaH {stxl，l’al，virAandsetjB were88.70% ，99.13% ，0，88.70% ，92.17% and 85.22% resPeetively，and thenum beroftheirt即e w as 10.R EP 一PC R ’5Positiverate w as96.57%ando一IO b ands，w hieh ’s m oleeularw eightw as betw een l0ObP and2000bP，weream Plif