Development of a Temperature-Switch PCR-Based SNP Typing Method for Mycobacterium ulcerans 英文参考文献.docVIP

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Development of a Temperature-Switch PCR-Based SNP Typing Method for Mycobacterium ulcerans 英文参考文献.doc

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Development of a Temperature-Switch PCR-Based SNP Typing Method for Mycobacterium ulcerans 英文参考文献

DevelopmentofaTemperature-SwitchPCR-BasedSNP TypingMethodforMycobacteriumulcerans KatharinaRo¨ltgen1,2,KobinaAssan-Ampah3,EmeliaDanso3,DorothyYeboah-Manu3,GerdPluschke1,2* 1SwissTropicalandPublicHealthInstitute,MolecularImmunology,Basel,Switzerland,2UniversityofBasel,Basel,Switzerland,3NoguchiMemorialInstituteforMedical Research,UniversityofGhana,Legon,Ghana Abstract Mycobacteriumulcerans(M.ulcerans),thecausativeagentofthedevastatingskindiseaseBuruliulcer(BU),ischaracterized by an extremely low level of genetic diversity. Recently, we have reported the first discrimination of closely related M. ulcerans variants in the BU endemic Densu River Valley of Ghana. In the study real-time PCR-based single nucleotide polymorphism(SNP)typingat89predefinedlocirevealedthepresenceoftenM.ulceranshaplotypescirculatingintheBU endemicregion.Herewedescribethedevelopmentoftemperature-switchPCR(TSP)assaysthatallowdistinguishingthese haplotypes by conventional agarose gel-based analysis of the PCR products. After validation of the accuracy of typing results,theTSPassaysweresuccessfullyestablishedinareferencelaboratoryinGhana.Developmentofthecost-effective andrapidTSP-basedgeneticfingerprintingmethodwillthusallowinvestigatingthespreadofM.ulceransclonesbyregular geneticmonitoringinBUendemiccountries. Citation:Ro¨ltgenK,Assan-AmpahK,DansoE,Yeboah-ManuD,PluschkeG(2012)DevelopmentofaTemperature-SwitchPCR-BasedSNPTypingMethodfor Mycobacteriumulcerans.PLoSNeglTropDis6(11):e1904.doi:10.1371/journal.pntd.0001904 Editor:PamelaL.C.Small,UniversityofTennessee,UnitedStatesofAmerica ReceivedJuly20,2012;AcceptedOctober1,2012;PublishedNovember15,2012 Copyright: ? 2012 Ro¨ltgen et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricteduse,distribution,andreproductioninanymedium,providedtheoriginalauthorandsourcearecredited. Funding:ThisworkwassupportedbytheStopBuruliInitiativefundedbytheUBS-OptimusFoundation.KatharinaRo¨lt