简介：RapiddifferentialidentificationofMycobacteriumspeciesisessentialforeffectivediagnosisandmanagementofmycobacteriosis.Theaimofthisstudywastodevelopanovelmultiplexprobearraybasedonthe16S-23SrRNAgeneinternaltranscribedspacersequenceforthegenotypingofmycobacteriatothespecieslevel.Apairofprimersandasetofgenus-andspecies-specificprobesweredesignedfromtheconservedandpolymorphicregionsofthe16SrRNAgene,internaltranscribedspacer,and23SrRNAgenesequencesofmycobacteria.Weusedanovelmultiplexprobearrayforidentificationof266clinicalspecimensobtainedfrompatientswithmycobaterialinfection.Theresultsshowedthattheoverallspecificityandsensitivityofournovelprobearraywereboth100%forthegenus-specificprobeandMycobacteriumtuberculosiscomplex-specificprobe.Therewere79.3%(23/29)ofnontuberculousmycobacteriawhichcouldbeidentifiedtothespeciesleveldirectlyinthespecimensfromChina.SomeintraspeciesheterogeneityinM.avium,M.intracellulare,M.chelonaeandM.abscessuswasobserved.Withtheincreaseofsequencesofinternaltranscribedspacerandnumbersofwholemicrobialgenomes,andfurtheroptimizationofprobes,themultiplexprobearraywillbecomeapromisingtoolfortherapidandaccurateidentificationofmycobacteriainordinaryclinicallaboratories.

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简介：TheproteomicsofthedifferentialproteinexpressionsinhumangliomacelllineU251cellsinfectedwithhumancytomegalovirus(HCMV)wasinvestigatedattheproteinlevelbyusingthesurfaceenhancedlaserdesorption/ionization(SELDI)proteinchipsysteminordertodevelopamethodofstudyforthepathogenesisofHCMVinfection.Inthisstudy,theculturedU251cellswereinfectedwithHC-MVingoodconditionandthesupernatantsoflysatesandtheextracellularfluidsofthecultivatedinfect-edcellswerequantitativelydefinedfortheexpressedproteins.TheproteomicsofthedifferentialproteinexpressionincellsbeforeandafterinfectionwasanalyzedbyWCX2arraysontheproteinchipreader.Itwasdemonstratedthatthecytopathiceffectsofinfectedcellsappearedonthe5thdayafterinfection,however,thedifferentialproteinexpressionwasevidentat6hafterinfectionasrevealedbyRT-PCRandmassspectrometry.Theproteinpeakscapturedfromdifferentbatchesofsamples,fromthesamesampledetectedwithdifferentarraysorforthedifferentlimeswereallequivalent.Withthemolecularweightrangefrom2000Dato3000Da,chipcaptured82peaksfromtheintracellularfluidsand11proteinpeakfromthecellularfluidinwhichcomparedwiththecontrolgroup,theproteinpeakswithmolecularweightof13536.3Da,10046.1Daand17106.2Dawereclosetothoseofβ-amyloidpro-tein,caspase-1precursorandLPS-inducedTNF-αfactorrespectively,whichshowedbriefup-regulation4hafterinfection,andcontinuedtoraise48hlater.Theseresultsinferthattheseproteinsmaybere-latedtotheapoptosisinducedbyHCMVinfection,thussuggestingthattheapeptosisinducedbyHC-MVinfectionmayplayaroleinthepathogenesisofHCMVinfection.