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Article: Intra-genomic internal transcribed spacer region sequence heterogeneity and molecular diagnosis in clinical microbiology

TitleIntra-genomic internal transcribed spacer region sequence heterogeneity and molecular diagnosis in clinical microbiology
Authors
Issue Date2015
PublisherMolecular Diversity Preservation International. The Journal's web site is located at http://www.mdpi.org/ijms
Citation
International Journal of Molecular Sciences, 2015, v. 16 n. 10, p. 25067-25079 How to Cite?
AbstractInternal transcribed spacer region (ITS) sequencing is the most extensively used technology for accurate molecular identification of fungal pathogens in clinical microbiology laboratories. Intra-genomic ITS sequence heterogeneity, which makes fungal identification based on direct sequencing of PCR products difficult, has rarely been reported in pathogenic fungi. During the process of performing ITS sequencing on 71 yeast strains isolated from various clinical specimens, direct sequencing of the PCR products showed ambiguous sequences in six of them. After cloning the PCR products into plasmids for sequencing, interpretable sequencing electropherograms could be obtained. For each of the six isolates, 10–49 clones were selected for sequencing and two to seven intra-genomic ITS copies were detected. The identities of these six isolates were confirmed to be Candida glabrata (n = 2), Pichia (Candida) norvegensis (n = 2), Candida tropicalis (n = 1) and Saccharomyces cerevisiae (n = 1). Multiple sequence alignment revealed that one to four intra-genomic ITS polymorphic sites were present in the six isolates, and all these polymorphic sites were located in the ITS1 and/or ITS2 regions. We report and describe the first evidence of intra-genomic ITS sequence heterogeneity in four different pathogenic yeasts, which occurred exclusively in the ITS1 and ITS2 spacer regions for the six isolates in this study.
Persistent Identifierhttp://hdl.handle.net/10722/223223
ISSN
2015 Impact Factor: 3.257
2015 SCImago Journal Rankings: 1.160
PubMed Central ID

 

DC FieldValueLanguage
dc.contributor.authorZhao, Y-
dc.contributor.authorTsang, CC-
dc.contributor.authorXiao, M-
dc.contributor.authorCheng, J-
dc.contributor.authorXu, Y-
dc.contributor.authorLau, SKP-
dc.contributor.authorWoo, PCY-
dc.date.accessioned2016-02-23T01:55:44Z-
dc.date.available2016-02-23T01:55:44Z-
dc.date.issued2015-
dc.identifier.citationInternational Journal of Molecular Sciences, 2015, v. 16 n. 10, p. 25067-25079-
dc.identifier.issn1422-0067-
dc.identifier.urihttp://hdl.handle.net/10722/223223-
dc.description.abstractInternal transcribed spacer region (ITS) sequencing is the most extensively used technology for accurate molecular identification of fungal pathogens in clinical microbiology laboratories. Intra-genomic ITS sequence heterogeneity, which makes fungal identification based on direct sequencing of PCR products difficult, has rarely been reported in pathogenic fungi. During the process of performing ITS sequencing on 71 yeast strains isolated from various clinical specimens, direct sequencing of the PCR products showed ambiguous sequences in six of them. After cloning the PCR products into plasmids for sequencing, interpretable sequencing electropherograms could be obtained. For each of the six isolates, 10–49 clones were selected for sequencing and two to seven intra-genomic ITS copies were detected. The identities of these six isolates were confirmed to be <i>Candida glabrata</i> (n = 2), <i>Pichia</i> (<i>Candida</i>) <i>norvegensis</i> (n = 2), <i>Candida tropicalis</i> (n = 1) and <i>Saccharomyces cerevisiae</i> (n = 1). Multiple sequence alignment revealed that one to four intra-genomic ITS polymorphic sites were present in the six isolates, and all these polymorphic sites were located in the ITS1 and/or ITS2 regions. We report and describe the first evidence of intra-genomic ITS sequence heterogeneity in four different pathogenic yeasts, which occurred exclusively in the ITS1 and ITS2 spacer regions for the six isolates in this study.-
dc.languageeng-
dc.publisherMolecular Diversity Preservation International. The Journal's web site is located at http://www.mdpi.org/ijms-
dc.relation.ispartofInternational Journal of Molecular Sciences-
dc.rightsCreative Commons: Attribution 3.0 Hong Kong License-
dc.titleIntra-genomic internal transcribed spacer region sequence heterogeneity and molecular diagnosis in clinical microbiology-
dc.typeArticle-
dc.identifier.emailLau, SKP: skplau@hkucc.hku.hk-
dc.identifier.emailWoo, PCY: pcywoo@hkucc.hku.hk-
dc.identifier.authorityLau, SKP=rp00486-
dc.identifier.authorityWoo, PCY=rp00430-
dc.description.naturepublished_or_final_version-
dc.identifier.doi10.3390/ijms161025067-
dc.identifier.pmid26506340-
dc.identifier.pmcidPMC4632790-
dc.identifier.hkuros256892-
dc.identifier.volume16-
dc.identifier.issue10-
dc.identifier.spage25067-
dc.identifier.epage25079-
dc.publisher.placeSwitzerland-

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