上海市臨檢中心 微生物學(xué)習(xí)班 11-MALDI TOF在臨床診斷方面2-劉朝陽

Porfortlio

of

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????剕???乓?????Microflex??????????????Z

?剕?????Z

????????A

True

Bench

Top

System????Full

Spectrum

ResolutionZ

News

from

ECCMID

2015Z

MBT???????????Z

MBT????????Z

ClinProt????Silent/Pleasant

OperationGreatly

Enhanced

SensitivityFast

Target

InsertionSelf

Cleaning

SourceSmartSpectra?AcquisitionFull

mass

spectrometerZ

MALDI??????13?剕??????М???????????????乎?????????Mass

spectrometry

(MS)is

an

analyticaltechnique

that

measuresthe

mass-to-chargeratio

of

charged

particles

in

gas

phase.??????????????????????????????????????????????X?????Bruker?BioSciencesNASDAQ:?BRKR?????お?X???????????????????????Whymassspectrometry?ZZZZZ????催???催?????????????MALDI親???????????????ˉ親????????????????????????–???????????????????剕????????????ぎ??Flex?MALDI?TOFSolarix

XR?FTMSGC/LC?QqTOF??????⑤????????????EIESIFAB……amaZon

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Laser

Desorption

/

Ionization????▔???(??)??MALDI

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10-7mbar催?ぎ10-7mbar??a-Cyano-4-hydroxycinnamic

acid(CCA,CHCA)Sinapinic

acid(SA)2,5-Dihydroxybenzoic

acid(DHB)2,5-Dihydroxyacetophenone(DHAP)+++m/z+++

++++++++2,4,6-Trihydroxyacetophenone(THAP)3-Hydroxypicolinic

acid(HPA)??IS1MALDI??⑤??親??trans-2-[3-(4-tert-Butylphenyl)-2-methyl-2-propenylidene]malononitrile

(DCTB)Trans-3-indoleacrylic

acid(IAA)Dithranol(DIT)親???

(???m/z)57????????親???????(????)?催?????:??:4-Hydroxy-?-cyanocinnamic

acid

(HCCA??-??-4-?????)2,5-Dihydroxyacetophenone(DHAP,

2,5-????Э?)Sinapinic

acid

(SA????)2,5-Dihydroxybenzoic

acid

(DHB)?宗:??:2,5-Dihydroxybenzoic

acid

(DHB?2,5-??????)3-Hydroxypicolinic

acid

(HPA?3-??-2-????)2,4,6-Trihydroxyacetophenone(THAP?2,4,6-????Э?)Dithranol,

HABA,

IAA,

DHB,

DCTB?????????????????▔???????М???????????????Thereflectoris

like

a

reverse

source,with

ionsgoingback

upa

potentialhillandturning

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moreenergeticionsgodeeperinto

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pathto

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SmartBeamTMMALDI-TOF(/TOF)??????▔???????MALDI????▔????????????????乥????????????????????ZZZ????????72)?72)????▔?????????乥??????じ?????????Smartbeam?▔???????????▔????▔????Smartbeam?▔???????▔?乥???1-2000Hz????*)

Ion

sourcebya

separate

turbopump

(not

shown).1

high

vacuum

ismaintainedHolleetal.,Journalof

Mass

Spectrometry,2006,41,705-71668MALDI-TOF??MBT

??????2015#

MALDI????????????????????????/??????#

催????????Bruker(20923712371+127+149?)bMX(?)??FDA??Ё?FDA??(RUO)1937552000

(???)#

?????????????????????г??????????????#

????∈⒊?????г????⒊??????????⒊???⒊?????Bruker(?)

Bruker(?)

bMX

(?)201320142015463156275989218522902371755755755#

催?????????(1ul??)#

?????MALDI????????????#

??∵??????#

????????????????Proteomics

LC-based???????MBT?????(2014)MALDI

????ぎ??????ProteomicsGel-based????????2014

(5627

MSPs)Biotyper????????1020

new

referencelibraryentries(main

spectra)from21

new

genera

and113

new

species50

new

bacterial

speciesin13

generaAlmost

doublingtheyeastlibrarywith63

newspecies

from

8

genera?????????????800

newentriesforbetter

coverage

ofdiversityinspecies

that

were

already

part

ofthelibrarybeforeImproveddifferentiationof

Haemophilushaemolyticus

vs.Haemophilus

influenza

(higherpathogenity)ClinProt?????????????TLC-MALDI?⒊??????Newstrainstoimprove

S.

pneumoniae

vs.

S.mitis/oralis

differentiation.??Additional

Nocardia

strainstocover

MALDIdiversityFor

RUO

MBT

and

IVD-MBTMALDI-TDS??????????GenoToolsSNP????/QC????911News

from

ECCMID

2015Norcardia

from

bMXNews

for

ECCMID

2015MBTsmartMBT?????(2015)The

High-end

Option

for

Laboratories

in

Leadership???2015

(5989

MSPs)?????New

patented

andproprietary

smartbeam

laser200Hzrepetiton

rate,i.e.>3

timesfasterlaserShoter

turnaround

time??362

new

MSPsZ271

improve

coverage

of

species

already

presentZ106

for

new

species10

new

genera

and

69

new

speciesLifetimelaser

with>

500M

laser

shotsFuture-proofforfurther

assayslike

MBT-STAR

BL

by

higherthroughputcapabilitiesMSP24322866626Genus19514347Species973???Higherlab

efficiency

by

reduced

down-timeduring

serviceFlexiblelabworkflows

byfaster

target

plate

exchangeGram??Gram?+Yeast1164192Thisis

NOT

a

successorofthemicroflex-LT-based

MBTbuta

high

endoption

for

laboratories

in

leadershipFil.?FungiSum652542??Not

IVDlabeled

inChina59894102371Pricing:+70k

Euro/80

k

USD

vs.classical

MBT1012Mycobacteria

Library

3.0Update

BenefitV2.0V3.0Mycobacteria

Library

3.0MALDI

SepsityperA

Quantum

Leap

in

Mycobacteria

IDBlood

CultureAnalysisMycobacteria

partners:NewAnkara,

TurkeyAlbany,

NY,

USABasel,Switzerland?542

MSPs!Z401fromclinical

isolates!Bern,SwitzerlandBorstel,GermanyChristchurch,NewZealandDessau,GermanyDortmund,GermanyBrest,FranceErlangen,GermanyGie?en,GermanyGroningen,

The

NetherlandsK?ln,Germany?21

species!Total??149

species!853

MSPs!ZZ408fromculturecollection

strains445from

well

characterized

clinicalisolatesLexington,KY,

USALuzern,SwitzerlandMadison,WI,USAMilwaukee,WI,USAMunich,GermanyNijmegen,

The

NetherlandsOslo,Norway?????90%???????

(165)VITEK

MS???????

50?

?Paris,FranceStra?burg,Franceand

DSMZ,

CCUG1315Mycobacteria

Library

3.0MALDI

Pilot149species

in

Mycobacteria

Library3.0,new

species

are

givenin

blue

fontM.abscessussubsp

abscessus

M.crocinum

M.koreenseM.porferaeM.M.M.M.M.M.M.M.M.M.M.M.M.M.M.M.M.M.M.M.M.M.M.M.M.M.M.M.M.aaaaaaaaaaaaaaaabbbbbbbbcccccMarch

2015CategoryM.M.M.M.M.M.M.M.M.cccccccccLifeScience

&

MedicineECCMID

2015

poster

1190,

Pranada

et

al.ticu.kns.oiSubcategoryLaboratory

Technology>700clinical

isolates

from

cooperation

partners

have

beenscreened

for

valuable

supplementation.445

MSPsincluded

from

well

characterized

clinical

isolates

provided

by

24

cooperation

partners

indifferent

countries.9Update

BenefitMBT

??乚?Typing/subtyping?Epidemology?Example

1:

Mycobacterium

immunogenum

(Wilson

etal.2001,

sp.

nov.)??Ubiquitous

environmental

organismPositive

BC

direct

IDIdentification

(Colony)Most

commonly

implicatedin

cutaneous

infectionin

both

healthyand

immunosuppressed

patients.

Also

known

to

cause

keratitis,hypersensitivity

pneumonitis,

bone

infections.Isolated

ID

technologyorCentral

platform??Resistant

to

many

anti-mycobacterial

agentsotherdirectID

&

selectivebroth-basedIDM.

immunogenum

MSPsZ

Mycobacteria

Library

2.0:

3Z

Mycobacteria

Library

3.0:

16Virus/SNP?Resistance?Virulence?1416MBT

??乚?MALDI

Sepsityper

Meta-AnalysisTyping/subtyping?Epidemology?Positive

BC

direct

IDIdentification

(Colony)Isolated

ID

technologyor??Sofar21

originalreportstestedtheSepsityperOverall,theSepsityperkitalloweda

reliableID

onthespecieslevelof

80

%

of

3320

positive

blood

culture

bottles.Central

platform?otherdirectID

&

selectivebroth-basedID??ID

rates:90

%

Gram-,

76%

Gram+

and

66

%

yeastIncombinationwithantibioticstewardship

programs,

thisrapidpathogenIDallowsa

muchfasteroptimizationofantibiotictherapyin

patientswithsepsis

comparedtoconventionalworkflows.Virus/SNP?ResistanceVirulence?MALDI

Sepsityper?

?Blood

CultureAnalysis????Reduced

timeforbacterialisolation.NoVacutainersorVacuettesnecessary.Only1

ml

bloodculturenecessary.Complete

workflowin

ONE

EppendorfCup.1719SepsisMS-Based

Resistance

Test??Sepsisis

a

clinicalsyndrome

thatcomplicatessevere

infection.????Typing/SubtypingMBT

STAR-BLMBT-RESISTItis

characterized

by

the

cardinal

signsof

inflammation

occurringin

tissuesthatare

remotefromtheinfection“Severe

sepsis”is

usedtodescribeadditional

disturbancesin

organfunctions“Septic

shock”is

the

most

severeclinicalsituationTheletality

increasesfrom

“sepsis”to“septicshock”????MBT-ASTRADespite

consensus

conferences,

theterminologyis

not

always

usedin

thesame

wayin

each

country?Hence,

there

are

not

always

the

samenumbers

reported

for

prevalence

ormortalityMALDI

SepsityperBenefiting

the

all

Hospital

StakeholdersSamples?run?directly?from?Positive?Blood?Cultures?

???????????Results?in?less?than?30?minutes?:?single?or?multiple?samples?run?in?parallelMaking?use?of?MBT?accuracy?with?full?library?access?in?same?time?to?run?Gram?StainMicrobiologist?&?ICU?Consultant?discuss?treatment?options?based?on?specific?patient?IDID?Results?to?ICU?Consultants?up?to?24?hours?quicker?than?traditional?methods75%?of?therapies?could?change?with?earlier?ID?:?reducing?overall?hospital?costs?????∈????90%???????????????3%JOURNAL

OF

CLINICAL

MICROBIOLOGY,

Nov.

2013,

p.

3711–37161820MSSA,

HA-MRSA??CA-MRSA?%The

performance

of

MALDI-TOF/MStodifferentiate

vanA-positiveEnterococcus

faecium(VPEF)fromvanA-negative

E.

faecium(VNEF)

was

evaluatedA

total

of

61

VPEF

isolates

and

71

VNEF

isolatesTo

discriminate

between

VPEF

and

VNEF,

allthree

ClinProTools

models

yielded

>90%recognitioncapability

(basic

sensitivity)

and

cross-validation

(reliability

of

the

models)The

geneticalgorithmmodel

exhibitedthehighest

performance

(99.18%

and

92.40%,respectively).The

high

detection

performance

of

MALDI-TOF/MS

for

VPEF

offers

the

potential

forroutine

laboratory

use.%%%%Satoshi

Nakano

etal.,2014(3),Pages

256–259Lu

etal.,Anal.

Chem.,

2012,

84,

5685?5692HA-MRSA??CA-MRSA?VNEF(61/63)Z

TAT

faster

than

that

of

thePCR

methodsZ

No

additionalcostZ

SimpleVPEF(55/59)Satoshi

Nakano

etal.,2014(3),Pages

256–259Lu

etal.,Anal.

Chem.,

2012,

84,

5685?56922123?

?MSSA??MSRA???????15/185???????⒊??????????121/127??????????????????Lu

etal.,Anal.

Chem.,

2012,

84,

5685?5692MSSA,

HA-MRSA??CA-MRSA?MBT

STAR-BL

WorkflowSelective

TestingofAntibiotic

Resistance

ofBeta-LactamaseID/optional

control

IDIncubation1-2h,

37?C1μl-loopfilledwithbacteriaCultureplateResuspensioninantibioticsolutionCentrifugationSupernatantSample:312

MRSAisolates109

MSSAisolatesTargetpreparationMBT

Compass

STAR-BL

moduleDataanalysisMALDI-TOF

MSSpectraacquisitionLu

etal.,Anal.

Chem.,

2012,

84,

5685?56922224MBT

STAR-BL

Principle?????Ampicillinnonhydrolyzed,

350.4,

372.4,

and

394.4

Da;hydrolyzed,

368.4,

390.4,

412.4,

and

324.4

Da;+18

Da-44

Dacefotaximenonhydrolyzed,

456.5,

478.5,

500,and

396

Da;hydrolyzed,

370

and

414

DaMass

shifts

by

metabolism

of

antibiotics

can

be

monitored

withthe

MALDIBiotyper!Peak

intensities

were

further

processed

using

the

formulalogRQ

=

log(sum[hydrolyzed

peakintensities])/(sum[nonhydrolyzed

peak

intensities]).Theresults

of

the

four

measurements

are

displayed

with

aboxplot

diagramConversion

ofAmpicillin

by

E.ColiAmpicillin

Hydrolysis

Assay

Result+Na+H2O+Na+Na+Na-CO230002500200015001000500E.

Coli

/Amp

cleavingstrain0600040002000E.

Coli

/DH5a03203303403503603703803904004104m20/z2527Cefotaxime

logRQ

valuesBlood

Culture??%Combined

usage

of

microbial

IDfrom

positive

blood

cultureswithMALDI

Sepsityper

and

subsequentMSBL

Assay

for

MBT-basedfunctional

resistance

testing.%Combined

result

2.5

hours

afterblood

culture

flagged

positive.%%Sensitivity:

100%Specificity:

91.5%

-100%(Depending

on

organisms)??Journal

of

Clinical

Microbiology

2014

Vol.

52(3)P924–930Cefotaxime

logRQ

valuessubcultures

on

Columbia

blood

agar???????????????

97%5???????

b

1

ml

??????剕?Sepsityper

Kit??

b??????b

??10

ul???⒊?(0.5

mg/ml

cefotaxime?????10mg/ml????)b

??b

37???90??

b

???????????

92%No

differences

inthe

logRQ

values

between

AmpC

hyperproducers

andESBL

positive

strains

were

observed

(BC

vs

subculture)5???????

b

????24??

(Columbia

blood

agar,MacConkey

agar,

and

Mueller-Hinton

agar)b

???????????∈?OD600

0.75

b

?1

ml???

b

??????

b

??10

ul???⒊?(0.5

mg/ml

cefotaxime?????10mg/ml????)

b??

b

37???90??

b

??????????55????

Escherichia

coliATCC

92112

(TEM

and

SHV

negative)??????Journal

of

Clinical

Microbiology

2014

Vol.

52(3)P924–9302628MALDI

Biotyper

STAR-BLMBT-RESISTHrabákJ,Walková

R,Studentová

V,

Chudácková

E,Bergerová

T.

(2011)

Carbapenemase

activitydetection

bymatrix-assistedExample

forpotentialautomateddatainterpretationlaserdesorption

ionization-time

offlightmass

spectrometry.

JClinMicrobiol.49:3222-7BurckhardtI,Zimmermann

S.

(2011)Using

matrix-assisted

laserdesorption

ionization-time

offlightmassspectrometry

to

detectcarbapenem

resistance

within1

to2.5

hours.JClinMicrobiol.49:3321-4.Calculation

of

the

normalizeddifference(S.

aureus)SparbierK,SchubertS,WellerU,

Boogen

C,KostrzewaM.(2012)MALDI-TOF

MSbased

functional

assay

forthe

rapid

detection0.8ofresistance

againstss-lactam

antibiotics.

JClinMicrobiol.50:927-37HrabákJ,Studentová

V,

WalkováR,Zemlicková

H,Jakubu

V,

Chudácková

E,

Gniadkowski

M,

Pfeifer

Y,

PerryJD,WilkinsonK,Bergerová

T.

(2012)Detection

ofNDM-1,VIM-1,KPC,

OXA-48,andOXA-162

carbapenemases

byMALDI-TOF

massspectrometry.

JClinMicrobiol.

[Epub

ahead

ofprint]Susceptible:NDiff

>0.5Kempf

M,

Bakour

S,Flaudrops

C,Berrazeg

M,

BrunelJM,DrissiM,

MesliE,

Touati

A,Rolain

JM.(2012).Rapid

detectionofcarbapenem

resistance

in

Acinetobacterbaumanniiusing

matrix-assisted

laserdesorption

ionization-timeofflightmass

spectrometry.

PLoS

One

7:e31676álvarez-Buylla

A,PicazoJJ,Culebras

E.(2013)Optimized

method

for

Acinetobacter

speciescarbapenemasedetection

andidentification

bymatrix-assisted

laserdesorption

ionization-time

offlightmass

spectrometry.

JClinMicrobiol.

51(5):1589-92Resistant:Wang

L,

Han

C,

Sui

W,

Wang

M,

LuX.(2013)MALDI-TOF

MSapplied

to

indirectcarbapenemase

detection:

a

validatedprocedureNDiff

<=

0.5to

clearlydistinguish

between

carbapenemase-positive

and

carbapenemase-negative

bacterialstrains.AnalBioanalChem.405(15):5259-66Lee

W,

ChungHS,Lee

Y,

Yong

D,Jeong

SH,LeeK,

Chong

Y.

(2013)Comparison

ofmatrix-assisted

laserdesorption

ionization-time-of-flightmassspectrometry

assaywithconventionalmethodsfor

detection

ofIMP-6,VIM-2,NDM-1,SIM-1,KPC-1,

OXA-23,andOXA-51carbapenemase-producingAcinetobacter

spp.,Pseudomonasaeruginosa,and

Klebsiella

pneumoniae.

DiagnMicrobiolInfectDis.[Epubahead

ofprint]JungJS,

Popp

C,SparbierK,

Lange

C,

Kostrzewa

M,

SchubertS.(2014)

Evaluation

ofmatrix-assisted

laserdesorption

ionization-timeofflightmassspectrometry

for

rapid

detectionof?-lactam

resistance

in

Enterobacteriaceae

derived

from

blood

cultures.J

ClinMicrobiol.52(3):924-30.Broad

antibiotic

resistancetesting

by

MALDI-TOF

MS?MBT-RESISTMBT-RESISTMBT-RESIST

assays

established

successfully??Staphylococcus

aureusOxacillin

(????),Cefoxitin(????)Ciprofloxacin

(????),Tobramycin(????),Meropenem(????)Pseudomonas

aeruginosa??Klebsiella

pneumoniaeE.

coliMeropenem(????)Cefotaxime(????)JOURNAL

OF

CLINICAL

MICROBIOLOGY,

Nov.

2013,

p.

3741–37482931Principle

of

the

MBT-RESISTMBT

-ASTRAMALDIBiotyperAntibiotic

Susceptibility

TestRapid

Assaynormal

LysControl1heavyLysControl2Journal

ofClinicalMicrobiology,

Dec.

2014,

p.

4155–4162Principle

of

the

MBT-RESISTMBT

–

ASTRAWorkflowAntibioticLysis

reagentwithinternalstandardBHIMcF0.5??????Incubation37eCSpeciesdependenttimeCelllysisTargetpreparationAcquisition

ofMSprofilespectraJournal

of

Clinical

Microbiology,

Dec.

2014,

p.

4155–41623032Principle

of

MBT

-

ASTRASpectra

viewStandard[M+H]2+Standard[M+H]+Standard[M+H]2+Standard[M+H]+ClinprotTM??????BHIMeropenem8

μg/mlBHIonlySusceptibleJournal

ofClinicalMicrobiology,

Dec.

2014,

p.

4155–4162ResistantData

evaluation

MBT-ASTRA???????????????CLINPROT:

Biomarker

Profiling2.Washing1.BindingArea

Under

Curve

??growthRelative

Growth3.ElutionPatientsSerumFractionationMALDI-TOFBioinformaticsResistantSusceptibleClinical

ResultClusterAnalysisPattern

DiscoveryNormalNormalDiseaseCluster

analysisNormalDiseaseBHIBHI+Ab*

**Journal

ofClinicalMicrobiology,

Dec.

2014,

p.

4155–41623335Titration

of

Meropenem

Conc.????????????????1.401.201.00ClinProt

BeadsClinProt

BeadsClinProt

Beads?∈?Klebsiella

pneumoniae????????????MB-HIC1MB-HIC3MB-HIC8MB-HIC180.800.600.400.200.00MB-WCXMB-WAXMB-IMAC-FeMB-IMAC-CuNRZ313-32NRZ0002-12K1-4NRZ1479-0.19PEG10489-0.032Inter-mediateSusceptibleResistant0.010.101.0010.00100.00ClinProt

BeadsClinProt

BeadsClinProt

Beadsμg/ml?Meropenem(DNA–

???)(??

??)(??-

??)Journal

ofClinicalMicrobiology,

Dec.

2014,

p.

4155–4162Analysis

of

Klebsiella

isolates??????(MB-WCX

??)8000600040002000For

TOF/TOF15\0_A15\1\1SLin,Smoothed,Baselinesubt.(97

K.

pneumoniae,

11

K.

oxytoca)800060004000200034\0_A16\1\1SLin,Smoothed,Baselinesubt.37\0_A17\1\1SLin,Smoothed,Baselinesubt.49\0_A18\1\1SLin,Smoothed,Baselinesubt.1\0_A19\1\1SLin,Smoothed,Baselinesubt.2\0_A20\1\1SLin,Smoothed,Baselinesubt.3\0_A21\1\1SLin,Smoothed,Baselinesubt.4\0_A22\1\1SLin,Smoothed,Baselinesubt.Conditions:1.601.401.201.000.800.600.400.200.00108

different

Klebsiella

isolates8μg/ml

Meropenem1hincubation

at

37eC72

susceptible8000600040002000800060004000200035

resistant8000600040002000Results:7500500025000.0010.010.11101005false

positives1false

negativeSpecificity:

93.5%Sensitivity:

97.3%MIC?[μg/ml]860004000200080006000420000020003000400050006000700080009000Journal

ofClinicalMicrobiology,

Dec.

2014,

p.

4155–4162m/z3436ClinProt??Complete

MALDI

Imaging

SolutionClinProt??Ultimateperformancein

MALDI

Imaging:?

20μm

spatialresolution(smartbeam-II)?

Upto

2kHzacquisitionspeed?

Laser-basedionsource

self-cleaningwithin15min3739ClinProt??????????????????????????????ぎ?????催?????????????????????

(???,

??,

????,

???….)?????????????????????ClinProt???????????????????Non-tumor???:Z?????Z???????Z????佄PTMsZ……tumor3840???????????Her2?ч??????ImmunohistochemistryInvasive

tumor

cellsCarinoma

insitu

(early

stage

tumor

cells)Neoplastic

cells

(pre-tumor

cells)Epithelial

cellsLymphocyte

infiltrationInflammationScoreScore2+Score3+0

and1+negativeequivocalpositiveConnective

TissueTumor

cells

are

not

homogenous:tumor

stem

cells(*)Different

stagesFluorescencein

situ

hybridizationDifferent

clonesnegativepositiveThe

surrounding

of

the

tumor

(stroma)influences

the

tumor:tumor

microenvirementMolecular

differentiationNo

TherapyTherapyTrastuzumab

(Herceptin?)????????????偠???????????????Discovery

setValidation

setPatients30132186??????????偠?????????ぎ?????IHCHer2

3+IHCHer2

2+IHCHer2

1+IHCHer2

-026???????????????????????????????131515141696FISH

Her2

positiveFISH

Her2negativeERpositiven.a.n.a.5???????????ぎ?????お???????????????ㄝ??????Н(MALDI????)ER

negativePRpositive13135PR

negative214143MALDI?????偠????????MALDIMass

SpectrometryTissueCryosectionMatrix

coatingm/z

4740m/z

8404AUC=0.84AUC=

0.930.0

0.2

0.4

0.6

0.8

1.00.0

0.2

0.4

0.6

0.8

1.0FalsepositiverateFalsepositiverateHER2-HER2+HER2-HER2+H&EstainingP

valueforeach<0.05MALDI????MALDI???????????J

Proteome

Res

2010;9(4):1854-63m/z

4969m/z

6225

m/z

4969m/z

8404Classification

of

HER2

receptor

statusin

breast

cancer

tissues

by

MALDI

imaging

massspectrometryRauser

S,

MarquardtC,BalluffB,

Deininger

SO,

Albers

C,

BelauE,

Hartmer

R,

Suckau

D,Specht

K,

Ebert

MP,

Schmitt

M,

AubeleM,H?flerH.

and

A.

Walch????Her2=

human

epidermal

growth

factor

receptor

type2Overexpression

in

20-30%

of

invasive

ductal

breast

cancersHer2

expressionis

correlated

with

poor

clinicalprognosisA

targeted

therapy

(trastuzumab,

monoclonal

antibody)

exists

for

the

treatment

ofHer2-positive

cancerm/z

4969m/z

6225

m/z

4969m/z

8404?Aim

of

this

study:

Proof

of

concept,

application

of

MALDI

imaging

toawell-known

clinical

system4244???????MALDI????IF

12Fractionsin

96-wellplate4.0

μL/well(