Fluoromycobacteriophages for Rapid, Specific, and Sensitive Antibiotic Susceptibility Testing of Mycobacterium tuberculosis 英文参考文献.docVIP

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Fluoromycobacteriophages for Rapid, Specific, and Sensitive Antibiotic Susceptibility Testing of Mycobacterium tuberculosis 英文参考文献.doc

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Fluoromycobacteriophages for Rapid, Specific, and Sensitive Antibiotic Susceptibility Testing of Mycobacterium tuberculosis 英文参考文献

FluoromycobacteriophagesforRapid,Specific,and SensitiveAntibioticSusceptibilityTestingof Mycobacteriumtuberculosis MarianaPiuri1,WilliamR.JacobsJr.2,GrahamF.Hatfull1* 1Department of Biological Sciences, University of Pittsburgh, Pittsburgh, Pennsylvania, United States of America, 2Howard Hughes Medical Institute, Department of MicrobiologyandImmunology,AlbertEinsteinCollegeofMedicine,Bronx,NewYork,UnitedStatesofAmerica Abstract Rapid antibiotic susceptibility testing of Mycobacterium tuberculosis is of paramount importance as multiple- and extensively- drug resistant strains of M. tuberculosis emerge and spread. We describe here a virus-based assay in which fluoromycobacteriophagesareusedtodeliveraGFPorZsYellowfluorescentmarkergenetoM.tuberculosis,whichcanthen be monitored by fluorescent detection approaches including fluorescent microscopy and flow cytometry. Pre-clinical evaluationsshowthatadditionofeitherRifampicinorStreptomycinatthetimeofphageadditionobliteratesfluorescence in susceptible cells but not in isogenic resistant bacteria enabling drug sensitivity determination in less than 24hours. Detection requires no substrate addition, fewer than 100 cells can be identified, and resistant bacteria can be detected withinmixedpopulations.Fluorescencewithstandsfixationbyparaformaldehydeprovidingenhancedbiosafetyfortesting MDR-TBandXDR-TBinfections. Citation:PiuriM,JacobsWRJr,HatfullGF(2009)FluoromycobacteriophagesforRapid,Specific,andSensitiveAntibioticSusceptibilityTestingofMycobacterium tuberculosis.PLoSONE4(3):e4870.doi:10.1371/journal.pone.0004870 Editor:MadhukarPai,McGillUniversity,Canada ReceivedJanuary7,2009;AcceptedFebruary16,2009;PublishedMarch20,2009 Copyright:?2009Piurietal.Thisisanopen-accessarticledistributedunderthetermsoftheCreativeCommonsAttributionLicense,whichpermitsunrestricted use,distribution,andreproductioninanymedium,providedtheoriginalauthorandsourcearecredited. Funding:ThisworkwassupportedinpartbytheCenterforAIDSResearchattheAlbertEinstein