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Journal of Clinical Microbiology, April 2001, p. 1647-1649, Vol. 39, No. 4
0095-1137/01/$04.00+0 DOI: 10.1128/JCM.39.4.1647-1649.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.
Evaluation of Candida ID, a New Chromogenic Medium
for Fungal Isolation and Preliminary Identification of Some
Yeast Species
H.
Fricker-Hidalgo,1,*
S.
Orenga,2
B.
Lebeau,1
H.
Pelloux,1
M. P.
Brenier-Pinchart,1
P.
Ambroise-Thomas,1 and
R.
Grillot1
Service de Parasitologie-Mycologie, Centre
Hospitalier Universitaire, 38043 Grenoble,1 and
bioMérieux, Le Zaligniez, 38390 La
Balme-les-Grottes,2 France
Received 3 October 2000/Returned for modification 14 November
2000/Accepted 19 January 2001
 |
ABSTRACT |
Candida ID, a new chromogenic medium, allows identification of
Candida albicans (blue colonies) and preliminary
identification into a group of four species (pink colonies). In
comparison with Albicans ID2 and Sabouraud gentamicin chloramphenicol
on 446 fungal strains, Candida ID allowed the isolation of more species
than Albicans ID 2 (95.5% versus 91.2%).
| |
TEXT |
Mycoses are a growing medical
problem requiring prompt diagnosis with species identification and
early adapted antifungal therapy (3, 7, 11, 14). The
importance of identifying the pathogen quickly (20), as
well as the difficulty in detecting mixed cultures in the same plate
with the traditional Sabouraud glucose agar medium (2),
has fostered the development of differential media for the
identification of yeasts (1, 5, 6, 8, 12, 18, 19, 22).
Candida ID (CAID) (bioMérieux, Marcy l'Etoile, France) is a
recently commercialized, ready-to-use medium. CAID was developed to
improve the isolation of yeasts, the identification of Candida
albicans and C. dubliniensis, the detection of mixed cultures, and the preliminary identification of other species (C. tropicalis, C. guilliermondii, C. kefyr, and C. lusitaniae, which produce pink colonies). Further testing has to
be performed to separate these four species. In our study, CAID was
compared with Albicans ID2 (AID2) (bioMérieux) and Sabouraud
gentamicin chloramphenicol agar (SGC) using 307 collection strains and
139 strains from 220 clinical samples.
A total of 307 fungal strains (247 yeasts and 60 filamentous fungi)
from bioMérieux and Grenoble hospital collections were tested on
the three media. The collection strains were subcultured on Sabouraud
agar plates and incubated for 2 to 5 days at 35 or 27°C according to
the species. For all isolates, a suspension of the organisms was made
in physiological saline and seeded onto three plates (CAID, AID2, and
SGC). The plates were incubated at 35°C except for
Penicillium and Fusarium spp., which were
incubated at 27°C. Equivalent amounts of each of the 220 clinical
samples were directly applied to the three media and incubated at
35°C. The reading of the plates and interpretation of the results
were carried out after 24, 48, and 72 h of incubation. The
determination of the color, the number, and the diameter of the
different colonies grown on CAID, AID2, and SGC was performed at each
reading step. The colonies showing different morphologies on media
inoculated with clinical samples were picked up and identified by
conventional mycological methods (15): Bichrolatex
albicans, Krusei color (Fumouze, Levallois-Perret, France)
(13), ID32C (bioMérieux), and macroscopic and
microscopic observations. The colony detections on CAID, AID2, and SGC
were compared and analyzed in terms of sensitivity (number of true
positives/number of true positives plus the number of false negatives)
and specificity (number of true negatives/number of true negatives plus
the number of false positives). The data were statistically analyzed by
2 test. C. albicans identification on CAID
and AID2 was analyzed in terms of sensitivity and specificity using the
described conventional mycological methods as a reference.
CAID supported the growth of more strains (426 of 446, 95.5%) than
AID2 (407 of 446, 91.2%) and as many as SGC (426 of 446, 95.5%)
(Tables 1 and
2). The differences between CAID and AID2 were statistically significant (P < 0.025). Among the
220 clinical specimens, 103 gave positive cultures (43 stools, 14 tracheal fluids, 14 bronchoscopic aspirations, 30 sputum specimens, and 2 miscellaneous samples such as drain fluid) (Table 2). For the 31 polyfungal specimens, AID2 and CAID supported the growth of more fungus
strains (62 of 67, 92.5%) than SGC (53 of 67, 79.1%), with
statistically significant differences (P < 0.05). The
sensitivities of CAID and AID2 for detection and differentiation of
C. albicans were 97.6% (125 of 128) and 100% (128 of 128),
respectively, and the specificities were 89.7 and 82.7%, respectively.
The colonies of 17 among 36 isolates of C. tropicalis were
blue on AID2 but not on CAID. CAID and AID2 did not enable
differentiation between C. albicans and C. dubliniensis. On CAID, colonies of C. tropicalis (32 of
36), C. lusitaniae (10 of 10), C. guilliermondii
(9 of 10), and C. kefyr (14 of 14) were pink after 72 h
of incubation and needed complete identification. The time to detection
of fungi growing on all three media (388 strains) were determined.
After 24 h of incubation at 35°C, 90.7% (352 of 388) of the
fungus strains were detected on SGC, 86.3% (335 of 388) were detected
on AID2, and 89.7% (348 of 388) were detected on CAID. The average
sizes of the colonies of collection strains after 24 h of
incubation on SGC, AID2, and CAID were 0.63 ± 0.46, 0.86 ± 0.56, and 0.94 ± 0.55 mm, respectively. Bacteria were isolated
from 23 of the 220 clinical samples (10.5%) on at least one of the
three media. The numbers of bacterial strains isolated on SGC, AID2,
and CAID were 14, 14, and 20, respectively.
View this table:
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TABLE 1.
Colonial coloration observed after 72 h of
incubation on the three media. (SGC, AID2, and CAID) inoculated with
307 collection strains
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View this table:
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[in a new window]
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TABLE 2.
Colonial coloration observed after 72 h of
incubation on the three media (SGC, AID2, and CAID) inoculated with 103 clinical samples
|
|
CAID was developed to obtain better results than with AID2 (4, 9,
10, 17, 21). Our study showed that CAID allowed a better
detection of different types of collection strains than AID2 (95.4 and
89.2%, respectively) and that the results from clinical samples were
equivalent with CAID and AID2 (95.7%). The collection strains detected
only on CAID belonged to species that were not isolated from the
clinical samples in our study (C. colliculosa, C. famata, C. intermedia, C. sphaerica, C. utilis, C. zeylanoides, Cryptococcus
humicolus, Cryptococcus laurentii, and Rhodotorula rubra). The other advantage of CAID related to the specificity of
C. albicans identification. No C. tropicalis
colonies were blue on CAID in contrast to what happens on AID2.
C. lodderae and C. rugosa, which produce blue
colonies on CAID, were rarely isolated from clinical specimens
(16), blue Trichosporon colonies were easily
differentiated from C. albicans by their macroscopic morphology, while C. dublinensis was not distinguished from
C. albicans. Furthermore, when different yeast species were
mixed in a single sample and plated out on CAID, the distinction
between colony colors and forms was extremely easy to do.
Discrimination of C. kefyr, C. lusitaniae, C. tropicalis,
and C. guilliermondii from other yeasts by their pink
colonies facilitates their definitive identification particularly for
C. guilliermondii which has the same profile as C. famata on the ID32C strip. On CAID, the colonies of C. guilliermondii were pink while the colonies of C. famata were white; thus, no other complementary test (different
from ID32C) was necessary.
In conclusion, our study shows that CAID is an effective, ready-to-use
medium for the isolation of fungi and identification of C. albicans from samples in clinical laboratories.
| |
ACKNOWLEDGMENTS |
We thank L. Barbaux, C. Cotte, A. Rongier, L. Coeur, C. Pinel, and
A. De Bengy for their technical assistance and S. Bastin for rereading
the manuscript.
| |
FOOTNOTES |
*
Corresponding author. Mailing address: Service de
Parasitologie-Mycologie, Centre Hospitalier Universitaire, B.P. 217, 38043 Grenoble, France. Phone: 33-4-76-76-54-90. Fax: 33-4-76-76-56-60. E-mail: HFricker-Hidalgo{at}chu-grenoble.fr.
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Journal of Clinical Microbiology, April 2001, p. 1647-1649, Vol. 39, No. 4
0095-1137/01/$04.00+0 DOI: 10.1128/JCM.39.4.1647-1649.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.
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