Previous Article | Next Article 
Journal of Clinical Microbiology, April 1999, p. 920-924, Vol. 37, No. 4
0095-1137/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.
Phylogenetic Classification and Species Identification of
Dermatophyte Strains Based on DNA Sequences of Nuclear Ribosomal
Internal Transcribed Spacer 1 Regions
Koichi
Makimura,1,*
Yoshiko
Tamura,1
Takashi
Mochizuki,2
Atsuhiko
Hasegawa,3
Yoshito
Tajiri,1
Ryo
Hanazawa,1
Katsuhisa
Uchida,1
Hiuga
Saito,1 and
Hideyo
Yamaguchi1
Teikyo University Institute of Medical
Mycology, Tokyo,1 Department of
Dermatology, Kanazawa Medical University,
Ishikawa,2 and Department of Veterinary
Medicine, Nihon University, Kanagawa,3 Japan
Received 4 September 1998/Returned for modification 5 November
1998/Accepted 24 December 1998
 |
ABSTRACT |
The mutual phylogenetic relationships of dermatophytes of the
genera Trichophyton, Microsporum, and
Epidermophyton were demonstrated by using internal
transcribed spacer 1 (ITS1) region ribosomal DNA sequences.
Trichophyton spp. and Microsporum spp. form a
cluster in the phylogenetic tree with Epidermophyton
floccosum as an outgroup, and within this cluster, all
Trichophyton spp. except Trichophyton terrestre
form a nested cluster (100% bootstrap support). Members of
dermatophytes in the cluster of Trichophyton spp. were
classified into three groups with ITS1 homologies, with each of them
being a monophyletic cluster (100% bootstrap support). The
Arthroderma vanbreuseghemii-Arthroderma simii group
consists of A. vanbreuseghemii, A. simii,
Trichophyton mentagrophytes isolates from humans, T. mentagrophytes var. quinckeanum, Trichophyton
tonsurans, and Trichophyton schoenleinii. Arthroderma
benhamiae, T. mentagrophytes var.
erinacei, and Trichophyton verrucosum are
members of the Arthroderma benhamiae group.
Trichophyton rubrum and Trichophyton violaceum
form the T. rubrum group. This suggests that these
"species" of dermatophytes have been overclassified. The ITS1
sequences of 11 clinical isolates were also determined to identify the
species, and all strains were successfully identified by comparison of
their base sequences with those in the ITS1 DNA sequence database.
| |
INTRODUCTION |
Dermatophytes (dermatomycetes) have
the capacity to invade keratinized tissues of humans and other animals
to produce an infection, dermatophytosis (dermatomycosis)
(28). The phylogeny of dermatophytes, however, remains
unclear because their members are phylogenetically and taxonomically
very closely related, their phenotypic features are sometimes poor, and
many isolates from medical and veterinary samples have lost their
sexual activity (25). From a clinical point of view, for
definition of species or for performance of an epidemiological study,
it is important to have a reliable method for the identification of
dermatophyte species. Molecular biological studies of the phylogeny of
the fungi have been performed, primarily by using the G+C content of
chromosomal DNA (4), total DNA homology (5),
restriction fragment length polymorphism (RFLP) analysis of
mitochondrial DNA (mtDNA) (6, 14, 15, 21, 23), random
amplification of polymorphic DNA (10, 13, 17, 22), and
determination of the base sequence of 18S (11) or 28S
(16) rRNA or ribosomal DNA (rDNA). For dermatophytes,
however, the phylogenetic relationships of species or species-specific sequences cannot be fully defined by these methods.
Specific DNA sequences of internal transcribed spacer (ITS) 1 (ITS1) of
rDNA in the dermatophytes were therefore determined and were analyzed
phylogenetically. ITS1 is located between the 18S and the 5.8S rDNAs.
As reported previously, the variable ITS regions have been proven to be
useful in resolving relationships between close taxonomic relatives
(2, 3, 18), and in the field of medical mycology, several
phylogenetic studies in which the ITS1 region and the primer system
designed by Makimura et al. (20) or White et al.
(29) were used were reported on recently (1, 20, 26,
27). We have reported that it is feasible to successfully
differentiate between members of the Trichophyton mentagrophytes complex, the major dermatophytes, which are hard to
identify by their morphological features, by demonstrating their
phylogenetic relationship by comparing the base pair sequences of the
ITS1 regions (20). In the present study we determined the
phylogeny of the group of dermatophytes, including the genera Trichophyton, Microsporum, and
Epidermophyton, and identified the species using the base
pair sequences of ITS1.
| |
MATERIALS AND METHODS |
Fungal strains.
The 12 standard strains and 11 clinical
isolates of dermatophytes used in this study are described in Tables
1 and 2.
The clinical strains were isolated in Japan and were identified by their morphological features, but the species of two of them could not
be specified because they did not have typical microscopic structures.
Preparation of DNA from fungal cells.
All fungal strains
were grown on Sabouraud dextrose agar (peptone, 1% [wt/vol];
glucose, 1% [wt/vol]; agar, 1.5% [wt/vol]) at 27°C for 5 days.
Rapid preparation of DNA from strains was performed by the method
described by the authors (20). A small amount of mycelium
grown on Sabouraud dextrose agar was placed in lysis buffer (200 mM
Tris-HCl [pH 8.0], 0.5% [wt/vol] sodium dodecyl sulfate, 250 mM
NaCl, 25 mM EDTA) and crushed with a conical grinder. It was then
incubated at 100°C for 15 min and mixed with 150 µl of 3.0 M sodium
acetate, kept at 
20°C for 10 min, and then centrifuged at
10,000 × g for 5 min. The supernatant was extracted
once with phenol-chloroform-isoamyl alcohol (25:24:1 [vol/vol]) and
was subsequently extracted once with chloroform. The DNA was
precipitated with an equal volume of isopropanol at 20°C for 10 min, washed with 0.5 ml of 99% ethanol, dried, and suspended in 50 µl of ultrapure water (Milli-Q Synthesis A10; Millipore). One
microliter of solution was used as the template for PCR. The total time
required to prepare the DNA was 80 min.
Oligonucleotides.
The oligonucleotide primers, designed by
the authors (20), (18SF1, 5'-AGGTTTCCGTAGGTGAACCT-3';
58SR1, 5'-TTCGCTGCGTTCTTCATCGA-3') were made by
Amersham Pharmacia Biotech Co., Ltd. (Tokyo, Japan).
PCR.
Each PCR mixture contained 10 µl of 10× reaction
buffer (Pharmacia), 100 µM (each) dATP, dCTP, dGTP, and dTTP
(Pharmacia), 2.5 U of Taq polymerase (Pharmacia), 30 pmol of
each primer, and DNA template solution. Ultrapure water was added to
increase the volume to 100 µl. Each reaction mixture was heated to
94°C for 5 min, and PCR was performed under the following conditions:
94°C for 1 min, 60°C for 15 s, and 72°C for 15 s for 25 cycles. The thermal cycles were terminated by polymerization at 72°C
for 10 min. The products were detected as a single band of 0.3 kbp by agarose gel electrophoresis and UV irradiation.
ITS1 DNA sequencing and phylogenetic analysis.
Both strands
of the PCR products were directly sequenced with a DNA Sequencing Kit
(Perkin-Elmer) with primers 18SF1 and 58SR1 and an automatic sequencer
(Genetic Analyzer 310; Perkin-Elmer), according to the manufacturer's instructions.
The ITS1 sequences of the standard strains used in this study and of
members of the T. mentagrophytes complex (Arthroderma vanbreuseghemii, DDBJ/EMBL/GenBank accession no. AB011488; Arthroderma benhamiae [Americano-European race], accession
no. AB011457; Arthroderma benhamiae [African race],
accession no. AB011454; Arthroderma simii, accession no.
AB011461; T. mentagrophytes isolates from humans, accession
no. AB011463; T. mentagrophytes var.
erinacei, accession no. AB011455; Trichophyton rubrum, accession no. AB011453), as reported by the
authors (20), were aligned by using the Clustal W computer
program (12) and GENETYX-MAC 10.1 software (Software
Development Co., Ltd., Tokyo, Japan). Phylogenetic trees were then
constructed by the DNA maximum-likelihood (ML) method in the PHYLIP
program (Phylogeny Inference Package), version 3.5c (8), and
the neighbor-joining (NJ) (24) method in the NJPLOT program
(9). Bootstrap (7) analysis with the Clustal W
program was performed by taking 1,000 random samples from the multiple
alignment. This provided a measure of how well supported parts of the
tree are, given the data set and the method used to construct the tree.
The tree was rooted with Epidermophyton floccosum as an
outgroup, because it was shown that this species is phylogenetically
distant from other dermatophytes by the RFLP analysis of mtDNA
(15). The evolutionary distance between organisms is
indicated by the horizontal branch length, which reflects the number of
nucleotide substitutions per site along that branch from the node to
the endpoint. In the NJ tree, the percentage of bootstrap samplings
that support the interior branches is noted.
Nucleotide sequence accession numbers.
The nucleotide
sequence data reported in this paper will appear in the
DDBJ/EMBL/GenBank nucleotide sequence database with the accession
numbers presented in Table 1.
| |
RESULTS |
Each strain of the dermatophytes tested was shown to have unique
ITS1 base sequences, and the two strains of T. mentagrophytes var. quinckeanum were found to be
identical. Phylogenetic trees were prepared by the NJ (Fig.
1) and ML (Fig.
2) methods and were constructed from data
for 18 species of dermatophytes. The sequences of their ITS1 regions
are presented in Fig. 3; the sizes of
these regions ranged from 175 to 293 bp. In the NJ tree,
Trichophyton spp. and Microsporum spp. form
cluster A, and all Trichophyton spp. except
Trichophyton terrestre form cluster B (100% bootstrap support). The members of the dermatophytes in cluster B were classified into three groups with ITS1 homology (groups a, b, and c) according to
their ITS1 DNA sequences, and each of them is a monophyletic cluster
(100% bootstrap support). ITS1 homology group a (A. vanbreuseghemii-A. simii group) consists of A. vanbreuseghemii, A. simii, T. mentagrophytes isolates from humans, T. mentagrophytes var.
quinckeanum, Trichophyton tonsurans, and
Trichophyton schoenleinii. Both races of A. benhamiae, T. mentagrophytes var.
erinacei, and Trichophyton verrucosum are members
of ITS1 homology group b (A. benhamiae group). T. rubrum and Trichophyton violaceum form a cluster of
ITS1 homology group c (T. rubrum group). The
phylogenetic relationships mentioned above were also supported by
the ML tree.
View larger version (18K):
[in this window]
[in a new window]
|
FIG. 1.
NJ tree of dermatophytes on the basis of their ITS1
sequences. The NJ tree was constructed with data for standard strains
of dermatophytes (see Table 1 and the text). The numbers above the
branches indicate the percentage of bootstrap samplings. Branches
without numbers have frequencies of less than 70%. A, cluster of
Trichophyton spp. and Microsporum spp.; B,
cluster of all Trichophyton spp. except T. terrestre; a, Arthroderma vanbreuseghemii-A. simii
group; b, A. benhamiae group; c, T. rubrum group.
K nuc, thousands of nucleotides.
|
|
View larger version (14K):
[in this window]
[in a new window]
|
FIG. 2.
ML tree of dermatophytes on the basis of their ITS1
sequences. The ML tree was constructed with data for standard strains
of dermatophytes (see Table 1 and the text). A, cluster of
Trichophyton spp. and Microsporum spp.; B,
cluster of all Trichophyton spp. except T. terrestre; a, A. vanbreuseghemii-A. simii group; b,
A. benhamiae group; c, T. rubrum group.
K nuc, thousands of nucleotides.
|
|
View larger version (68K):
[in this window]
[in a new window]
|
FIG. 3.
Alignment of ITS1 sequences of dermatophytes. The
sequences of 18 species of dermatophytes (see Table 1 and the text)
were aligned by using the Clustal W (12) and GENETYX-MAC
10.1 (Software Development Co., Ltd.) computer programs. Hyphens
designate gaps that were added to permit alignment. T. men.
quinckeanum, T. mentagrophytes var.
quinckeanum; T. men. erinacei, T. mentagrophytes var. erinacei; A. benhamiae
A/E and Af, A. benhamiae Americano-European race and African
race, respectively.
|
|
The ITS1 sequences of 11 clinical isolates were also determined. Each
of the morphologically identified strains of dermatophytes (three
strains of T. rubrum, four strains of T. mentagrophytes, one strain of T. violaceum, and
one strain of Microsporum canis) was shown to have ITS1 base
pair sequences identical to that of the respective standard
strain tested (Table 2). Two morphologically unidentifiable
strains were then subjected to ITS1 sequencing; one was revealed to be
T. rubrum, and the other was shown to be T. violaceum on the basis of the ITS1 DNA sequence database
constructed by the authors.
| |
DISCUSSION |
Using ITS1 rDNA sequences from 12 newly sequenced and 7 previously reported strains of fungi, we have described the phylogeny of members of the dermatophytes. The phylogenetic relationship based on
the ITS1 DNA sequence alignment of meiosporic (perfect) and mitosporic
(imperfect) states of the strains agreed with the proposed taxonomic
connection in their sexual compatibility (25), RFLP analysis
of mtDNA (14, 15, 21, 23), and a phylogenetic study based on
18S rDNA (11) or 28S rDNA (16) sequences. In particular, the last two papers dealt with the base sequences of
the rDNA region, but they omitted A. benhamiae, T. verrucosum, and T. mentagrophytes var.
erinacei, which together formed a unique cluster, cluster
B-b, in the phylogenetic trees that appear in Fig. 1 and 2. Thus,
because of their highly variable ITS1 sequences, the phylogenetic
analysis of the members of the dermatophytes was achieved in more
accurate detail.
We stated earlier that there were three ITS1 homology groups (groups a,
b, and c) in the cluster of Trichophyton spp. The clusters
of ITS1 homology group a (A. vanbreuseghemii-A. simii group), group b (A. benhamiae group), and group c (T. rubrum group) were 100% supported by bootstrap analysis. A. vanbreuseghemii, A. simii, A. benhamiae, and
anamorphic species of T. mentagrophytes constitute the
T. mentagrophytes complex (20, 25) because they
are difficult to distinguish from each other by their morphological features. By using sexual compatibility (25), RFLP analysis of mtDNA (21), or DNA sequence analysis of ITS1
(20), the T. mentagrophytes complex was shown to
be monophyletic, and each of the members was identified. The present
study showed that the species of dermatophytes pathogenic for humans or
animals, T. tonsurans, T. schoenleinii, and
T. verrucosum, are members of the A. vanbreuseghemii-A. simii group (Fig. 1, cluster a) or the A. benhamiae group (Fig. 1, cluster b). This suggests that these medically important dermatophytes have been overclassified. However, since each of the "species" has a unique phenotype, pathogenicity, and host-specific affinity, it is reasonable to retain their
"species" identifications in order to identify the pathogen.
In addition to establishing the significance of the ITS1 region from a
taxonomic standpoint, we also identified these clinically important
species using the ITS1 DNA sequence database. With this system, not
only the morphologically identified strains of T. mentagrophytes, T. rubrum, T. violaceum, and
M. canis but also the two strains of morphologically
unidentifiable strains, T. rubrum and T. violaceum, were successfully identified. This ITS1-based identification system saves time (it takes 2 to 3 days) and is accurate
and applicable even to strains with atypical morphological features.
Further evaluation of the phylogenetic analysis and identification
system, both of which are based on ITS1 rDNA sequences, is under way in
our laboratory with other species and strains. Moreover, a new approach
to the detection and identification of pathogenic fungi from clinical
specimens, i.e., skin, nail, or hair samples, with this database, along
with previously reported molecular diagnostic systems (19),
is also in progress in our laboratory.
| |
ACKNOWLEDGMENTS |
We thank Takashi Sugita, Department of Microbiology, Meiji
College of Pharmacy, for technical advice.
This study was partly supported by a research grant for bioscience from
the Sapporo Bioscience Foundation of Japan and the Proposal-Based
Advanced Industrial Technology R & D Program (grant B-276) of the New
Energy and Industrial Technology Development Organization (NEDO) of Japan.
| |
FOOTNOTES |
*
Corresponding author. Mailing address: Teikyo
University Institute of Medical Mycology, Otsuka, Hachioji, Tokyo
192-0395 Japan. Phone: 81-426-78-3256. Fax: 81-426-74-9190. E-mail: makimura{at}main.teikyo-u.ac.jp.
| |
REFERENCES |
| 1.
|
Attili, D. S.,
G. S. de Hoog, and A. A. Pizzirani-Kleiner.
1998.
rDNA-RFLP and ITS1 sequencing of species of the genus Fonsecaea, agents of chromoblastomycosis.
Med. Mycol.
36:219-225.
[Medline] |
| 2.
|
Berbee, M. L.,
A. Yoshimura,
J. Sugiyama, and J. W. Taylor.
1995.
Is Penicillium monophyletic? An evaluation of phylogeny in the family Trichocomaceae from 18S, 5.8S and ITS ribosomal DNA sequence data.
Mycologia
87:210-222.
|
| 3.
|
Carbone, I., and L. M. Kohn.
1993.
Ribosomal DNA sequence divergence within internal transcribed spacer 1 of the Sclerotiniaceae.
Mycologia
85:415-427.
|
| 4.
|
Davidson, F. D.,
D. W. R. Mackenzie, and R. J. Owen.
1980.
Deoxyribonucleic acid base compositions of dermatophytes.
J. Gen. Microbiol.
118:465-470[Abstract/Free Full Text].
|
| 5.
|
Davidson, F. D., and D. W. R. Mackenzie.
1984.
DNA homology studies in the taxonomy of dermatophytes.
Sabouraudia
2:117-123.
|
| 6.
|
de Bièvre, C.,
C. Dauguet,
V. H. Nguyen, and O. Iburahim-Granet.
1987.
Polymorphism in mitochondria DNA of several Trichophyton rubrum isolates from clinical specimens.
Ann. Inst. Pasteur Microbiol.
138:719-727[Medline].
|
| 7.
|
Felsenstein, J.
1985.
Confidence limits on phylogenies: an approach using the bootstrap.
Evolution
39:783-791.
|
| 8.
|
Felsenstein, J.
1995.
PHYLIP (Phylogeny Inference Package), version 3.5c.
University of Washington, Seattle.
|
| 9.
|
Gouy, M.
1995.
NJPLOT.
University of Lyon 1, Lyon, France.
|
| 10.
|
Graser, Y.,
M. el Fari,
W. Presber,
W. Sterry, and H. J. Tietz.
1998.
Identification of common dermatophytes (Trichophyton, Microsporum, Epidermophyton) using polymerase chain reactions.
Br. J. Dermatol.
138:576-582[Medline].
|
| 11.
|
Harmsen, D.,
A. Schwinn,
M. Weig,
E.-B. Bröcker, and J. Heesemann.
1995.
Phylogeny and dating of some pathogenic keratinophilic fungi using small subunit ribosomal RNA.
J. Med. Vet. Mycol.
33:299-303[Medline].
|
| 12.
|
Higgins, D.
1996.
Clustal W, version 1.6.
The European Bioinformatics Institute, Cambridge, United Kingdom.
|
| 13.
|
Kano, R.,
Y. Nakamura,
T. Watari,
S. Watanabe,
H. Takahashi,
H. Tsujimoto, and A. Hasegawa.
1998.
Identification of clinical isolates of Microsporum canis and M. gypseum by random amplification of polymorphic DNA (RAPD) and Southern hybridization analysis.
Mycoses
41:139-143[Medline].
|
| 14.
|
Kawasaki, M.,
M. Aoki,
H. Ishizaki,
K. Nishio,
T. Mochizuki, and S. Watanabe.
1992.
Phylogenetic relationships of the genera Arthroderma and Nanizzia referred from mitochondrial DNA analysis.
Mycopathologia
118:95-102[Medline].
|
| 15.
|
Kawasaki, M.,
M. Aoki,
H. Ishizaki,
K. Nishimura, and M. Miyaji.
1996.
Phylogeny of Epidermophyton floccosum and other dermatophytes.
Mycopathologia
134:121-128[Medline].
|
| 16.
|
Leclerc, M. C.,
H. Philippe, and E. Guého.
1994.
Phylogeny of dermatophytes and dimorphic fungi based on large subunit ribosomal RNA sequence comparisons.
J. Med. Vet. Mycol.
32:331-341[Medline].
|
| 17.
|
Liu, D.,
S. Coloe,
R. Baird, and J. Pedersen.
1997.
Molecular determination of dermatophyte fungi using the arbitrarily primed polymerase chain reaction.
Br. J. Dermatol.
137:351-355[Medline].
|
| 18.
|
Lobuglio, K.,
J. I. Pitt, and J. W. Taylor.
1993.
Phylogenetic analysis of two ribosomal DNA regions indicates multiple independent losses of a sexual Talaromyces state among asexual Penicillium species in subgenus Biverticillium.
Mycologia
85:592-604.
|
| 19.
|
Makimura, K.,
S. Y. Murayama, and H. Yamaguchi.
1994.
Detection of a wide range of medically important fungi by the polymerase chain reaction.
J. Med. Microbiol.
40:358-364[Abstract/Free Full Text].
|
| 20.
|
Makimura, K.,
T. Mochizuki,
A. Hasagawa,
K. Uchida, and H. Yamaguchi.
1998.
Phylogenetic classification of Trichophyton mentagrophytes complex strains based on DNA sequences of nuclear ribosomal internal transcribed spacer 1 regions.
J. Clin. Microbiol.
36:2629-2633[Abstract/Free Full Text].
|
| 21.
|
Mochizuki, T.,
K. Takada,
S. Watanabe,
W. Kawasaki, and H. Ishizaki.
1990.
Taxonomy of Trichophyton interdigitale (Trichophyton mentagrophytes var. interdigitale) by restriction enzyme analysis of mitochondrial DNA.
J. Med. Vet. Mycol.
28:191-196[Medline].
|
| 22.
|
Mochizuki, T.,
N. Sugie, and M. Uehara.
1997.
Random amplification of polymorphic DNA is useful for the differentiation of several anthropophilic dermatophytes.
Mycoses
40:405-409[Medline].
|
| 23.
|
Nishio, K.,
M. Kawasaki, and H. Ishizaki.
1992.
Phylogeny of the genera Trichophyton using mitochondrial DNA analysis.
Mycopathologia
117:127-132[Medline].
|
| 24.
|
Saito, N., and M. Nei.
1987.
The neighbor-joining method: a new method for reconstructing phylogenetic trees.
Mol. Biol. Evol.
4:406-425[Abstract].
|
| 25.
|
Takashio, M.
1977.
The Trichophyton mentagrophytes complex, p. 271-276.
In
K. Iwata (ed.), Recent advances in medical and veterinary mycology. University of Tokyo Press, Tokyo, Japan.
|
| 26.
|
Uijthof, J. M. J.,
A. van Belkum,
G. S. de Hoog, and G. Haase.
1998.
Exophiala deimatitidis and Sarcinomyces phaeomuriformis: ITS1-sequencing and nutritional physiology.
Med. Mycol.
36:143-151.
[Medline] |
| 27.
|
Wedde, M.,
D. Müller,
K. Tintelnot,
G. S. de Hoog, and U. Stahl.
1998.
PCR-based identification of clinically relevant Pseudallescheria/Scedosporium strains.
Med. Mycol.
36:61-67.
[Medline] |
| 28.
|
Weitzman, I., and R. C. Summerbell.
1995.
The dermatophytes.
Clin. Microbiol. Rev.
8:240-259[Abstract].
|
| 29.
|
White, T. J.,
T. Bruns,
S. Lee, and J. Taylor.
1990.
Amplification and direct sequencing of fungal ribosomal RNA genes for phylogenetics.
In
M. A. Innis, D. H. Gelfand, J. J. Sninsky, and T. J. White (ed.), PCR protocols: a guide to methods and applications. Academic Press, San Diego, Calif.
|
Journal of Clinical Microbiology, April 1999, p. 920-924, Vol. 37, No. 4
0095-1137/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.
This article has been cited by other articles:
-
Li, H. C., Bouchara, J.-P., Hsu, M. M.-L., Barton, R., Su, S., Chang, T. C.
(2008). Identification of dermatophytes by sequence analysis of the rRNA gene internal transcribed spacer regions. J Med Microbiol
57: 592-600
[Abstract]
[Full Text]
-
Kong, F., Tong, Z., Chen, X., Sorrell, T., Wang, B., Wu, Q., Ellis, D., Chen, S.
(2008). Rapid Identification and Differentiation of Trichophyton Species, Based on Sequence Polymorphisms of the Ribosomal Internal Transcribed Spacer Regions, by Rolling-Circle Amplification. J. Clin. Microbiol.
46: 1192-1199
[Abstract]
[Full Text]
-
Frealle, E., Rodrigue, M., Gantois, N., Aliouat, C.-M., Delaporte, E., Camus, D., Dei-Cas, E., Kauffmann-Lacroix, C., Guillot, J., Delhaes, L.
(2007). Phylogenetic analysis of Trichophyton mentagrophytes human and animal isolates based on MnSOD and ITS sequence comparison. Microbiology
153: 3466-3477
[Abstract]
[Full Text]
-
Cascio, G. L., Dalle Carbonare, L., Maccacaro, L., Caliari, F., Ligozzi, M., Cascio, V. L., Fontana, R.
(2007). First Case of Bloodstream Infection Due to Candida magnoliae in a Chinese Oncological Patient. J. Clin. Microbiol.
45: 3470-3473
[Abstract]
[Full Text]
-
Li, H. C., Bouchara, J.-P., Hsu, M. M.-L., Barton, R., Chang, T. C.
(2007). Identification of Dermatophytes by an Oligonucleotide Array. J. Clin. Microbiol.
45: 3160-3166
[Abstract]
[Full Text]
-
Binstock, J. M.
(2007). Molecular Biology Techniques for Identifying Dermatophytes and Their Possible Use in Diagnosing Onychomycosis in Human Toenail: A Review. J. Am. Podiatr. Med. Assoc.
97: 134-144
[Abstract]
[Full Text]
-
Gutzmer, R., Mommert, S., Kuttler, U., Werfel, T., Kapp, A.
(2004). Rapid identification and differentiation of fungal DNA in dermatological specimens by LightCycler PCR. J Med Microbiol
53: 1207-1214
[Abstract]
[Full Text]
-
Gaedigk, A., Gaedigk, R., Abdel-Rahman, S. M.
(2003). Genetic Heterogeneity in the rRNA Gene Locus of Trichophyton tonsurans. J. Clin. Microbiol.
41: 5478-5487
[Abstract]
[Full Text]
-
Mochizuki, T., Ishizaki, H., Barton, R. C., Moore, M. K., Jackson, C. J., Kelly, S. L., Evans, E. G. V.
(2003). Restriction Fragment Length Polymorphism Analysis of Ribosomal DNA Intergenic Regions Is Useful for Differentiating Strains of Trichophyton mentagrophytes. J. Clin. Microbiol.
41: 4583-4588
[Abstract]
[Full Text]
-
LIU, D., PEARCE, L., LILLEY, G., COLOE, S., BAIRD, R., PEDERSEN, J.
(2002). PCR identification of dermatophyte fungi Trichophyton rubrum, T. soudanense and T. gourvilii. J Med Microbiol
51: 117-122
[Abstract]
[Full Text]
-
Fujita, S.-I., Senda, Y., Nakaguchi, S., Hashimoto, T.
(2001). Multiplex PCR Using Internal Transcribed Spacer 1 and 2 Regions for Rapid Detection and Identification of Yeast Strains. J. Clin. Microbiol.
39: 3617-3622
[Abstract]
[Full Text]
-
Okeke, C. N., Tsuboi, R., Kawai, M., Hiruma, M., Ogawa, H.
(2001). Isolation of an Intron-Containing Partial Sequence of the Gene Encoding Dermatophyte Actin (ACT) and Detection of a Fragment of the Transcript by Reverse Transcription-Nested PCR as a Means of Assessing the Viability of Dermatophytes in Skin Scales. J. Clin. Microbiol.
39: 101-106
[Abstract]
[Full Text]
-
Gupta, A. K., Kohli, Y., Summerbell, R. C.
(2000). Molecular Differentiation of Seven Malassezia Species. J. Clin. Microbiol.
38: 1869-1875
[Abstract]
[Full Text]
-
Summerbell, R. C., Haugland, R. A., Li, A., Gupta, A. K.
(1999). rRNA Gene Internal Transcribed Spacer 1 and 2 Sequences of Asexual, Anthropophilic Dermatophytes Related to Trichophyton rubrum. J. Clin. Microbiol.
37: 4005-4011
[Abstract]
[Full Text]
Top
Abstract
Introduction
