AUXACOLOR TM 2 5651320 tests

COLORIMETRIC SUGAR ASSIMILATION TEST FOR IDENTIFICATION OF THE MAIN YEASTS OF MEDICAL INTEREST

IVD

1- CLINICAL VALUEThe incidence of fungal infections, particularly those due to yeasts, hasmarkedly increased over the last twenty years and the profile of theseinfections, related to the growth in opportunistic infections has beenconsiderably modified.Due to the variability of the sites of these infections and the diversity of thespecies encountered, detection of yeasts in pathological specimens fromsuperficial or deep sites has become essential.Furthermore, in view of the recrudescence of these infections and themore widespread use of antifungal agents, resistant pathogenic strainshave emerged, which is why it is now essential to precisely identify theyeast species responsible for the infection.AUXACOLOR™ 2 meets this objective by identifying the species mostfrequently encountered in clinical practice. It is easy to use and tointerpret and can be easily integrated into all laboratories.

2- PRINCIPLE OF THE TESTThe AUXACOLOR TM 2 kit is an identification system based on theprinciple of sugar assimilation. The growth of yeasts is visualised by thecolour change of a pH indicator.The kit also includes 3 enzyme tests, including a test for detection of thephenoloxidase activity of Cryptococcus neoformans.

3- TYPES OF TESTS USED

The kit comprises :• a negative control to facilitate interpretation of the sugar assimilation

results (blue well).• 13 sugar assimilation results, corresponding to the following sugars:

- glucose (GLU.) : positive control- maltose (MAL.) - cellobiose (CEL.)- sucrose (SAC.) - trehalose (TRE.)- galactose (GAL.) - adonitol (ADO.)- lactose (LAC.) - melezitose (MEL.)- raffinose (RAF.) - xylose (XYL.)- inositol (INO.) - arabinose (ARA.)

2

Each sugar is dehydrated in the presence of a basic solution and a pHindicator, bromocresol purple. The growth of a yeast is indicated by thecolour change of the indicator from blue to yellow and by a cloudyappearance in the well.• An enzymatic test to detect N-acetyl-galactosaminidase activity

(hexosaminidase: HEX.). A positive reaction corresponds to a yellow colourof the well, while a negative test remains colourless.

• A phenoloxidase (POX.) test for detection of the phenoloxidase activityof Cryptococcus neoformans associated with a test to detect proline-arylamidase activity (PRO.) :- a brown colour of the well indicates positive phenoloxidase (POX)

activity.- a yellow colour indicates positive prolinearylamidase (PRO) activity.- absence of coloration or grey coloration indicates a negative

reaction for both tests.The coexistence of the POX and PRO tests in the same well can bejustified by the fact that these two tests are never both positive at the sametime. The only possible profiles are POX negative/PRO negative; POXpositive/PRO negative; POX negative/PRO positive with the colorimetricinterpretation described above.

MICROPLATE DIAGRAMAUXACOLORTM 2

BIO-RADLOT

REF

3

C.Neg GLU. MAL. SAC.

CEL. TRE. ADO. MEL.

GAL. LAC. RAF. INO.

XYL. ARA. HEX. POX./PRO.

4- COMPOSITION OF THE KITBox of 20 tests, code 56513, containing:• 20 x R1 individually packaged 16-well microplates• 20 adhesive films• 20 x R2 ready for use suspension media• 1 pad of results sheets• 1 package insert

5- WARNING AND PRECAUTIONS FOR USE• Do not use the kit after the expiry date.• Treat all samples as if they were potentially infectious and perform

cultures according to aseptic laboratory techniques.• Discard contaminated material in a container adapted to elimination of

laboratory waste.

6- STORAGE OF REAGENTSReagents must be stored between 2°C and 8°C in their individualpackaging. Under these conditions, reagents are stable until the expirydate indicated on the label.After opening the individual packaging of the microplate, it can be storedat room temperature for 10 hours with no alteration of performance.

7- PROCEDURE

Material suppliedSee Composition of the kit.

Material required, but not supplied• Opacity control kit, code 56499.• Loops for harvesting of colonies.• Pipettes to measure and deliver 100 µl.• Disinfectant bucket or autoclavable bag for elimination of contaminated

material.• Possibly a densitometer.

4

StepsAllow the reagents to adjust to room temperature before use.

1. Inoculation of the microplate- Prepare the inoculum from a

24 to 48 h culture on Sabouraudmedium (+/- antibiotics) orchromogenic medium(CandiSelect TM 4, code 63746).Under sterile conditions, inoculatethe suspension medium (R2) with asufficient quantity (1 to 5 identicalcolonies) of pure strain colonies(free of bacteria and other yeasts) toobtain an opacity equivalent to theopacity control supplied in the kitcode 56499 (opacity equal to 1.5 McFarland). Strict compliancewith the opacity of the inoculumdetermines the quality of the results.

- Homogenize the suspension in a vortex.

- Use a pipette to collect and distribute 100 µl of inoculum to each well of the microplate (R1).

- Cover the microplate (R1) with the adhesive film, ensuring perfectly uniformadhesion. Incubate for 48 hours (72 h if necessary) at 30°C (± 2°C).

2. Reading of the resultsThe definitive reading should be taken after 48 hours.Even though a first reading after 24 hours may give a correct score andallow certain yeasts to be identified, we recommend that definitiveinterpretation should be made after 48 hours.

5

. ... . . .. . .

C.Neg GLU. MAL. SAC. GAL. LAC. RAF. INO.

CEL. TRE. ADO. MEL. XYL. ARA. HEX. POX./PRO.

100 µ

l

suspension medium (R2)

If a Cryptococcus infection is suspected, the definitive reading should bemade after 72 hours given the slow growth of these microorganisms.Reading can be facilitated by looking through the underside of themicroplate or by removing the adhesive, respecting the usual conditionsof sterility if reincubation is necessary. In this case, carefully replace theadhesive, score the results with reference to the table on page 9 andrecord them on the results sheet designed for this purpose.

3. Interpretation of the results

a) Guide for interpretation of the colour reactions

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Well Test Colour/InterpretationNegative control C. Neg. Negative control Blue

Negative Positive

Suga

r as

sim

ilati

on t

ests

GLU Glucose (Positive control)

Blue (a)or

Green

Yellow (b)or

Colourless

MAL Maltose

SAC Sucrose

GAL Galactose

LAC Lactose

RAF Raffinose

INO Inositol

CEL Cellobiose

TRE Trehalose

ADO Adonitol

MEL Melezitose

XYL Xylose

ARA Arabinose

Enzy

me

test

s

HEXDetection of N-acetyl-

galactosaminidase activity(hexosaminidase)

Colourless Yellow

POX/PRO

Detection of thephenoloxydase activity ofCryptococcus neoformans

(POX)

Colourless or

Grey (c)

Brown

Detection of proline-arylamidase (PRO) activity

Yellow (b)

(a) Blue-grey and blue-green are considered as negative.(b) Pale yellow and yellow-green are considered as positive.(c) Some strains of Geotrichum capitatum, Geotrichum candidum,Trichosporon spp and Cryptococcus laurentii can cause a grey-browncoloration of the well. In this case the reaction is negative and the value"zero" should be given to this well when scoring

b) Scoring methodology

The 16 biochemical characteristics, distributed in 15 wells (the POX. andPRO. tests are combined in the same well), are used for identification.

• A 5-figure numerical profile is obtained by combining the values of thefollowing 15 tests by groups of 3:

A score of zero is attributed to each negative reaction and a scorerelated to its position in the triplet is attributed to each positive reaction:

The addition of the three scores gives a figure corresponding to a 5-figurenumerical profile:e.g. Glucose + Maltose + Sucrose ‡ 1 + 2 + 4 = the first figure is 7.• Proline-arylamidase activity (PRO: POX./PRO. well) is scored as + or -

according to the colour observed:- yellow well (pale yellow and yellow-green colorations are

considered as positive): PRO. positive test.

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1 for position 1 2 for position 2 4 for position 3

in the triplet

1st figure Glucose Maltose Sucrose

2nd figure Galactose Lactose Raffinose

3rd figure Inositol Cellobiose Trehalose

4th figure Adonitol Melezitose Xylose

5th figure Arabinose Hexosaminidase Phenoloxidase

- colourless or grey well (grey brown coloration is considered asnegative): PRO. negative test.

• Two additional figures are calculated using the method describedabove to complete the score. They represent the following characters:

The final identification is based on a combination of the biochemical testsand complementary criteria (morphological and metabolic) establishedunder the usual conditions.Examples:

Calculation of the corresponding numerical profiles and identification:

After the numerical profile has been determined, this is sought in the databasegiven on page 15.If the numerical profile obtained is not indicated on the list (rare profile) referto the interpretation table (page 9).

8

Pigmentation (PI.) Arthrospores (AR.) Capsule (CA.)

Mycelium/pseudo-mycelium (MY. PS-MY.) Chlamydospores (CHL.) Growth at 37°C

STRAIN GLU. MAL. SAC. GAL. LAC. RAF. INO. CEL. TRE. ADO. MEL. XYL. ARA. HEX. POX./PRO. PI. AR. CA. MY./PS-MY. CHL. 37°C

Strain N°1 + + + + - - - - + - - + - + - + - - - + + +Strain N°2 + - - - - - - - + - - - - - - - - - - - - +Strain N°3 + + + + - + + - - + + - - - + - - - + - - +

StrainNumerical profile

IdentificationBiochemicaltests

PRO testAdditionalcharacters

Strain N°1 71442 + 07 C. albicans

Strain N°2 10400 - 04 C. glabrata

Strain N°3 75134 - 44Cryptococcusneoformans

9

INTE

RPRE

TATI

ON

TA

BLE

GLU

.M

AL.

SAC.

GA

L.LA

C.RA

F.IN

O.

CEL.

TRE.

AD

O.M

EL.

XYL.

ARA

.H

EX.

POX.

/PRO

.PI

.A

R.CA

.M

Y.CH

L.37

°CSP

ECIE

SPS

-MY.

C. A

LBIC

AN

S 1

++

++

--

--

+ (-)

VV

+ (-)

V+

(-)-

+-

--

++

(-)+

C. A

LBIC

AN

S 2

(1)

++

-+

--

--

VV

-V

V+

--

--

-+

+ (-)

+C

. CIF

ERRI

I+

++

+-

+ (-)

+V

++

-+

+V

--

--

-+

-+

C. D

UBL

INIE

NSI

S+

++

+-

--

-V

+ (-)

- (+)

--

+ (-)

-+

--

-+

++

C. F

AM

ATA

++

++

V+

-+

++

+ (-)

VV

--

V-

--

--

VC

. GLA

BRAT

A+

--

-- (

+)-

--

+-

--

--

--

--

--

-+

C. G

UILL

IERM

ON

DII

++

++

-+

(-)-

++

+ (-)

+ (-)

+ (-)

+-

-+

--

-+

-+

C. I

NC

ON

SPIC

UA

+-

--

--

--

- (+)

--

--

--

V-

--

--

+C

. KEF

YR+

-+

++(

-)+

-V

- (+)

- (+)

-V

V-

--

--

-+

(-)-

+C

. KRU

SEI

+-

--

--

--

--

--

--

-- (

+)-

--

+-

+C

. LIP

OLY

TIC

A+

--

- (+)

--

-- (

+)-

- (+)

--

--

-+

(-)-

--

+-

VC

. LU

SITA

NIA

E+

++

+ (-)

- (+)

--

++

V+

+ (-)

V-

-V

--

-+

(-)-

+C

. NO

RVEG

ENSI

S+

--

--

--

+ (-)

--

-- (

+)-

--

+-

--

+-

+C

. PA

RAPS

ILOSI

S+

++

+-

--

-+

(-)V

+ (-)

+ (-)

+-

-V

--

-+

-+

C. R

UG

OSA

+-

-+

(-)-

--

--

V-

V- (

+)-

-V

--

-+

-+

C. S

AKE

++

+V

--

-V

+V

+V

-+

(-)-

V-

--

+-

-C

. TRO

PIC

ALIS

++

+ (-)

+-

--

V+

+ (-)

++

--

-- (

+)-

--

+-

+C

. ZEY

LAN

OID

ES+

--

- (+)

--

-- (

+)+

(-)V

--

-- (

+)-

+ (-)

--

-+

-- (

+)C

. ALB

IDU

S+

++

VV

VV

+V

V+

+V

--

V-

-+

--

VC

. LA

URE

NTI

I+

++

++

++

++

++

++

--

--

-+

--

-C

. NEO

FORM

AN

S+

++

+-

V+

VV

V+

(-)V

V-

+ (-)

--

-+

--

+C

. UN

IGUT

TULA

TUS

++

+V

-V

+-

VV

++

(-)V

--

V-

-+

--

-G

. CA

ND

IDU

M+

--

+ (-)

--

--

-V

-+

--

- (+)

- (+)

-+

-+

-- (

+)G

. CA

PITA

TUM

+-

-V

--

--

--

--

--

--

-+

-+

-+

K. A

PIC

ULA

TA+

--

--

--

+-

--

--

--

--

--

--

-R.

GLU

TIN

IS+

++

V-

V-

V+

V+

VV

V-

++

--

--

VR.

MUC

ILAG

INO

SA (R

UBRA

)+

V+

V-

+-

VV

VV

VV

V-

++

--

--

VS.

CER

EVIS

IAE

++

(-)+

V-

+ (-)

--

V-

V-

--

--

--

-- (

+)-

VT.

ASAH

II+

+V

++

-V

+V

VV

VV

+ (-)

--

-+

-+

-+

T. IN

KIN

++

+V

+-

+ (-)

++

(-)-

++

V+

(-)-

--

+-

+-

+T.

MUC

OID

ES+

++

++

++

++

+ (-)

++

+V

--

-+

-+

-+

T. SP

P+

++

(-)+

(-)+

VV

++

(-)V

V+

(-)+

(-)V

-- (

+)-

+-

+-

VP.

WIC

KERH

AMII

(2)

+-

-+

(-)V

--

-+

--

--

--

--

--

--

+

CAPTION(1) C. albicans 2: corresponds to strains formerly classified under thespecies C. stellatoidea, rarely encountered.(2) P. wickerhamii: this is a pathogenic unicellular alga, which grows easilyon yeast isolation media and presents a similar macroscopic appearanceto that of yeasts. Microscopic examination reveals round cells, dividing byinternal septation.

+ : Test positive for at least 99% of strains

- : Test negative for at least 99% of strains

V : Variable test

+(-) : Test positive for at least 95% of strains or delayed expression test(48 hr or 72 hr)

-(+) : Test negative for at least 95% of strains

PI. : Pigmentation

AR. : Arthrospores

CA. : Capsule

MY. : Mycelium (P.C.B. medium)

PS-MY. : Pseudo-mycelium (P.C.B. medium)

CHL . : Chlamydospores

37°C : Growth at 37°C

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c) Notes

1- Identification of yeasts is based on both their biochemical charactersdetermined on the AUXACOLOR ™ 2 microplate and on theirmorphological characters, preferably determined on PCB medium orpossibly on RAT medium. All of these data must be obtained forcomplete identification. The origin of the samples and the clinical context must also be takeninto account in interpreting the results.

2- The species Rhodotorula glutinis and Rhodotorula mucilaginosa areindicated in the interpretation table, but their numerical profiles are notspecified, as many of their characters are variable. These 2 species arecharacterised by a salmon pink pigmentation of the colonies, whichoften appears after 48 h of culture.

3- Yeasts of the Cryptococcus genus form a capsule which may be verydiscrete.

4- The morphology of the yeast Kloeckera apiculata is characteristic,allowing it to be distinguished from species with the same numericalprofiles: very small, elongated yeast, with bipolar buds (resembling alemon).

5- It may be useful to perform supplementary tests to confirm theidentification of some species of yeasts or to distinguish species withthe same numerical profiles:- the potassium nitrate assimilation test can distinguish yeasts such as

Cryptococcus albidus (positive test) and Cryptococcus laurentii(negative test) or Rhodotorula mucilaginosa (negative test) andRhodotorula glutinis (positive test).

- the test for urease on urea-indole medium confirms identification of theCryptococcus genus. This test is positive in 4 hours for Cryptococcusneoformans, and in 24 hours for the other species.

- the reduction of tetrazolium distinguishes Candida tropicalis (positivereaction) from Candida lusitaniae (negative reaction).

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8- TEST PERFORMANCEThe AUXACOLOR TM 2 kit is able to identify 31 yeast species and oneunicellular alga species, Prototheca wickerhamii. Compared toAUXACOLOR™, this kit can identify 7 new species of yeasts: Candidaciferrii, Candida dubliniensis, Candida sake, Kloeckera apiculata,Trichosporon asahii, Trichosporon inkin, Trichosporon mucoides.A prospective clinical study and a retrospective study on 120 strainsrepresentative of the species identified by the AUXACOLOR™ 2 kit,showed that:- 92.5% of strains tested were identified within 48 h (59.2% in 24 h),

due to the use of new tests in the ki t (hexosaminidase andprolinearylamidase tests), but also due to completion of the list ofnumerical profiles for species listed in the instructions.

- The AUXACOLOR™ 2 kit is able to distinguish all strains of Candidadubliniensis (5 strains tested) and Candida albicans (15 strains).

- The combination of the proline-arylamidase test and the phenoloxidasetest adds another test to the microplate that is useful to identify strainsof Candida glabrata (9 strains which are distinguished from Candidazeylanoides) or distinguish Candida krusei (6 strains) from Candidalipolytica (4 strains).

- The POX test allowed detection of the phenoloxidase activity ofCryptococcus neoformans for the 4 strains tested.

9- LIMITS OF THE TEST1- The AUXACOLOR TM 2 kit only allows identification of the species of

yeasts indicated in the interpretation table.2- For the referenced species, the database only lists the numerical

profiles most frequently encountered. A numerical profile not listed maycorrespond to a rare profile of a referenced species or to a non-referenced species. Refer to the interpretation table to complete theinformation contained in the database.

12

3- Exceptionally, rare species, not referenced in the instructions, may havean identical numerical profile to that of a referenced species.

4- The AUXACOLOR TM 2 microplate must only be used with pure strains.Combinations of yeasts are detected in 16% of samples. Culture onCandiSelect™ 4 or PCB medium performed in parallel with theAUXACOLOR™ 2 test makes it possible to visualise combinations ofyeasts.

5- In some cases, the isolation medium used can influence the colourchange of certain tests at 24 h (xylose, hexosaminidase) withoutinterfering with identification of the yeasts at 48 h.

6- The phenoloxidase/proline-arylamidase test:- rare strains of Cryptococcus neoformans may not express the

phenoloxidase character.- a few strains of Geotrichum capitatum, Geotrichum candidum,

Trichosporon spp and Cryptococcus laurentii can induce a chestnutgrey colour of the well. In this case the reaction is negative andwhen scoring the value "zero" must be given to this well.

- some strains of Prototheca wickerhamii may express the proline-arylamidase character.

7- Instances of reversion from "positive" to "negative", i.e. the change ofcolour of a well initially interpreted as "positive" to "negative", shouldnot be taken into account. For example, a well which is yellow at 24and 48 hours and which becomes green or blue at 48 or 72 hours isconsidered as positive.

8- On the other hand, any change from "negative" to "positive" duringthe first 72 hours must be taken into account: the well concernedshould be considered as positive. For example, a blue-grey or blue-green well at 24 or 48 hours which becomes yellow at 48 or 72 hoursis considered as positive.

10-QUALITY CONTROLAUXACOLOR TM 2 microplates (R1) are submitted to quality control usinga panel of pure strains with well defined biochemical characters.

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A control of the activity of the kit can also be performed in the laboratoryusing the following reference strains:

Characters observed after 48 h of incubation for strains cultured onSabouraud agar.

ATCC: American Type Culture Collection, 12301 Parklawn Drive,Rockville, Maryland 20852, USA.BCCM / IHEM: Belgian Co-ordinated Collections of Micro-organisms /IHEM Culture Collection, Scientific Institute of Public Health - Louis Pasteur,Mycology Section, Rue J. Wytsmanstraat 14, B-1050 Brussels.IP: Collection Nationale de Cultures de Microorganismes, Institut Pasteur,25 rue du Docteur Roux, 75724 Paris cedex 15, France.

11-MANUFACTURER'S QUALITY CONTROLAll manufactured reagents are prepared according to our Quality System,starting from reception of raw material to the final commercialization ofthe product.Each lot is submitted to quality control assessments and is only released tothe market, after conforming to pre-defined acceptance criteria.The records relating to production and control of each single lot are keptwithin Bio-Rad.

1S

14

STRAINS NEG. GLU. MAL. SAC. GAL. LAC. RAF. INO. CEL. TRE. ADO. MEL. XYL. ARA. HEX. POX./PRO.

C. NEOFORMANS - + + + + - - + V V - V V V - + -ATCC 32045T. MUCOIDES - + + + + + + + + + + + + + + - -BCCM/IHEM 14146C. LIPOLYTICA - + - - - - - - - - - - - - - - +IP 817.63

15

Code10000 -(+) 05 Candida krusei10000 - 25 Geotrichum capitatum10000 +(-) 01 Candida zeylanoides10000 +(-) 05 Candida lipolytica10000 + 05 Candida norvegensis10000 v 04 Candida inconspicua10010 +(-) 01 Candida zeylanoides10010 +(-) 05 Candida lipolytica10040 -(+) 21 Geotrichum candidum10200 - 00 Kloeckera apiculata10200 + 05 Candida norvegensis10200 +(-) 05 Candida lipolytica10240 + 05 Candida norvegensis10400 - 04 Candida glabrata10400 +(-) 01 Candida zeylanoides10402 +(-) 01 Candida zeylanoides10410 +(-) 01 Candida zeylanoides10600 +(-) 01 Candida zeylanoides10610 +(-) 01 Candida zeylanoides

33242 - 25 Trichosporon asahii33243 - 25 Trichosporon asahii33262 - 25 Trichosporon asahii33263 - 25 Trichosporon asahii33342 - 25 Trichosporon asahii33343 - 25 Trichosporon asahii33362 - 25 Trichosporon asahii33363 - 25 Trichosporon asahii33642 - 25 Trichosporon asahii33643 - 25 Trichosporon asahii33662 - 25 Trichosporon asahii33663 - 25 Trichosporon asahii33742 - 25 Trichosporon asahii33743 - 25 Trichosporon asahii33762 - 25 Trichosporon asahii11000 - 25 Geotrichum capitatum

11000 +(-) 05 Candida lipolytica11000 v 05 Candida rugosa11001 v 05 Candida rugosa11010 +(-) 05 Candida lipolytica11010 v 05 Candida rugosa11040 v 05 Candida rugosa11040 -(+) 21 Geotrichum candidum11041 v 05 Candida rugosa11050 v 05 Candida rugosa11050 -(+) 21 Geotrichum candidum11400 - 04 Prototheca wickerhamii11400 +(-) 01 Candida zeylanoides11410 +(-) 01 Candida zeylanoides11600 +(-) 01 Candida zeylanoides11610 +(-) 01 Candida zeylanoides

12400 - 04 Candida glabrata

13400 - 04 Prototheca wickerhamii

55000 - 05 Candida kefyr55001 - 05 Candida kefyr55040 - 05 Candida kefyr55041 - 05 Candida kefyr

Code31052 - 07 Candida albicans 231402 - 07 Candida albicans 231442 - 07 Candida albicans 231452 - 07 Candida albicans 231472 - 07 Candida albicans 2

57000 - 05 Candida kefyr57001 - 05 Candida kefyr57010 - 05 Candida kefyr57011 - 05 Candida kefyr57040 - 05 Candida kefyr57041 - 05 Candida kefyr57050 - 05 Candida kefyr57051 - 05 Candida kefyr57200 - 05 Candida kefyr57201 - 05 Candida kefyr57240 - 05 Candida kefyr57241 - 05 Candida kefyr57400 - 05 Candida kefyr57401 - 05 Candida kefyr57440 - 05 Candida kefyr57441 - 05 Candida kefyr

31002 - 07 Candida albicans 231042 - 07 Candida albicans 2

16

Code57600 - 05 Candida kefyr57601 - 05 Candida kefyr57640 - 05 Candida kefyr57641 - 05 Candida kefyr

71002 + 07 Candida dubliniensis71010 + 07 Candida dubliniensis71012 + 07 Candida dubliniensis71021 v 05 Candida parapsilosis71031 v 05 Candida parapsilosis71042 + 07 Candida albicans171052 + 07 Candida albicans 171061 v 05 Candida parapsilosis71071 v 05 Candida parapsilosis71124 - 44 Cryptococcus neoformans71125 - 44 Cryptococcus neoformans71134 - 44 Cryptococcus neoformans71135 - 44 Cryptococcus neoformans71164 - 44 Cryptococcus neoformans71165 - 44 Cryptococcus neoformans71174 - 44 Cryptococcus neoformans71175 - 44 Cryptococcus neoformans71324 - 44 Cryptococcus neoformans

Code71325 - 44 Cryptococcus neoformans71334 - 44 Cryptococcus neoformans71335 - 44 Cryptococcus neoformans71400 - 04 Saccharomyces cerevisiae71402 + 07 Candida dubliniensis71410 + 07 Candida dubliniensis71411 v 05 Candida parapsilosis71412 + 07 Candida dubliniensis71420 - 04 Saccharomyces cerevisiae71421 v 05 Candida parapsilosis71422 v 01 Candida sake71430 + 07 Candida dubliniensis71431 v 05 Candida parapsilosis71432 v 01 Candida sake71432 + 07 Candida dubliniensis71440 + 07 Candida albicans 171441 + 07 Candida albicans 171442 + 07 Candida albicans 171443 + 07 Candida albicans 171450 + 07 Candida albicans 171451 + 07 Candida albicans 171451 v 05 Candida parapsilosis71452 + 07 Candida albicans 171453 + 07 Candida albicans 171460 -(+) 05 Candida tropicalis71461 v 05 Candida parapsilosis71462 v 01 Candida sake71462 + 07 Candida albicans 171470 -(+) 05 Candida tropicalis71471 v 05 Candida parapsilosis71472 v 01 Candida sake71472 + 07 Candida albicans 171473 + 07 Candida albicans 171524 - 44 Cryptococcus neoformans71525 - 44 Cryptococcus neoformans71534 - 44 Cryptococcus neoformans71535 - 44 Cryptococcus neoformans71553 - 05 Candida ciferrii71560 v 40 Cryptococcus uniguttulatus71561 v 40 Cryptococcus uniguttulatus71564 - 44 Cryptococcus neoformans

70160 v 40 Cryptococcus uniguttulatus70161 v 40 Cryptococcus uniguttulatus70170 v 40 Cryptococcus uniguttulatus70171 v 40 Cryptococcus uniguttulatus70400 - 04 Saccharomyces cerevisiae70462 v 01 Candida sake70560 v 40 Cryptococcus uniguttulatus70561 v 40 Cryptococcus uniguttulatus70570 v 40 Cryptococcus uniguttulatus70571 v 40 Cryptococcus uniguttulatus70620 v 05 Candida lusitaniae70660 v 40 Cryptococcus albidus70661 v 40 Cryptococcus albidus70662 v 01 Candida sake70670 v 05 Candida lusitaniae70760 v 40 Cryptococcus albidus70761 v 40 Cryptococcus albidus

17

Code71565 - 44 Cryptococcus neoformans71570 v 40 Cryptococcus uniguttulatus71571 v 40 Cryptococcus uniguttulatus71574 - 44 Cryptococcus neoformans71575 - 44 Cryptococcus neoformans71620 v 05 Candida lusitaniae71620 v 40 Cryptococcus albidus71630 v 05 Candida lusitaniae71660 -(+) 05 Candida tropicalis71660 v 05 Candida lusitaniae71661 v 05 Candida lusitaniae71662 v 01 Candida sake71670 -(+) 05 Candida tropicalis71670 v 05 Candida lusitaniae71670 v 40 Cryptococcus albidus71671 + 05 Candida guilliermondii71671 v 05 Candida lusitaniae71672 v 01 Candida sake71724 - 44 Cryptococcus neoformans71725 - 44 Cryptococcus neoformans71734 - 44 Cryptococcus neoformans71735 - 44 Cryptococcus neoformans71764 - 44 Cryptococcus neoformans71765 - 44 Cryptococcus neoformans71774 - 44 Cryptococcus neoformans71775 - 44 Cryptococcus neoformans

72762 - 25 Trichosporon inkin72763 - 25 Trichosporon inkin

Code73653 -(+) 21 Trichosporon spp73661 -(+) 21 Trichosporon spp73662 -(+) 25 Trichosporon spp73663 - 25 Trichosporon asahii73670 v 05 Candida lusitaniae73671 -(+) 21 Trichosporon spp73673 -(+) 21 Trichosporon spp73741 -(+) 21 Trichosporon spp73742 - 25 Trichosporon asahii73743 - 25 Trichosporon asahii73751 -(+) 21 Trichosporon spp73753 -(+) 21 Trichosporon spp73761 -(+) 21 Trichosporon spp73762 -(+) 25 Trichosporon spp73763 -(+) 25 Trichosporon spp73771 -(+) 21 Trichosporon spp73773 -(+) 21 Trichosporon spp

74000 - 04 Saccharomyces cerevisiae74020 - 04 Saccharomyces cerevisiae74361 v 40 Cryptococcus albidus74371 v 40 Cryptococcus albidus74400 - 04 Saccharomyces cerevisiae74420 - 04 Saccharomyces cerevisiae74560 v 40 Cryptococcus uniguttulatus74561 v 40 Cryptococcus uniguttulatus74570 v 40 Cryptococcus uniguttulatus74571 v 40 Cryptococcus uniguttulatus74660 v 40 Cryptococcus albidus74661 v 40 Cryptococcus albidus74760 v 40 Cryptococcus albidus74761 v 40 Cryptococcus albidus74771 v 40 Cryptococcus albidus

73242 - 25 Trichosporon asahii73243 - 25 Trichosporon asahii73262 - 25 Trichosporon asahii73263 - 25 Trichosporon asahii73342 - 25 Trichosporon asahii73343 - 25 Trichosporon asahii73362 - 25 Trichosporon asahii73363 - 25 Trichosporon asahii73641 -(+) 21 Trichosporon spp73642 -(+) 25 Trichosporon spp73643 - 25 Trichosporon asahii73651 -(+) 21 Trichosporon spp

75000 - 04 Saccharomyces cerevisiae75020 - 04 Saccharomyces cerevisiae75124 - 44 Cryptococcus neoformans75125 - 44 Cryptococcus neoformans75134 - 44 Cryptococcus neoformans75135 - 44 Cryptococcus neoformans75164 - 44 Cryptococcus neoformans75165 - 44 Cryptococcus neoformans

18

Code75774 - 44 Cryptococcus neoformans75775 - 44 Cryptococcus neoformans

76361 v 40 Cryptococcus albidus76371 v 40 Cryptococcus albidus76761 v 40 Cryptococcus albidus76771 v 40 Cryptococcus albidus

Code75174 - 44 Cryptococcus neoformans75175 - 44 Cryptococcus neoformans75324 - 44 Cryptococcus neoformans75325 - 44 Cryptococcus neoformans75334 - 44 Cryptococcus neoformans75335 - 44 Cryptococcus neoformans75361 v 40 Cryptococcus albidus75371 v 40 Cryptococcus albidus75400 - 04 Saccharomyces cerevisiae75420 - 04 Saccharomyces cerevisiae75524 - 44 Cryptococcus neoformans75525 - 44 Cryptococcus neoformans75534 - 44 Cryptococcus neoformans75535 - 44 Cryptococcus neoformans75551 - 05 Candida ciferrii75553 - 05 Candida ciferrii75560 v 40 Cryptococcus uniguttulatus75561 v 40 Cryptococcus uniguttulatus75564 - 44 Cryptococcus neoformans75565 - 44 Cryptococcus neoformans75570 v 40 Cryptococcus uniguttulatus75571 v 40 Cryptococcus uniguttulatus75574 - 44 Cryptococcus neoformans75575 - 44 Cryptococcus neoformans75630 v 00 Candida famata75631 + 05 Candida guilliermondii75631 v 00 Candida famata75651 v 00 Candida famata75670 v 00 Candida famata75671 + 05 Candida guilliermondii75671 v 00 Candida famata75724 - 44 Cryptococcus neoformans75725 - 44 Cryptococcus neoformans75734 - 44 Cryptococcus neoformans75735 - 44 Cryptococcus neoformans75751 - 05 Candida ciferrii75753 - 05 Candida ciferrii75761 v 40 Cryptococcus albidus75764 - 44 Cryptococcus neoformans75765 - 44 Cryptococcus neoformans75771 v 40 Cryptococcus albidus

77361 v 40 Cryptococcus albidus77371 v 40 Cryptococcus albidus77610 v 00 Candida famata77611 v 00 Candida famata77630 v 00 Candida famata77631 v 00 Candida famata77641 -(+) 21 Trichosporon spp77643 -(+) 21 Trichosporon spp77651 -(+) 21 Trichosporon spp77651 v 00 Candida famata77653 -(+) 21 Trichosporon spp77661 -(+) 21 Trichosporon spp77663 -(+) 21 Trichosporon spp77670 v 00 Candida famata77671 -(+) 21 Trichosporon spp77671 v 00 Candida famata77673 -(+) 21 Trichosporon spp77741 -(+) 21 Trichosporon spp77742 -(+) 21 Trichosporon spp77743 -(+) 21 Trichosporon spp77751 -(+) 21 Trichosporon spp77753 -(+) 21 Trichosporon spp77761 -(+) 21 Trichosporon spp77761 v 40 Cryptococcus albidus77763 - 25 Trichosporon mucoides77771 - 25 Trichosporon mucoides77771 - 40 Cryptococcus laurentii77771 v 40 Cryptococcus albidus77772 -(+) 21 Trichosporon spp77773 - 25 Trichosporon mucoides

12-BIBLIOGRAPHY1. C.P. KURTZMAN, J.W. FELL

The Yeasts. A taxonomic study, Fourth Revised and Enlarged Edition.Editions Elsevier 1998.

2. BH COOPER, M. SILVA-HUNTERYeasts of medical importance. Manual of clinical microbiology. Fourth edition. 1985, p 526-541.

3. H. KOENIGGuide de mycologie médicale. Editions Ellipses 1995, p 31-96.

4. H. FRICKER-HIDALGO, B. LEBEAU, P. KERVROEDAN, O. FAURE, P. AMBROISE-THOMAS, R. GRILLOTAUXACOLOR TM, a new commercial system for yeast identification:evaluation of 182 strains comparatively with ID 32C. Ann. Biol. Clin. (Paris). 1995; 53 (4): 221-5.

5. H. FRICKER, D. MONGET, B. LEBEAU, M. BABOLAT, P. AMBROISE-THOMAS, R. GRILLOTRapid identification of Candida albicans: evaluation of “Rapidecalbicans". Study of 444 yeast strains. Ann. Biol. Clin. (Paris). 1992; 50 (2): 103-6.

6. D. SULLIVAN , D. COLEMANCandida dubliniensis : characteristics and identification.J. Clin. Microbiol. 1998 ; 36 (2) : 329-334.

7. H.S.WANG, R.T. ZEIMIS, G.D. ROBERTSEvaluation of a caffeic acid-ferric citrate test for rapid identification ofCryptococcus neoformans. J. Clin. Microbiol. 1977; 6 (5): 445-9.

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