diagnostic mycology for lb t plaboratory ppf i · pdf file · 2014-01-26b....

Diagnostic ML b t PLaboratory P

Part OnePart One

Erik MErik MClinical Mi

Wheaton FranciWheaton FranciWauwatosa

The presenter states no conflict of inteto disclose relevant to the c

Mycology forP f i lProfessionalse--Yeaste Yeast

MunsonMunsonicrobiologyscan Laboratoryscan Laboratory, Wisconsin

erest and has no financial relationshipcontent of this presentation. 1

OUTL

I. Medical mycology

A. Comparisons wB. ClassificationB. Classification

II. Identification of clin

A. Cell morphologB Cultivation hintsB. Cultivation hintsC. Rapid and defin

III. Antifungal suscept

LINE

basics

with other life

nically-significant yeast

y hintsssnitive methods

tibility testing2

“D#*%it, Jim,I’m not a physician ”I m not a physician.

3

The BBasics

4

KINGDOM DIFFCharacteristic Fungi Plantae

Eukaryote Yes YesMembrane-bound

organelles Yes Yes

Non-mitochondrialib 80S 80Sribosomes 80S 80S

Chlorophyll None YesObligate

bNone None

anaerobes

Specialized tissue Sort of Higher plants

Sterols in membrane Yes Yes

Cell wallChitin, mannan, glucan, chitosan, CellulosCell wall g , ,

glycoprotein Cellulos

†Six-kingdom classification consists of either two Bacte

FERENTIATIONe Bacteria† Animalia

No Yes

No Yes

70S 80S70S 80S

Some NoSome No

No Yes

Mycoplasma Yes

e Peptidoglycan No iki die Peptidoglycan No en.wikipedia.org

eria kingdoms or two Protista kingdoms 5

SCOPE OAt least 100,000 nam

~1 million to 10 millio1000 to 1500 new sp1000 to 1500 new sp

Fewer than 500 namFewer than 500 namwith animal or human

Less than 50 are pathuman hostshuman hosts

Biol. Rev. 73:

OF FUNGImed fungal species

on unnamed species;pecies per yearpecies per year

ed species associateded species associatedn disease

hogenic in healthy

203-266; 1998 6

FUNCTIONALI

Pathogenesis (plants)Pathogenesis (plants)

Secondary metabolite

Symbiosis

HeterotrophyHeterotrophy

ITY OF FUNGI

))

es

7

FUNCTIONALI

Pathogenesis (humanPathogenesis (human-- Generally more chronic than

Secondary metabolite-- Generally involves predisposChemotherapy-induced neutropenia

Organ transplantation

SymbiosisOrgan transplantation

CorticosteroidsBroad-spectrum antimicrobials

P t l t iti

Heterotrophy

Parenteral nutritionDialysis

Invasive medical proceduresHeterotrophy-- Certain infections can be “sig

ITY OF FUNGI

ns)ns)acute

essitionHIV

DiabetesDiabetesAlcoholism

Intravenous drug abuseI t i l ti (b NICU)Intensive care population (burns, NICU)

MalignancyOther immune deficiency

gnal diseases”8

PAST TAX

Mode of reproductio

HolomTeleomorph

Sexual reproductionF i f t l i i t tFusion of two nuclei into zygote

Perfect FungiFilobasidiella neoformans

Type of sexual repro

XONOMY

n important

morphAnamorph

Asexual reproductionMit iMitosis

“Fungi Imperfecti”Cryptococcus neoformans

oduction important

yp

9

PAST TAXPhylum Zygomycota

Phylum B idi tBasidiomycota

Phylum Ascomycota

Phylum yDeuteromycota

XONOMYZygophores meet and fuse

(zygosporangium)( yg g )

Clamp connectionsfacilitate basidiumfacilitate basidium

Nuclear divisioni id (b )inside ascus (bag)

NO SEXUAL REPRODUCTIONOBSERVED 10

CURRENT TPhylum Zygomycota Subphy

RhizoRhizo

Subphy

Phylum Basidiomycota PhylumCrypto

Phylum Ascomycota Phylumy y y

Phylum Deuteromycota Not widelyy y t wid ly

TAXONOMYylum Mucoromycotinaopus Mucor Lichtheimia (Absidia)opus, Mucor, Lichtheimia (Absidia)

ylum EntomophthoromycotinaC nidi b lus Basidi b lusConidiobolus, Basidiobolus

m Basidiomycota h htococcus, Trichosporon anamorphs

m Ascomycotay80% of medically-important fungi

y utilized due to PCRy tili d d t P R

11

CLASSIFICATI

TaxonomyDisease

Deep seated (dissDeep-seated (dissOpportunistic myc

Subcutaneous mySubcutaneous my

Superficial mycoseSuperficial mycose

ION OF FUNGI

seminated) mycosesseminated) mycosescoses Aspergillosis

C didi iCandidiasisMucormycosis

ycoses Mycetomaycoses MycetomaChromoblastomycosis

es Dermatomycosis Tineaes Dermatomycosis TineaOnychomycosis Piedra

12

CLASSIFICATI

Taxonomy

Disease

Cell morphology (conBl tiBlastic blaEnlarge, then divide phi

Thallic art“Divide” then enlargeDivide , then enlarge

ION OF FUNGI

nidiogenesis)astoconidia annelloconidiaialoconidia poroconidia

hroconidia aleurioconidiachlamydoconidiachlamydoconidia

13

CLASSIFICATI

Taxonomy

Disease

Cell morphology (cop gy (

Colonial morphologyColonial morphology

YeastMo(u)ld

ION OF FUNGI

nidiogenesis)g )

yy

14

UNIFYING C

Microscopic observation

Direct detection

P(colony growth distr(colony, growth distr

CONCEPTS

n from primary specimens

n from primary specimens

Presumptive identificationribution cell morphology)ribution, cell morphology)

Rapid diagnosticsRapid diagnostics

Definitive identification15

The Preetenders

16

30-year-old MShortness of breath, pnight sweats poor oranight sweats, poor oraVitalsTmax 101°FTachypnea (22/min)Tachypnea (22/min)

StudiespO2 87% on room airCD4 count 17/mm3CD4 count 17/mm3

J. Clin. Microbiol

Male with HIVproductive cough,al intake weight lossal intake, weight loss

Tachycardia (95/min)Blood pressure 103/63Blood pressure 103/63

Albumin 2.9 g/dLBilateral diffuse infiltratesBilateral diffuse infiltrates

l. 49: 3113; 2011 17

Expectorated sputum

J. Clin. Microbiol

Bronchoalveolar lavage

l. 49: 3113; 2011 18

Pneumocystis jirovecProtozoan?? An

NoNo“Lif

F ?? CFungus?? Ch~AStaSta

Bacterium?? Trim

Different species infeNothing in vitro

cii (formerly carinii)tifungals ineffective

o ergosterolo ergosterolfe cycle”

itinAscospores internallyains with “fungus” stainsains with fungus stains

methoprim-sulfamethoxazole

ect different hosts

19

Pneumocystis jirovec

Serology, nucleic ac

Cyst wall, intracystic

Gomori methenamine silver

cii (formerly carinii)

id detection not useful

c body stains

Giemsa20

Pneumocystis jirovec

98.4% agreement be(histology) and fluoreantibody (microbioloantibody (microbiolo

J. Clin. Microbiol. 25

15% sensitivity in a cand induced sputumfrom AIDS populatiofrom AIDS populatio

Arch. Pathol. Lab. Med.

cii (formerly carinii)

etween toluidine blueescent monoclonalgy)gy)

5: 1837-1840; 1987

cohort of expectoratedm specimens collected

nn

. 112: 1229-1232; 1988 21

GeotrichYeast-like mold

Debilitated hosts; brorare disseminated disrare disseminated dis

Inhibited by cyclohexy y

hum spp.

onchial manifestations,seasesease

ximide; urease-negative; g

22

Prototheca wYeast-like achlorophy

Verrucous cutaneous Olecranon bursititsOlecranon bursitits

Sporangia withSporangia withseptations

Clin. Infect. Dis. 48

wickerhamiiyllous alga

infections

8: 1160-1161; 2009 23

“BLACK YAble to produce melasome (early) stage insome (early) stage in Most also produce tru

Many examples withinidentification based oMany examples withinBasidiomycota; lots o

Exophiala spp.Aureobasidium pullpHortaea werneckii

YEASTS”anized budding cells at

life cyclelife cycleue mycelium; therefore,

n Ascomycota andn asexual reproductionn Ascomycota and

of name changes

lulans24

“BLACK YSlow-growing; some Differential optimumE. jeanselmei my

Differential optimum j y

pha

E dermatitidis phaE. dermatitidis phapre

H. werneckii tine

A pullulans comA. pullulans compha

YEASTS”are very slow-growinggrowth temperatures

ycetoma

growth temperaturesyaeohyphomycosis

aeohyphomycosisaeohyphomycosisedilection for CNS; ocular

ea nigra

mmon contaminantmmon contaminantaeohyphomycosis 25

Exophiala jeanselmei

Aureobasidium pullulans

Exophiala dermatitidis

Hortaea werneckii 26

“BLACK Y

Organism Casein utilization

Tyrosine utilizationutilization utilization

E. jeanselmei - +E. dermatitidis - +H. werneckii + -

D. H. Larone, Medically Im

YEASTS”

Growth in 15% NaCl

KNO3assimilation

Maximum growth

temperatureNaCl assimilation temperature- + ≤ 37°C- - 42°C+ + Variable

mportant Fungi, fourth ed. 27

Ustilag“Corn smut fungus”

Inhaled and subsequisolated from sputumprarely implicated inhuman disease

White, pasty, moist,

human disease

p yyeast-like at first;becomes tan/brown,becomes tan/brown,mycelial within 20 day

go spp.

ently;;

ys28

The Conntenders

29

RhodotorProclivity to impact tecarcinoma and bactecarcinoma and bacteUrease-positive; rare

R mucilaginosa utilizpseudohyphaeR. mucilaginosa utiliz

rula spp.erminal stages oferial endocarditiserial endocarditise rudimentary

zes KNO ; R glutiniszes KNO3; R. glutinis -

30

SporobolomRare pathogenesis inpatientspatients

Best growth at 25°Cgkidney-shaped ballistocoloniescolonies

myces spp. immunocompromised

C; forcible discharge of; goconidia, forms satellite

31

MalassezNormal skin flora in m

Tinea versicolor; cradlCatheter-related sepsCatheter related seps

secondary pneum

Optimal recovery of thacquisition of blood via

Grows poorly at 25°C

acquisition of blood via

with thin layer of olive

zia furfurmore than 90% of adults

le cap; dandruffis (neonates TPN) withis (neonates, TPN) with

monia

he organism involvesa lipid infusion catheter

C; solid medium overlaid

a lipid infusion catheter

oil (not for veterinary)32

Malassez

Urease-

zia furfur

Tinea versicolor

Cells round at one endand bluntly cut at other

(no constriction);Collarette difficult toCollarette difficult to

discern by conventionalmicroscopy

-positive 33

SaccharomycTypically non-pathogeExposure to commerwith health foods andcolonization/infection


Bes

ces cerevisiaeeniccial strains associated

d baking may allow forg y

36: 557-562; 1998;

1-4 ascospores per ascusStain Gram-negative (vegetative

cells stain Gram-positive)Stain with Kinyoun stain (vegetativey ( gcells visualized with counterstain)

st demonstrated on specialized medium 34

TrichospoT. beigelii formerly coof genusof genusMajor taxonomic revis19 taxa recognized; ninfection caused by siinfection caused by siT. asahii T. asteroT cutaneum T inkinT. cutaneum T. inkin

Neutropenic patients;p p ;heart valve surgery; c

oron spp.nsidered main pathogen

sion in 1992nearly all systemicix speciesix speciesides T. mucoides

T ovoidesT. ovoides

AIDS; extensive burns;; ;catheterized patients 35

Trichospo

T. asachromogenic

T. asahiinutritive medium chromogenic

“Patriot B

nutritive medium

Urease-

oron spp.

ahiic medium

T. asahiicell morphologyc medium

Blue”

cell morphology

-positive 36

BlastoschizomObsolete taxa TrichosBl t hiBlastoschizomyces ps

Emerging cause of invEmerging cause of invin leukemic patients; mi i di hi hinvasive disease high

Difficult to delineate frDifficult to delineate fr

Urease-negativegNon-fermentative

myces capitatussporon capitatum and

d t i hseudotrichosporon

vasive fungal diseasevasive fungal diseasemortality rate fromi t i ti t in neutropenic patients

rom Trichosporonrom Trichosporon

Growth at 45°CGrowth on cycloheximide

37

“Current” DesignationClavispora lusitaniae teleomorphClavispora lusitaniae teleomorph

Yarrowia lipolytica teleomorphIssatchenkia orientalis teleomorph

Candida kefyrDebaryomyces hansenii teleomorph

Pichia guilliermondii teleomorphPichia guilliermondii teleomorphHansenula anomala (now obsolete)

Pichia anomala teleomorphWickerhamomyces anomalus

WickerhamWickerham

1-4 ascosp

Brim that tBrim that taround each a

“Former” DesignationCandida lusitaniae anamorphCandida lusitaniae anamorphCandida lipolytica anamorph

Candida krusei anamorphCandida pseudotropicalis

Candida famata anamorphCandida guilliermondii anamorphCandida guilliermondii anamorph

Candida pelliculosaHansenula anomala (now obsolete)

Pichia anomala

momyces speciesmomyces species

pores per ascus

turns downwardturns downwardascospore (helmet)

38

Candida Emerging opportunistPseudohyphae and trPseudohyphae and trelongate blastoconidiappearance; urease-


lipolyticatic pathogenrue hyphae bearingrue hyphae bearingia form stark branchingpositive


Candida lEmerging opportunishighly resistant to amhighly-resistant to amPseudohyphae slendshort chains of elong


lusitaniaest (malignancy);mphotericin Bmphotericin Bder, branched, curved;gate blastoconidia


CandidRare etiology of syste

Elongate blastoconid“logs in stream”g


da kefyremic disease, cystitis

dia line up in parallel;


Candid

e


da kefyr

Presence of metallicgreen sheen on Levinegreen sheen on Levine

eosin methylene blue (EMB)agar demonstrated 100%agar demonstrated 100%

positive predictivevalue for C. kefyr

0: 4281-4284; 2002 42

38-year-old Male w3-month history of GIfever chills poor appfever, chills, poor apploss over three monthPast contact with comCT imaging of abdomCT imaging of abdomfecal impaction; MRI intensity in periventricCSF protein 84 mg/d

intensity in periventricp g22 leukocytes (

ith GI & HeadacheI distress, headache;petite 20-pound weightpetite, 20-pound weighthsmmercial sex workermen consistent withmen consistent with

had increased signalcular and white matterdL (12-60) glucose 4 (40-70)

cular and white matter( ) g ( )

(0-5); 94% lymphocytes43

38-year-old Male wInitially reported as “

Tinkertoy® “appreciaamended reporta e ded epo t

Definitive identificatiCryptococcus neofo

Cryptococcal antigen

Patient also had CD

Cryptococcal antigen

load ~105 copies, th

ith GI & Headache“Abundant yeast”

ation”

on asrmans

n titer (CSF) 1024

4 count 29/mm3, HIV

n titer (CSF) 1024

,rush, Kaposi sarcoma

45

CryptococcusHIV signal disease; crAIDS-defining illness iCNS disease, skin, boAIDS-defining illness i

CSF cryptococcal antior exceeds that of cultor exceeds that of cult

S. Afr. Med. J. 71J Clin Microbiol 32

85.2% sensitivity of Giti f t


positive cases of cryptJ. Clin. Microbiol. 36

s neoformansryptococcosis is firstin 45% of AIDS patientsone, lung, other organsin 45% of AIDS patients

igen sensitivity equalstureture: 510-512; 1987 2: 1680-1684; 1994

ram stain in culture-t l i iti

2: 1680 1684; 1994

tococcal meningitis6: 1617-1620; 1998 46

CryptococC. neoformans/gattii c(e g C neoformans v(e.g., C. neoformans vSix species may be enC. neoformans C. aC. uniguttulatus C. luOnly C. neoformans/gphenol oxidase; “birds

g

phenol oxidase; birdstest utilizing caffeic acUrease-positive; nitrat

ccus spp.complex reclassificationvar neoformans)var. neoformans)ncountered clinically

albidus C. terreusuteolus C. laurentiigattii complex producesseed” agar also a rapidseed agar, also a rapidcid discte can help differentiate

47

NORTH AMERIC

PerSpecies Per

2008-2009C lbi 43 4C. albicans 43.4C. glabrata 23.5

C parapsilosis 17 1C. parapsilosis 17.1C. tropicalis 10.5

C krusei 1 6C. krusei 1.6† Estimate

J. Clin. Microbiol. 4J. Clin. Microbiol. 36

CA CANDIDEMIA

rcentage Distributionrcentage Distribution

1997 1952-1992†

55 3 54 055.3 54.017.0 8.012 1 7 012.1 7.07.2 25.02 3 4 02.3 4.0

49: 396-399; 2011 6: 1886-1889; 1998

49

CANDIDEM

Species Pe

0-19 yeyC. albicans 50C. glabrata 2.

C. parapsilosis 28C. tropicalis 12

C. krusei 0.

Diagn. Microbiol. Infect.

MIA BY AGE

ercentage Distribution

ear olds 80-99 year oldsy0.0 46.7.0 28.68.5 17.12.9 3.8.8 2.9

Dis. 68: 278-283; 2010 50

NOSOCOMIAL

Species Pe

ICC. albicans 50C. glabrata 17


C. krusei 2.

Int. J. Antimicrob. Ag

L CANDIDEMIA


CU Non-ICU0.4 47.47.5 18.15.1 18.90.5 9.6.1 2.1

ents 38: 65-69; 2011 51

EPIDEMIOLOGY

Species Pe

NosocomC. albicans 4C. glabrata 1


C. krusei 3

Antimicrob. Agents Chem

OF CANDIDEMIA


mial Onset Community Onsety7 518 188 151 113 1

mother. 55: 561-566; 2011 52

CandidaEmerging opportunistt fl lto fluconazole

Distinctive morphologDistinctive morphologchromogenic medium

Urease-positive (some

Elongate blastoconiditree-like appearance


tree like appearance

a kruseit; constitutively resistant

y ony on

e)

a,


Candida t

In patients with lympor leukemia, more vir

Distinctive morphologon chromogenic med

Blastoconidia singly

g

Blastoconidia singlyor in small groupsalong long pseudohyalong long pseudohy


tropicalis

horeticular malignancyrulent than C. albicans

gydium

yphaeyphaemportant Fungi, fourth ed. 54

Candida paInfections in particulcandidal endocarditicandidal endocarditiBlastoconidia singlycrooked/curved sho


arapsilosislary susceptible hosts;isis

y or in small clusters;rt pseudohyphae


Candida Potential induction of

b ti l tupon suboptimal trea

Endocarditis meningEndocarditis, mening

20% of Candida urina20% of Candida urina

When compared to CWhen compared to CC. glabrata CFU on eagar are larger

Diagn. Microbiol. Infec

agar are larger

glabrataf fluconazole resistancet tatment

gitis multifocal diseasegitis, multifocal disease

ary tract infectionsary tract infections

CFU on blood agar,CFU on blood agar,eosin methylene blue

t. Dis. 28: 65-67; 1997 56

Candida

Small blastoconidia;;may bud at 11:00 an

“Distinctive” morphoon chromogenic medon chromogenic med

R id t h l t tRapid trehalose testID within 3 hours whcell morphology; wat

glabrata

nd 1:00

logydiumdium

i i tiing gives presumptivehen correlated withtch out for blood agar

57

Candida Most common speciefrom all forms of canfrom all forms of canMake sure that germcontiguous (C. tropicconsidered presumpDistinctive morpholo

h i d

p p

on chromogenic medTerminal chlamydocoTerminal chlamydocoespecially at 25°C

albicanses isolated

ndidiasisndidiasism tube calis);ptiveogydi

p

diumonidia,onidia,

58

CHROMOGEN

Candida albicansCandida albicansCandida albicans

Candida tropicalisCandida tropicalisCandida tropicalis

J. Clin. Microbiol.J. Clin. Microbiol. 32

NIC MEDIUM

Candida kruseiCandida kruseiCandida krusei

34: 56-61; 1996 2: 1923-1929; 1994

59

Candida duMost frequently isolaHIV-positive patientsHIV positive patients oral candidiasis)Infrequently recovereInfrequently recoverevaginal specimens (imCandida albicans mim

germ tubechlamydCommercial germ tubfrequency of GT posi

germ tubechlamyd

frequency of GT-posiJ. Clin. Microbiol.

ubliniensisted from oropharynx of(pseudomembranous(pseudomembranous

ed from blood urineed from blood, urine,mmunocompromised)micrydoconidiabe reagent reducesitive C dubliniensis

doconidia

itive C. dubliniensis43: 2465-2466; 2005 60

TO ID OR N

C dubliniensis first deC. dubliniensis first deAIDS epidemic and inAs a result, mucocutacommonly seen; many ;resistant to fluconazo

NOT TO ID??

escribed in 1995 midstescribed in 1995, midstntroduction of HAARTaneous candidiasisny of first strainsyle (or inducible) 61

TO ID OR NOriginal C. dubliniensth t t i f i lthat yeast is fairly sus

Biochem. Soc. Trans. 3

Data corroborated byCDC 4.8%Denmark 3.1%Global surveillance 3.9%

J. Clin. Microbiol


J. Clin. Microbiol

NOT TO ID??sis discoverers note

tibl t fl lsceptible to fluconazole33: 1210-1214; 2005

y:% resistance in 42 isolates% resistance in 65 isolates% resistance in 310 isolates

. 49: 4415; 2011

8: 1366-1377; 2010 49: 325-334; 2011 . 49: 4415; 2011

62

DEFINITIVE IDECarbohydrate assimWickerham and Burtway to commercial kCandida sppCandida spp.Cryptococcus spp.Saccharomyces cerevisiaeRh d t l

PNA FISH f i

Rhodotorula spp.Trichosporon spp.

PNA FISH from posialbicans, C. glabratanot replace subcultu

ENTIFICATIONmilation is mainstay;

ton method has givengkits (some automated)

(former) Pichia spp(former) Pichia spp.Geotrichum spp.Malassezia furfurS b l

iti bl d lt (C

Sporobolomyces spp.Prototheca wickerhamii

itive blood cultures (C.a, C. tropicalis); doesure (polymicrobial)

63

MALDSpectra of 247/267 cliwith API biochemical pwith API biochemical pvery well (n = 220)


Prospective study of 6yielded 96.8% correctyielded 96.8% correct (P = 0.03 versus bioch84 0% correct ID to sp84.0% correct ID to sp


DI-TOFnical isolates correlatedprofiling; Big Five didprofiling; Big Five did

8: 2912-2917; 2009

61 yeast isolatesID to genusID to genus

hemical methods); peciespecies48: 900-907; 2010 64

Antifungal Susceeptibility Testing

65

CLSI DOCUMENT

M27 A3 ReferenceM27-A3 ReferenceDilution Susceptibili3rd ed. Approved S

M44-A2 Method forff SDiffusion Susceptib

2nd ed. Approved G

TS OF INTEREST

Method for Broth Method for Brothity Testing of Yeasts,

Standard

r Antifungal Diskfility Testing of Yeasts,

Guideline66

BROTH MICRCan test variety of yeaand C. neoformans, band C. neoformans, bInterpretive criteria on

Minimum InhAgent

Minimum Inh

Susceptible Susceptible(dose-depende

Fluconazole ≤ 8 16-32Fluconazole ≤ 8 16 32Voriconazole ≤ 1 2Itraconazole ≤ 0.125 0.25-0.50

5-fluorocytosine ≤ 4Anidulafungin ≤ 2Caspofungin ≤ 2Micafungin ≤ 2

CLSI M

RODILUTIONasts, including Candida

but not dimorphsbut not dimorphsnly for Candida spp.hibitory Concentration (g/mL)hibitory Concentration (g/mL)

nt) Intermediate Resistant Non-susceptible

≥ 64≥ 64≥ 4≥ 1

8-16 ≥ 32> 2> 2> 2

M27-A3 67

BROTH MICRRPMI 1640 broth (MO

24h Candida growth; Sabouraud dextrose

0.5 McFarland; dilutio

Sabouraud dextrose,

0.5 McFarland; dilutiorange of 0.5 x 103 to 2

Fluconazole, 5-fluorocytOther antifungalsOther antifungals

CLSI M

RODILUTIONOPS buffer, 0.2% dextrose)

48h C. neoformanspotato dextrose agar

on to final inoculum

potato dextrose agar

on to final inoculum2.5 x 103 CFU/mL

osine 0.12-64 g/mL0.03-16 g/mL0.03 16 g/mL

M27-A3 68

BROTH MICR35°C ambient air

24 hours for echin24-48 hours for am24-48 hours for flu48 hours for 5-fluo48 hours for other70-74 hours for C

Amphotericin B: obOther agents: obOther agents: ob

CLSI M

RODILUTION

nocandinsmphotericin Buconazoleorocytosiner azoles

C. neoformans testing

bserve 100% inhibitionbserve 50% inhibitionbserve 50% inhibitionM27-A3 69

DISK DIF

Has lagged behind br

Candida versus casflucfluc

ZoAgent

Zo

Susceptible Susce(dose-dep

Caspofungin ≥ 11

Fluconazole ≥ 19 15-Voriconazole ≥ 17 14Voriconazole ≥ 17 14-

CLSI M

FFUSION

roth dilution

spofungin posaconazoleconazole voriconazoleconazole voriconazole

one Diameter (mm)one Diameter (mm)

ptible pendent) Resistant Non-

susceptible≤ 10

18 ≤ 1416 ≤ 1316 ≤ 13

M44-A2 70

DISK DIFMueller-Hinton agar w(0 5 g methylene blu(0.5 g methylene blu24-hour Candida sppppDextrose agar0 5 McFarland standa0.5 McFarland standa1 x 106 to 5 x 106 CFCaspofungin 5 g Fluconazole 25 gFluconazole 25 g

CLSI M

FFUSIONwith 2% dextroseue/mL)ue/mL)

p. growth on Sabauroudp g

ard for inoc l m ofard for inoculum ofU/mL

Posaconazole 5 gVoriconazole 1 gVoriconazole 1 g

M44-A2 71

DISK DIF

35°C ambient air; 2

Observe for promineObserve for promine

Ignore pinpoint miIgnore large colon

CLSI M

FFUSION

20-24 hours

ent reduction in growthent reduction in growth

crocolonies at zone edgenies within inhibition zone

M44-A2 72

EteFluconazole, itraconastrips FDA approvedstrips FDA-approved Etest MIC data tendedmicrodilution for flucoCandida spp but oveCandida spp., but ove

Etest MIC data tendedmicrodilution for voricoCandida spp but ove


Candida spp., but ove

estazole, 5-fluorocytosinefor clinical usefor clinical used to be higher than brothgnazole testing of 1586

erall agreement 96 4%erall agreement 96.4%

d to be higher than brothgonazole testing of 1586erall agreement 98 1%1: 1440-1446; 2003

erall agreement 98.1%73

FLUCONAZOLE

Species P

19C. albicans 0.C. glabrata 8.

C. parapsilosis 0.C. tropicalis 0.

C. krusei 100


E RESISTANCE

Percentage Resistant

97 2008-2009.6 0.1.7 5.6.0 5.0.0 3.20.0 100.0

49: 396-399; 2011 6: 1886-1889; 1998

74

FLUCONAZOLE

Species P

NosocomC. albicans 0.C. glabrata 7.


C. krusei 100

Antimicrob. Agents Chem

E RESISTANCE


mial Onset Community Onsety.0 0.0.7 3.3.8 6.6.3 0.00.0 100.0

mother. 55: 561-566; 2011 75

FLUCONAZOLE

Species P

ICC. albicans 0.C. glabrata 5.


C. krusei 100

Int. J. Antimicrob. Ag

E RESISTANCE


CU Non-ICU.0 0.0.9 6.3.8 4.3.9 2.20.0 100.0

ents 38: 65-69; 2011 76

THE

Lots of name changeLots of name change

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diagnostic mycology for lb t plaboratory ppf i · pdf file · 2014-01-26b....

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