Identification of dermatophytes by an oligonucleotide array.
Identifieur interne : 000539 ( Ncbi/Checkpoint ); précédent : 000538; suivant : 000540Identification of dermatophytes by an oligonucleotide array.
Auteurs : Hsin Chieh Li [République populaire de Chine] ; Jean-Philippe Bouchara ; Mark Ming-Long Hsu ; Richard Barton ; Tsung Chain ChangSource :
- Journal of clinical microbiology [ 0095-1137 ] ; 2007.
Descripteurs français
- KwdFr :
- MESH :
- génétique : Arthrodermataceae.
- isolement et purification : Arthrodermataceae.
- Espaceur de l'ADN ribosomique, Humains, Sensibilité et spécificité, Sondes oligonucléotidiques, Séquençage par oligonucléotides en batterie.
English descriptors
- KwdEn :
- MESH :
- chemical , chemistry : DNA, Ribosomal Spacer.
- genetics : Arthrodermataceae.
- isolation & purification : Arthrodermataceae.
- methods : Oligonucleotide Array Sequence Analysis.
- Humans, Oligonucleotide Probes, Sensitivity and Specificity.
Abstract
Species of dermatophytes are classified into three anamorphic (asexual) genera, Epidermophyton, Microsporum, and Trichophyton. Conventional methods used to identify dermatophytes are often lengthy and may be inconclusive because of atypical microscopic or colony morphology. Based on the internal transcribed spacer 1 (ITS-1) and ITS-2 sequences of the rRNA genes, an oligonucleotide array was developed to identify 17 dermatophyte species. The method consisted of PCR amplification of the ITS regions using universal primers, followed by hybridization of the digoxigenin-labeled PCR products to an array of oligonucleotides (17- to 30-mers) immobilized on a nylon membrane. Of 198 dermatophyte strains and 90 nontarget strains tested, the sensitivity and specificity of the array were 99.5% and 97.8%, respectively. The only strain not identified (Microsporum audouinii LMA 597) was found to have a nucleotide insertion at the ITS-2 region where the probe was designed. Two nontarget strains, Microsporum equinum LMA 40396666 and Trichophyton gourvilii var. intermedium CBS 170.65, were misidentified as Microsporum canis and Trichophyton soudanense, respectively. Sequence analysis of the ITS regions revealed that the two misidentified strains displayed high sequence homology with the probes designed for M. canis and T. soudanense, respectively. The present method can be used as a reliable alternative to conventional identification methods and can be completed with isolated colonies within 24 h.
DOI: 10.1128/JCM.00829-07
PubMed: 17687010
Affiliations:
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sortKey="Hsu, Mark Ming Long" sort="Hsu, Mark Ming Long" uniqKey="Hsu M" first="Mark Ming-Long" last="Hsu">Mark Ming-Long Hsu</name></author><author><name sortKey="Barton, Richard" sort="Barton, Richard" uniqKey="Barton R" first="Richard" last="Barton">Richard Barton</name></author><author><name sortKey="Chang, Tsung Chain" sort="Chang, Tsung Chain" uniqKey="Chang T" first="Tsung Chain" last="Chang">Tsung Chain Chang</name></author></titleStmt><publicationStmt><idno type="wicri:source">PubMed</idno><date when="2007">2007</date><idno type="RBID">pubmed:17687010</idno><idno type="pmid">17687010</idno><idno type="doi">10.1128/JCM.00829-07</idno><idno type="wicri:Area/PubMed/Corpus">002167</idno><idno type="wicri:explorRef" wicri:stream="PubMed" wicri:step="Corpus" wicri:corpus="PubMed">002167</idno><idno type="wicri:Area/PubMed/Curation">002167</idno><idno type="wicri:explorRef" wicri:stream="PubMed" wicri:step="Curation">002167</idno><idno 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Tainan 701, Taiwan</wicri:regionArea><wicri:noRegion>Taiwan</wicri:noRegion></affiliation></author><author><name sortKey="Bouchara, Jean Philippe" sort="Bouchara, Jean Philippe" uniqKey="Bouchara J" first="Jean-Philippe" last="Bouchara">Jean-Philippe Bouchara</name></author><author><name sortKey="Hsu, Mark Ming Long" sort="Hsu, Mark Ming Long" uniqKey="Hsu M" first="Mark Ming-Long" last="Hsu">Mark Ming-Long Hsu</name></author><author><name sortKey="Barton, Richard" sort="Barton, Richard" uniqKey="Barton R" first="Richard" last="Barton">Richard Barton</name></author><author><name sortKey="Chang, Tsung Chain" sort="Chang, Tsung Chain" uniqKey="Chang T" first="Tsung Chain" last="Chang">Tsung Chain Chang</name></author></analytic><series><title level="j">Journal of clinical microbiology</title><idno type="ISSN">0095-1137</idno><imprint><date when="2007" type="published">2007</date></imprint></series></biblStruct></sourceDesc></fileDesc><profileDesc><textClass><keywords scheme="KwdEn" xml:lang="en"><term>Arthrodermataceae (genetics)</term><term>Arthrodermataceae (isolation & purification)</term><term>DNA, Ribosomal Spacer (chemistry)</term><term>Humans</term><term>Oligonucleotide Array Sequence Analysis (methods)</term><term>Oligonucleotide Probes</term><term>Sensitivity and Specificity</term></keywords><keywords scheme="KwdFr" xml:lang="fr"><term>Arthrodermataceae (génétique)</term><term>Arthrodermataceae (isolement et purification)</term><term>Espaceur de l'ADN ribosomique ()</term><term>Humains</term><term>Sensibilité et spécificité</term><term>Sondes oligonucléotidiques</term><term>Séquençage par oligonucléotides en batterie ()</term></keywords><keywords scheme="MESH" type="chemical" qualifier="chemistry" xml:lang="en"><term>DNA, Ribosomal Spacer</term></keywords><keywords scheme="MESH" qualifier="genetics" xml:lang="en"><term>Arthrodermataceae</term></keywords><keywords scheme="MESH" qualifier="génétique" xml:lang="fr"><term>Arthrodermataceae</term></keywords><keywords scheme="MESH" qualifier="isolation & purification" xml:lang="en"><term>Arthrodermataceae</term></keywords><keywords scheme="MESH" qualifier="isolement et purification" xml:lang="fr"><term>Arthrodermataceae</term></keywords><keywords scheme="MESH" qualifier="methods" xml:lang="en"><term>Oligonucleotide Array Sequence Analysis</term></keywords><keywords scheme="MESH" xml:lang="en"><term>Humans</term><term>Oligonucleotide Probes</term><term>Sensitivity and Specificity</term></keywords><keywords scheme="MESH" xml:lang="fr"><term>Espaceur de l'ADN ribosomique</term><term>Humains</term><term>Sensibilité et spécificité</term><term>Sondes oligonucléotidiques</term><term>Séquençage par oligonucléotides en batterie</term></keywords></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en">Species of dermatophytes are classified into three anamorphic (asexual) genera, Epidermophyton, Microsporum, and Trichophyton. Conventional methods used to identify dermatophytes are often lengthy and may be inconclusive because of atypical microscopic or colony morphology. Based on the internal transcribed spacer 1 (ITS-1) and ITS-2 sequences of the rRNA genes, an oligonucleotide array was developed to identify 17 dermatophyte species. The method consisted of PCR amplification of the ITS regions using universal primers, followed by hybridization of the digoxigenin-labeled PCR products to an array of oligonucleotides (17- to 30-mers) immobilized on a nylon membrane. Of 198 dermatophyte strains and 90 nontarget strains tested, the sensitivity and specificity of the array were 99.5% and 97.8%, respectively. The only strain not identified (Microsporum audouinii LMA 597) was found to have a nucleotide insertion at the ITS-2 region where the probe was designed. Two nontarget strains, Microsporum equinum LMA 40396666 and Trichophyton gourvilii var. intermedium CBS 170.65, were misidentified as Microsporum canis and Trichophyton soudanense, respectively. Sequence analysis of the ITS regions revealed that the two misidentified strains displayed high sequence homology with the probes designed for M. canis and T. soudanense, respectively. The present method can be used as a reliable alternative to conventional identification methods and can be completed with isolated colonies within 24 h.</div></front></TEI><affiliations><list><country><li>République populaire de Chine</li></country></list><tree><noCountry><name sortKey="Barton, Richard" sort="Barton, Richard" uniqKey="Barton R" first="Richard" last="Barton">Richard Barton</name><name sortKey="Bouchara, Jean Philippe" sort="Bouchara, Jean Philippe" uniqKey="Bouchara J" first="Jean-Philippe" last="Bouchara">Jean-Philippe Bouchara</name><name sortKey="Chang, Tsung Chain" sort="Chang, Tsung Chain" uniqKey="Chang T" first="Tsung Chain" last="Chang">Tsung Chain Chang</name><name sortKey="Hsu, Mark Ming Long" sort="Hsu, Mark Ming Long" uniqKey="Hsu M" first="Mark Ming-Long" last="Hsu">Mark Ming-Long Hsu</name></noCountry><country name="République populaire de Chine"><noRegion><name sortKey="Li, Hsin Chieh" sort="Li, Hsin Chieh" uniqKey="Li H" first="Hsin Chieh" last="Li">Hsin Chieh Li</name></noRegion></country></tree></affiliations></record>Pour manipuler ce document sous Unix (Dilib)
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