Rapid PCR-based test for identifying Candida albicans by using

primers derived from the pH-regulated KER1 gene

Amparo Galán1, Verónica Veses2, Amelia Murgui3, Manuel Casanova4 & José P. Martı́nez4
1
Banco de lı́neas celulares, Centro de Investigación Prı́ncipe Felipe, Valencia, Spain; 2Molecular and Cell Biology Department, School of Medical
Sciences, University of Aberdeen, Aberdeen, UK; 3Departamento de Bioquı́mica y Biologı́a Molecular, Facultad de Farmacia, Universitat de Valencia,

Downloaded from https://academic.oup.com/femsyr/article/6/7/1094/491946 by Semenov Institite for Chemical Physics user on 24 June 2023
Burjasot, Valencia, Spain; and 4Departamento de Microbiologı́a y Ecologı́a, Facultad de Farmacia, Universitat de Valencia, Burjasot, Valencia, Spain

Correspondence: José P. Martı́nez, Abstract

Departamento de Microbiologı́a y Ecologı́a,
Facultad de Farmacia, Universitat de Valencia,
Burjasot, Valencia, Spain. Tel.: 134 96
3544770; fax: 134963544299; e-mail:
jose.pedro.martinez@uv.es
Received 26 January 2006; revised 21 April
2006; accepted 25 April 2006.
First published online 9 June 2006.
DOI:10.1111/j.1567-1364.2006.00114.x
Editor: Rafael Sentandreu
Keywords
Candida albicans ; molecular identification;
PCR; KER1 gene; DNA extraction.
A PCR-based method in combination with a simple, reliable and inexpensive DNA
extraction procedure for rapid detection of Candida albicans clinical isolates is
described here. The extraction protocol is based on a combination of chemical
(NaOH and detergents) and physical (boiling) treatments, thus avoiding many of
the problems inherent in the currently available DNA extraction protocols
(basically the use of expensive and/or toxic chemical reagents), and may be useful
for daily clinical routine. The PCR-based system described here uses a single pair of
primers (SC1F and SC1R) deduced from the C. albicans-specific KER1 gene
sequence. These primers amplify a 670-bp fragment of the KER1 gene. All the
clinical C. albicans isolates generated the expected 670-bp amplicon. Other non-
albicans Candida species, including the azole-resistant C. krusei and C. glabrata,
and the very closely related C. dubliniensis, failed to amplify any DNA fragment.
The PCR results reported here suggest that amplification with SC1F and SC1R
primers is species specific and, consequently, may be useful for specifically
identifying C. albicans strains.

Introduction known to be the most virulent of the medically important

Systemic candidiasis is a severe infectious complication that
is particularly frequent in neutropenic patients. Candidemia
has risen to become the fourth most frequent cause of
bloodstream infection, with a mortality rate of about 50%.
In this context, Candida albicans is the most common
pathogenic Candida species. Diagnosis of invasive candidia-
sis is often very difficult, since most clinical symptoms of
disseminated candidiasis are nonspecific, and blood cultures
are frequently negative or become positive too late; thus,
more than 50% of blood cultures initially reported as
negative have been shown to be from proven cases of
systemic candidiasis in a later autopsy. Consequently, there
is increasing interest in the use of molecular-based methods
for early and unequivocal diagnosis of these infections.
The incidence of systemic candidiasis has increased
steadily over the last two decades (Beck-Sagué & Jarvis,
1993; Fischer-Hoch & Hutwagner, 1995; Jarvis, 1995).
Several factors can predispose individuals to superficial
and systemic candidiasis, including immunosuppressive
therapy, prolonged broad-spectrum antibiotic therapy,
invasive devices such as catheters, and prolonged hospital
stays (Jarvis, 1995; Wenzel, 1995). Candida albicans is
Candida species (Calderone, 2002). However, some emer-
gent non-C. albicans species, including C. glabrata and C.
krusei, exhibit reduced susceptibility to commonly used
antifungal agents relative to C. albicans (Nguyen et al.,
1996; Pfaller et al., 1999), thus making it necessary for the
infected organism to be identified to the species level for
optimum therapy.
Candida dubliniensis has recently been described as an
opportunistic pathogen, and thus may be considered to be
another emerging non-albicans Candida species. Candida
dubliniensis has been linked to oral candidiasis in AIDS
patients (Sullivan et al., 1995) and has also been associated
with invasive disease. Although C. albicans and C. dubli-
niensis share phenotypic and diagnostic characteristics,
they differ in their carbohydrate assimilation profiles,
growth patterns at elevated temperatures, and intracellular
b-glucosidase activities (Coleman et al., 1997; Pinjon
et al., 1998; Salkin et al., 1998). Molecular studies have
demonstrated that C. dubliniensis possesses a very distinct
genomic organization (Gilfillan et al., 1998; Sullivan &
Coleman, 1998) from C. albicans, which allows molecular
differentiation between these two species (Kurzai et al.,
1999).

c 2006 Federation of European Microbiological Societies FEMS Yeast Res 6 (2006) 1094–1100
Published by Blackwell Publishing Ltd. All rights reserved
Rapid PCR-based test for identifying Candida albicans 1095

Recently, molecular biology-based techniques have been Table 1. Fungal strains used in this study
adapted to the identification of fungal pathogens (McCul- Lane
lough et al., 1995; Elie et al., 1998; Kanbe et al., 2002; Trost in
et al., 2004). In particular, PCR has increasingly been used Strain Origin Fig. 1
for Candida diagnosis, as it is quick, simple, specific, Candida glabrata CECT1448 1
sensitive and reliable (Mannarelli & Kurtzman, 1998; Wa- Candida guilliermondii CECT1021 2

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hyuningsih et al., 2000; Arancia et al., 2004). To adapt PCR Candida kefyr CECT1436 3
to daily clinical routine, simplification and standardization Candida krusei CECT1433 4
Candida parapsilopsis Clinical isolatew 5
of DNA extraction and PCR amplification is of major
Candida parapsilopsis Clinical isolate 6
importance.
Candida kefyr CECT1018 7
In this article, a PCR-based method for the rapid detec- Candida tropicalis CECT1005 8
tion of C. albicans isolates is described. A simpler DNA Cryptococcus neoformans CECT1078 9
extraction procedure, based on alkali, detergent and heat Kluyveromyces lactis IFO1267z 10
treatment, which was previously described for the recovery Rhodotorula marina CECT1139 11
of Candida DNA, and the use of primers derived from the Saccharomyces cerevisiae CECT1170 12
Schizosaccharomyces pombe CECT1375 13
specific C. albicans pH-regulated KER1 gene, coding for a
Yarrowia lipolytica INAG135668‰ 14
novel lysine/glutamic acid-rich protein with no significant
Candida glabrata Clinical isolate 15
homology to any other known sequences (Galán et al., Candida albicans CECT1675 16
2004), allows C. albicans detection, identification, and Candida albicans SC5314z –
differentiation from all non-albicans Candida species, in- Candida dubliniensis Clinical isolate –
cluding the azole-resistant C. krusei and C. glabrata, and the Colección Española de Cultivos Tipo, Burjasot, Valencia, Spain.
phenotypically related C. dubliniensis, all of which are of w
Servicio de Microbiologı́a, Hospital Universitario la Fe, Valencia, Spain;
clinical and epidemiologic relevance. identity of clinical isolates assayed in this work belonging to the genus
Candida was further checked when required by means of commercial
Materials and methods biochemical and serological tests (see Materials and methods section).
z
Institute of Fermentation, Osaka, Japan.
‰
Institute Nacional Agronomique Paris-Grignon, France.
Organisms and growth conditions z
See reference Gillum et al. (1984).
Fungal strains used in this work are listed in Table 1. Clinical
isolates, of both superficial and systemic origin, were
isolation. The first approach was based on Einsele’s proce-
provided by the Servicio de Microbiologia, Hospital Uni-
dure (Einsele et al., 1997), with several modifications.
versitario la Fe, Valencia, Spain. The C. albicans reference
Briefly, fungal cells were added to 1 mL of leukocyte lysis
strains used in this work as positive controls were CECT
buffer (WCLB) consisting of 10 mM Tris (pH 7.6), 10 mM
1675 and SC5314 (Gillum et al., 1984). Ura1 and Ura null
EDTA (pH 8.0), 50 mM NaCl, 0.2% sodium dodecyl sulfate
mutant strains for KER1 (C1N7 and CAC1; Galán et al.,
(SDS), and 200 mg of proteinase K (Sigma) per mL, and
2004) were used as negative controls throughout this study.
incubated at 65 1C for 1 h (A). In the second extraction
Fungal species were routinely grown on Sabouraud dextrose
procedure, fungal cells were added to 1 mL of WCLB
agar plates (Pronadisa), and YPD solid medium (1% yeast
supplemented with 300 mg of Zymolyase (ICN Biomedicals)
extract, 2% peptone, 2% glucose, 1.5% agar). CHROMagar
and 28 mM b-mercaptoethanol (Sigma), and incubated at
Candida plates (Biocheck) were used for presumptive iden-
37 1C for 45 min (B). The third extraction method involved
tification of Candida species. Definitive identification of all
the addition of the fungal cells to the same suspension as
clinical isolates assayed in this work belonging to the genus
above and incubation at 37 1C for 45 min and at 95 1C for
Candida was performed by means of biochemical methods
5 min (C). In all three cases, cells were pelleted for 5 min at
(using the API 20C AUX system), and confirmed when
5000 g, and 10 mL of the supernatant was used for the PCR
necessary with the Bichrolatex albicans agglutination test
reaction.
(Fumouze, Levallois-Perret, France).
A mechanical disruption method modified from that of
Hee Shin et al. (1997) was also used to extract fungal DNA.
Extraction of fungal DNA
Fresh yeast cells were resuspended in 300 mL of TXTE buffer
Six different fungal DNA extraction methods were tested consisting of 10 mM Tris, 1 mM EDTA and 1% Triton X-
for further comparative analysis. The method giving the 100, final pH 8.0, and transferred to a new Eppendorf tube
best result in the PCR tests was adapted for the DNA containing 200 mL of 425–600-mm glass beads (Sigma). After
extraction of all clinical isolates. In all cases, fungal isolates boiling in a water bath for 15 min, the mixture was vortexed
were cultured on solid YPD at 30 1C for 48 h prior to DNA for 15 min until complete cell breakage. The tubes were then

FEMS Yeast Res 6 (2006) 1094–1100

c2006 Federation of European Microbiological Societies
Published by Blackwell Publishing Ltd. All rights reserved
1096
centrifuged at 10 000 g for 1 min, and the supernatant was
stored at  20 1C until it was used for PCR amplification
(D). PCR amplifications were also performed directly on
intact cells without any DNA purification steps (Maneu
et al., 2000) (E).
Finally, a heat–alkali treatment for fungal DNA purifica-
tion was applied. Isolated fresh colonies were resuspended in
100 mL of 10 mM NaOH containing 0.5% Tween-20 and
0.5% Nonidet P-40. Cells were then boiled in a water bath
for 10 min and centrifuged at 10 000 g for 15 min. Ten
microliters from the supernatant was used for each PCR
reaction (F). To check the sensitivity of this method,
extractions were performed with different cell suspensions
containing different numbers of cells (from 105 to 10 mL1),
previously assessed by microscopy cell counting in a hemo-
cytometer chamber.
For Southern blot analysis, the DNA isolation method
was adapted from Hoffman and Winston (Hoffman &
Winston, 1987). Briefly, cells were grown to late stationary
growth phase in 10 mL of YPD and collected by centrifuga-
tion, and the cell pellets were resuspended in 0.2 mL of
breaking buffer (2% Triton X-100, 1% SDS, 100 mM NaCl,
10 mM Tris buffer, pH 8.0, 1 mM EDTA), and 0.2 mL of
phenol/chloroform/isoamyl alcohol (25 : 24 : 1), and 0.2 g of
glass beads was added. This mixture was vortexed, and
0.2 mL of Tris-EDTA (TE) was added. After centrifugation,
the aqueous phase was transferred to a new Eppendorf tube,
and 1 mL of ethanol was added to precipitate the DNA. After
centrifugation, the pellet was resuspended in 0.4 mL of TE
buffer and treated with 100 mg of RNase A. DNA was
precipitated again with 0.3 M (final concentration) NaCl,
and two volumes of ethanol. The DNA was finally resus-
pended in 50 mL of TE. Samples were stored at 4 1C.
Primers and PCR amplification
The oligonucleotide primer pair (SC1F, 5 0 -CGGAGATTTTCT
CAATAAGGACCAC, and SC1R, 5 0 -AGTCAATCTCTGTC-
TCCCCTTGC) was designed using information from gene
databases and our own sequence for the KER1 gene. These
primers amplify a 670-bp fragment in the KER1 gene of C.
albicans. The same batch of primers was used throughout the
study.
Amplification reactions were performed in 50-mL aliquots
of a solution containing 5 mL of 10  PCR buffer, 3.5 mM
MgCl2, 0.2 mM each dNTP, 0.2 mM each primer, and 2.5 U
of Taq DNA polymerase (Promega). Ten microliters of the
extracted DNA was added to the mixture in all cases, but the
reactions were performed directly on intact cells without
DNA purification steps, cells being taken directly from the
solid media with a sterile plastic tip and resuspended in the
PCR mixture. PCR was performed in a thermocycler (Pro-
gene, Techne) as follows. The first cycle included 3 min of


c 2006 Federation of European Microbiological Societies
Published by Blackwell Publishing Ltd. All rights reserved
A. Galán et al.
denaturation at 94 1C. This first step was followed by 30
cycles of 40 s of denaturation at 94 1C, 30 s of annealing at
60 1C, and 2 min of primer extension at 72 1C. The last step
included one cycle of 40 s of denaturation and a final
extension step of 12 min at 72 1C. A 10-mL aliquot of the
amplified product was analyzed on 0.8% agarose gel and
stained with a 50 mg mL1 ethidium bromide solution.
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Appropriate positive and negative controls were included,
and methods to prevent PCR contamination were used
(Kwok & Higuchi, 1989).
Southern blot analysis
Twenty micrograms of genomic DNA were digested with
BamHI, and fragments were separated by electrophoresis in
agarose gel (0.8%). The gel was stained with ethidium
bromide, and then denatured and neutralized (Sambrook
et al., 1989). The DNA was transferred to a Hybond nylon
membrane (Roche) and hybridized with the KER1 probe
after nonradioactive labeling using the Dig DNA Labeling
and Detection kit (Roche). Hybridization was carried out
under high-stringency conditions following the manufac-
turer’s recommendations.
Results and discussion
In this study, we describe a rapid, simple and reliable PCR
system for the definitive identification of C. albicans. In
general, protocols for extraction of DNA of fungal cells are
either very time-consuming or give poor yields compared to
methods of DNA extraction from human cells or viruses.
Accordingly, several DNA extraction kits have been devel-
oped (Löffler et al., 1997) in order to optimize and simplify
the DNA extraction step, prior to PCR applications. How-
ever, low cost per assay is necessary when clinical PCR
applications are required. In this work, a new PCR assay is
described that allows the practical application of this tech-
nique in daily clinical practice by optimization of the two
steps involved, fungal DNA extraction from clinical samples
and the use of a C. albicans-specific probe.
Six different (in-house) DNA extraction methods that are
relatively easy and quick have been tested. Two methods, D
and F, gave positive amplification rates when two different
C. albicans reference strains were used, as shown in Table 2.
Protocol F, based on alkali, detergent and heat treatment for
fungal DNA release, was the method of choice, as it is
simple, quick and cheap, factors that are fundamental in
clinical routine. The advantages of this method are that it is
simpler and quicker than those described previously for the
recovery of Candida DNA from clinical isolates; it does not
require the use of toxic chemicals (phenol/chloroform,
guanidinium thiocyanate) or expensive enzymes (proteinase
K, Zymolyase); and it avoids the need to perform a time-
consuming mechanical disruption step. The reproducibility
FEMS Yeast Res 6 (2006) 1094–1100
Rapid PCR-based test for identifying Candida albicans 1097

Table 2. Comparison of different DNA extraction methods for PCR

i
nd

C. araps osis
is

R. ctis ans
ilos
amplification, using two different reference strains of Candida albicans

Y. mbe e
kef mo

K. ofor is

po sia

ca

ns
a
il

m
a

C. abrat
C. raps

c ina
C.n pica
C. uillier
C. abrat

C. polyti
C. rusei

S. erevi

i ca
Amplicons Amplicons

C. yr

tro r
y

r
ma
kef

alb
pa
DNA with with

gl
la
g

p
l

li
C.g

C.

S.
extraction SC5314 CECT1675 St
Method Protocol description Duration strain strain

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A Lysis buffer1Proteinase 90 min 1/3 0/3
K1Heat
B Lysis buffer1Zymolyase 60 min 0/3 1/3
C Lysis 60 min 1/3 2/3
buffer1Zymolyase1Heat
D Glass beads1Heat 40 min 6/6 6/6
E Intact cells – 6/6 3/6 670 bp
F Alkali1Heat 30 min 6/6 6/6

1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16

of this procedure was found to be high, and its sensitivity Fig. 1. PCR-generated DNA products deduced from the Candida albi-
was in the same range as that of the other extraction cans KER1 gene. DNA samples from different fungal strains were
methods reported in the literature with which it was obtained by method F (see Material and methods) and amplified by PCR
using primers SCF1 and SCR1. Only the DNA sample from C. albicans
compared. Thus, a concentration between 1000 and
gave positive amplification with the selected pair of primers.
100 cells mL1 in the assay mixture was required to obtain
positive amplicons.
To determine the specificity of these primers (SC1F and (a)
St C. albicans clinical isolates
SC1R), Candida and non-Candida species (Table 1), includ-
ing both clinical isolates and reference strains, were tested.
With the same DNA extraction and PCR protocol, all fungal
strains listed in Table 1 failed to give any PCR amplification 670 bp
product with the SC1F and SC1R primers, except C. albicans
(Fig. 1). Although C. albicans is known to be the most
75

virulent of the medically important Candida species (Cal- (b)

16
14
CT

derone, 2002), some of the emerging non-C. albicans

C1
53

N7

C. albicans clinical isolates

CE
SC

CA
C1

species, such as C. glabrata and C. krusei (Nguyen et al.,

1996; Pfaller et al., 1998, 1999), show reduced susceptibility
to commonly used antifungal agents, thus making necessary
a discriminatory method for identification to the species
level so that optimum antifungal therapy can be determined
(Sullivan et al., 1996).
The basic aim of this study was to evaluate the diagnostic
potential of the KER1-specific primers in routine clinical
work. To further check this possibility, both primers (SC1F
and SC1R) were used to amplify DNA samples from
C. albicans isolates. One hundred and fifty clinical strains
previously identified as C. albicans by standard microbiolo-
gical methods, such as the sugar assimilation test (API 20C
AUX Bio Merieux), germ tube formation and agglutination
tests (see Materials and methods), were tested. The expected
670-bp DNA fragment was amplified from 142 clinical
isolates. The amplified products of some positive C. albicans
samples are shown in Fig. 2. Identification of C. albicans, by
either PCR (Fig. 2a) or Southern hybridization (Fig. 2b),
was in agreement with the traditional identification in all
cases except for eight isolates that were subcultured on
CHROMagar plates for phenotypic diagnosis. CHROMagar
Candida allows differentiation among yeasts on the basis of
3.8 kb
Fig. 2. (a) DNA amplifications from clinical isolates by PCR. DNA from
Candida albicans clinical strains isolated from systemic and mucocuta-
neous infections was used as DNA template for PCR with the SC1F and
SC1F primers. (b) Southern blot analysis of digested (with BamHI)
genomic DNA from C. albicans SC5314 (lane 1), CECT1675 (lane 2),
C1N7 (lane 3) and CAC1 (lane 4) and from several C. albicans clinical
isolates (lanes 5–9). The DNA fragment derived from the KER1 sequence,
amplified by PCR using primers SCF1 and SCR1, was used as a probe.
strongly contrasted colors produced by reactions of species-
specific enzymes with a proprietary chromogenic substrate
(Odds & Bernaerts, 1994). The eight colonies inoculated in
this medium showed a pattern completely different from that
assigned for C. albicans (Fig. 3). These eight clinical isolates
(kindly provided by Dr J. Peman from the Servicio de
Microbiologı́a, Hospital Universitario La Fe, Valencia, Spain)
were further characterized as C. glabrata on the basis of their
pattern of sugar assimilation (auxonogram) with the API 20C

FEMS Yeast Res 6 (2006) 1094–1100

c2006 Federation of European Microbiological Societies
Published by Blackwell Publishing Ltd. All rights reserved
1098
C. albicans
SC5314
Fig. 3. Identification of non-Candida albicans clinical isolates by inocu-
lation on CHROMagar plates.
AUX system. These results support the discriminatory power
of the method described here and its potential diagnostic
value; thus, the specificity of the PCR assay is likely to be
100%. The discriminatory potential of KER1 could also be
considered for the detection of C. albicans in some cases of
multiple infections of Candida and non-Candida spp.
The emergence of C. dubliniensis as a potential pathogen,
especially in immunodepressed (mainly HIV-infected) pa-
tients (Coleman et al., 1993; McCullough et al., 1995;
Coleman et al., 1997), and the fact that C. albicans and C.
dubliniensis are phenotypically very similar, highlights the
need for rapid methods of discriminating between these two
species, which are currently identified by time-consuming
standard microbiological and biochemical procedures.
Although C. dubliniensis is not currently listed as one of the
yeasts identified in the API 20C database, differences be-
tween C. albicans and C. dubliniensis in the assimilation of
xylose and a-methyl-D-glucoside (Salkin et al., 1998; Gales
et al., 1999), and other physical (growth temperature) or
chemical (b-glucosidase activity) tests, may be useful for the
phenotypic differentiation of these species (Pinjon et al.,
1998; Sullivan & Coleman, 1998). However, C. dubliniensis
isolates form a genetically homogeneous group of organisms
that possess a unique genomic organization (Sullivan et al.,
1995), allowing molecular differentiation from C. albicans.
To investigate whether KER1 sequence differences might
be useful in terms of differentiation between these two
species, we performed PCR with the primers derived from
the KER1 sequence of C. albicans. DNA was extracted from
several C. dubliniensis isolates (also provided by Dr J.
Peman) as reported elsewhere, and each experiment in-
cluded positive and negative controls. As the assignment of
isolates to C. dubliniensis is based on lack of amplification of
the KER1 gene, proper adjustment of the controls was
necessary. Template DNA from the C. albicans isolates led
to the specific amplification of a fragment with the predicted


c 2006 Federation of European Microbiological Societies
Published by Blackwell Publishing Ltd. All rights reserved
A. Galán et al.
(a)
75
16
14
CT
C1
53
N7
C. dubliniensis
SC
CA
CE
C1
670 bp
Downloaded from https://academic.oup.com/femsyr/article/6/7/1094/491946 by Semenov Institite for Chemical Physics user on 24 June 2023
(b)
75
16
14
CT
C1
53
N7
C. dubliniensis
SC
CE
CA
C1
3.8 kb
Fig. 4. Capacity of primers from the KER1 gene to discriminate between
Candida dubliniensis and Candida albicans. Genomic DNA from several
clinical isolates of C. dubliniensis (lanes 1–5) and C. albicans (lanes 6–9)
were analyzed by PCR with primers SCF1F and SC1R (a) and Southern
blot with the SC1 gene probe (b). Candida albicans SC5314 and
CECT1675 (lanes 6 and 7) and C1N7 and CAC1 (lanes 8 and 9) were
used as positive and negative controls, respectively.
size of approximately 670 bp. When the same conditions
were used with the five C. dubliniensis isolates, no amplimers
were detected (Fig. 4a). Accordingly, with PCR, no bands
were recognized by Southern blot in C. dubliniensis samples,
whereas the predicted size was obtained for C. albicans
samples (Fig. 4b), thus confirming the potential diagnostic
value of the proposed method.
Deduced primers from the coding region of the C.
albicans-specific KER1 gene, SC1F and SC1R, amplified the
expected 670-bp fragment, using as template the DNA
obtained by following protocol F for both C. albicans
reference strains assayed (CECT1675 and SC5314), and
failed to amplify the C. albicans null mutant strains for the
KER1 gene (Fig. 4a).
Given the rapidly fatal course of acute, disseminated
candidiasis (Beck-Sagué & Jarvis, 1993; Reiss & Morrison,
1993; Jarvis, 1995), a rapid and more accurate method for
diagnosing candidemia could result in significant savings
and benefits to patients and hospitals in lives lost, inap-
propriate or unnecessary drug administration, and hospita-
lization time. In this context, the PCR-based identification
method described here, using primers from the KER1 gene
(Galán et al., 2004) in combination with a simple and fast
DNA extraction protocol, could be useful for the unequi-
vocal identification of C. albicans. Also, the identification of
C. albicans using these specific primers would avoid cross-
reactivity with bacteria, other fungi or other yeast species,
and possible interference when using blood cultures con-
taining bacteria, as there is no homology of the KER1 gene
sequence with any other known sequence.
FEMS Yeast Res 6 (2006) 1094–1100
Rapid PCR-based test for identifying Candida albicans
Acknowledgements
This work was supported by grants BFU2005-02572, Minis-
terio de Educación y Ciencia, Spain, and ACOMP06/103,
Generalitat Valenciana, Valencia, Spain (to J.P.M.). A.G. and
V.V. were the recipients of a predoctoral grant from Minis-
terio de Educación y Ciencia, Spain.
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