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Journal of Clinical Microbiology, May 1998, p. 1443-1445, Vol. 36, No. 5
0095-1137/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.
Comparison of the Rapid Yeast Plus Panel with the
API20C Yeast System for Identification of Clinically Significant
Isolates of Candida Species
Judith S.
Heelan,1,2,*
Edgar
Sotomayor,1
Kimberly
Coon,1 and
Julia B.
D'Arezzo3
Department of Pathology, Memorial Hospital of Rhode Island,
Pawtucket, Rhode Island 028601;
Department of Pathology, Veterans' Administration Medical
Center, Providence, Rhode Island 029083; and
Department of Pathology and Laboratory Medicine, Brown
University School of Medicine, Providence, Rhode Island
029012
Received 30 December 1997/Returned for modification 26 January
1998/Accepted 3 February 1998
 |
ABSTRACT |
The RapID Yeast Plus system (Innovative Diagnostic Systems,
Norcross, Ga.) is a qualitative micromethod employing conventional tests and single-substrate chromogenic tests and having a 4-h incubation period. This system was compared with the API20C (bioMerieux Vitek, Hazelwood, Mo.) system, a 24- to 72-h carbohydrate assimilation method. One hundred thirty-three clinical yeast isolates, including 57 of Candida albicans, 26 of Candida tropicalis,
23 of Candida glabrata, and 27 of other yeasts, were tested
by both methods. When discrepancies occurred, isolates were further
tested by the Automated Yeast Biochemical Card (bioMerieux Vitek). Germ
tube production and microscopic morphology were used as needed to
definitively identify yeast isolates. The RapID Yeast Plus system
correctly identified 125 yeast isolates, with an overall accuracy of
94% (125 of 133). Excellent correlation was found in the recognition of the three yeasts most commonly isolated from human sources. The test
was 99% (105 of 106 isolates) accurate with C. albicans, C. tropicalis, and C. glabrata. The RapID Yeast
Plus system compares favorably with the API20C system and provides a
simple, accurate alternative to conventional assimilation methods for
the rapid identification of the most commonly encountered isolates of
Candida species.
 |
TEXT |
Recently, there has been an increase
in fungal infections in immunocompetent as well as immunocompromised
patients. Yeasts (particularly Candida albicans) are the
most common fungi isolated from human infections. Various yeast species
are inherently or potentially resistant to amphotericin B and the new
azole agents (13, 14). Rapid identification of significant
clinical yeast isolates is therefore imperative for prompt institution
of appropriate antifungal therapy. Conventional methods for the
definitive identification of clinically significant yeasts to species
level are generally based upon the ability of the yeast to utilize
sugars by fermentation or assimilation (9, 14). The API20C
(20C) system consists of strips containing dehydrated substrates for
carbohydrate assimilation reactions. Although the test is simple and
accurate, results are not available for 24 to 72 h. Recently, the
RapID Yeast Plus (RYP) panel, which requires only a 4-h incubation
period, was introduced. This system of biochemical reactions, based
upon single-substrate enzymatic test reactions involving preformed
enzymes in the inoculum, greatly decreases the turnaround time for the
identification of the most common yeasts isolated from human patients.
The purpose of this study was to evaluate the performance of this rapid
test compared to that of the older, slower API system.
A total of 133 clinical yeast isolates were examined in this study.
Specimens were obtained from inpatients and outpatients seen in
satellite clinics affiliated with Memorial Hospital of Rhode Island.
All yeasts were tested with the RYP panel and the 20C test strip.
Discrepancies were resolved with the Vitek (VIT) Yeast Biochemical Card
and germ tube production, as well as microscopic morphologic
examination for blastoconidia, chlamydospores, pseudohyphae, and true
hyphae, following growth on cornmeal Tween 80 agar, after incubation at
25°C for 72 h. The 20C yeast strip was inoculated with each
yeast suspended in basal medium, which was heated before use, following
the manufacturer's instructions. Following incubation at 30°C,
growth in each well was recorded at 24, 48, and 72 h. Final
identification was made with the Analytical Profile index, when the
selection was described as excellent, very good, or acceptable. The RYP
was inoculated with a yeast suspended in RapID inoculation fluid,
according to the manufacturer's instructions. Following incubation at
30°C for 4 h, color development was noted in each test cavity
either directly or after adding reagents. A minimal microcode was
constructed and interpreted by comparison of test results to reactivity
patterns stored in a database.
The VIT Yeast Biochemical Card is designed for use with the automated
Vitek system. Each card was inoculated with a yeast isolate suspended
in 0.45% NaCl to a McFarland no. 2 standard, according to the
manufacturer's instructions. Suspensions were inoculated into the card
with the filling module, sealed with the sealing module, and incubated
at 30°C for 24 or 48 h. The cards were read by the reader
module; biochemical reactions were analyzed and interpreted by matching
biocodes with the database of the computer. Germ tube production was
determined microscopically for all isolates after incubation of the
yeast at 37°C in rabbit plasma for 2.5 h.
All isolates of C. albicans produced germ tubes; all
non-C. albicans isolates were germ tube negative. Eighty
percent of the yeasts isolated were identified as either C. albicans (43% [57 of 133]), Candida tropicalis (20%
[26 of 133]), or Candida glabrata (27% [23 of 133]).
The accuracy of the RYP with this commonly isolated group of yeasts was
99% (105 of 106). Of this group, one isolate of C. tropicalis was misidentified as Candida stellatoidea by
RYP (Table 1). There was initial
agreement between the two methods among 92% (122 of 133) of the
isolates tested. The 11 isolates yielding discrepant results with RYP
and 20C were further studied by morphological examination and with the
VIT card. These studies confirmed results obtained with RYP in three
cases, giving the RYP an overall accuracy of 94% (125 of 133). Two of
the three isolates were identified as C. glabrata, producing
blastoconidia but no pseudohyphae on cornmeal agar. One isolate was
identified as C. tropicalis, showing robust blastoconidia
arranged in small groups along pseudohyphae and true hyphae
microscopically. These three isolates gave excellent identification
(99%) on the Vitek instrument. The eight remaining yeasts
(misidentified by RYP) were confirmed morphologically (see Table 2).
Seven of these isolates were correctly identified by the Vitek
instrument; one isolate of Candida parapsilosis was
misidentified by Vitek as C. tropicalis. Microscopic
morphology confirmed 20C results in all eight cases. Certain isolates
were problematic (Table 2). Several
yeasts consistently gave no acceptable codes; these yeasts were
Trichosporon beigelii (two isolates) and
Rhodotorula rubra (one isolate), rarely
isolated from our patient population. A cost analysis of each system
including materials and labor showed $7.85, $7.40, and $7.00 for an RYP
test, a 20C test, or a Vitek assay, respectively.
Conventional methods for the identification of clinically significant
yeasts are time-consuming and labor-intensive. Rapid screening methods,
including the ability to produce germ tubes (8, 13) and
production of the enzymes beta-galactosaminidase and
L-proline aminopeptidase (2, 4, 7, 11), may be used to presumptively identify C. albicans. Other rapid
screening methods involve the use of differential culture media
(1, 6, 10, 12). Although cellular fatty acid analysis by
gas-liquid chromatography has been successfully used to identify
mycobacteria and other fastidious microorganisms, the current database
is considered to be inadequate for the routine identification of
clinically significant yeasts (3). The 20C system was one of
the first standardized commercial products used for rapid yeast
identification and is frequently used as a reference method (5,
8). Although the VIT Yeast Biochemical Card has been reported to
be inadequate for the rapid identification of uncommon yeast isolates,
it has been shown to provide accurate identification of the most
commonly isolated yeasts (5). Although both the 20C and VIT
tests yield accurate identification of most yeast isolates from
clinical sources, a minimum of 24 and often 48 h of incubation is
required, especially for slowly growing yeasts (8). Results
of our study show that the RYP system is highly accurate in identifying
the most commonly isolated yeasts. The cost per test for the RYP system
is comparable to that of the API20C system; the RYP assay is an
accurate, user-friendly method for same-day identification of
Candida species.
 |
ACKNOWLEDGMENTS |
We thank Andrea Kubacki for her assistance in the preparation of
the manuscript.
 |
FOOTNOTES |
*
Corresponding author. Mailing address: Microbiology
Laboratory, Memorial Hospital of Rhode Island, 111 Brewster St.,
Pawtucket, RI 02860. Phone: (401) 729-2839. Fax: (401) 729-2990.
 |
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Journal of Clinical Microbiology, May 1998, p. 1443-1445, Vol. 36, No. 5
0095-1137/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.
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