Mould Routine Identification in the Clinical Laboratory by Matrix-Assisted Laser Desorption Ionization Time-Of-Flight Mass Spectrometry 英文参考文献.docVIP

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Mould Routine Identification in the Clinical Laboratory by Matrix-Assisted Laser Desorption Ionization Time-Of-Flight Mass Spectrometry 英文参考文献.doc

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Mould Routine Identification in the Clinical Laboratory by Matrix-Assisted Laser Desorption Ionization Time-Of-Flight Mass Spectrometry 英文参考文献

MouldRoutineIdentificationintheClinicalLaboratory byMatrix-AssistedLaserDesorptionIonizationTime-Of- FlightMassSpectrometry CaroleCassagne1*,Ste′phaneRanque1,Anne-Ce′cileNormand1,PatrickFourquet2,SandrineThiebault1, ChantalPlanard3,MarijkeHendrickx3,RenaudPiarroux1 1Laboratoire de Parasitologie-Mycologie, CHU Timone, Universite′ de la Me′diterrane′e, Marseille, France, 2Service Prote′omique, Centre d’Immunologie de Marseille Luminy,Marseille,France,3IHEM:ScientificInstituteofPublicHealth,MycologyandAerobiologySection,Brussels,Belgium Abstract Background:MALDI-TOFMSrecentlyemergedasavaluableidentificationtoolforbacteriaandyeastsandrevolutionized thedailyclinicallaboratoryroutine.Butithasnotbeenestablishedforroutinemouldidentification.Thisstudyaimedto validateastandardizedprocedureforMALDI-TOFMS-basedmouldidentificationinclinicallaboratory. MaterialsandMethods:First,pre-extractionandextractionprocedureswereoptimized.Withthisstandardizedprocedure, a143mouldstrainsreferencespectralibrarywasbuilt.Then,themouldisolatesculturedfromsequentialclinicalsamples were prospectively subjected to this MALDI-TOF MS based-identification assay. MALDI-TOF MS-based identification was consideredcorrectifitwasconcordantwiththephenotypicidentification;otherwise,thegoldstandardwasDNAsequence comparison-basedidentification. Results: The optimized procedure comprised a culture on sabouraud-gentamicin-chloramphenicol agar followed by a chemicalextractionofthefungalcolonieswithformicacidandacetonitril.Theidentificationwasdoneusingareference databasebuiltwithreferencesfromatleastfourculturereplicates.Forfivemonths,197clinicalisolateswereanalyzed;20 wereexcludedbecausetheywerenotidentifiedatthespecieslevel.MALDI-TOFMS-basedapproachcorrectlyidentified 87%(154/177)oftheisolatesanalyzedinaroutineclinicallaboratoryactivity.Itfailedin12%(21/177),whosespecieswere notrepresentedinthereferencelibrary.MALDI-TOFMS-basedidentificationwascorrectin154outoftheremaining156 isolates.OneBeauveriabassianawasnoti