Genetic Characterization of Type A Enterotoxigenic Clostridium perfringens Strains 英文参考文献.docVIP
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Genetic Characterization of Type A Enterotoxigenic Clostridium perfringens Strains 英文参考文献
GeneticCharacterizationofTypeAEnterotoxigenic
ClostridiumperfringensStrains
AgiDeguchi1.,KazuakiMiyamoto1.*,TomomiKuwahara2,YasuhiroMiki1,IkukoKaneko1,JihongLi3,
BruceA.McClane3,ShigeruAkimoto1
1DepartmentofMicrobiology,WakayamaMedicalUniversitySchoolofMedicine,Wakayama,Japan,2DepartmentofBacteriology,TheUniversityofTokushimaFacultyof
Medicine,Tokushima,Japan,3DepartmentofMicrobiologyandMolecularGenetics,UniversityofPittsburghSchoolofMedicine,Pittsburgh,Pennsylvania,UnitedStates
ofAmerica
Abstract
ClostridiumperfringenstypeA,isbothaubiquitousenvironmentalbacteriumandamajorcauseofhumangastrointestinal
disease,whichusuallyinvolvesstrainsproducingC.perfringensenterotoxin(CPE).Thegene(cpe)encodingthistoxincanbe
carriedonthechromosomeoralargeplasmid.Interestingly,strainscarryingcpeonthechromosomeandstrainscarrying
cpeonaplasmidoftenexhibitdifferentbiologicalcharacteristics,suchasresistancepropertiesagainstheat.Inthisstudy,
we investigated the genetic properties of C. perfringens by PCR-surveying 21 housekeeping genes and genes on
representative plasmids and then confirmed those results by Southern blot assay (SB) of five genes. Furthermore,
sequencing analysis of eight housekeeping genes and multilocus sequence typing (MLST) analysis were also performed.
Fifty-eight C. perfringens strains were examined, including isolates from: food poisoning cases, human gastrointestinal
diseasecases,foodsinJapanortheUSA,orfecesofhealthyhumans.InthePCRsurvey,eightofelevenhousekeepinggenes
amplifiedpositivereactionsinallstrainstested.However,byPCRsurveyandSBassay,onerepresentativevirulencegene,
pfoA,wasnotdetectedinanystrainscarryingcpeonthechromosome.Genesinvolvedinconjugativetransferofthecpe
plasmidwerealsoabsentfromalmostallchromosomalcpestrains.MLSTshowedthat,regardlessoftheirgeographicorigin,
dateofisolation,orisolationsource,chromosomalcpeisolates,i)assembleintoonedefinitiveclusterii)lackpfoAandiii)
lackaplasmidrelatedtothecpeplasmid.Similarly,independentoftheirorigin,strainscarr
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