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Article: Internal transcribed spacer region sequence heterogeneity in Rhizopus microsporus: Implications for molecular diagnosis in clinical microbiology laboratories

TitleInternal transcribed spacer region sequence heterogeneity in Rhizopus microsporus: Implications for molecular diagnosis in clinical microbiology laboratories
Authors
KeywordsSpecies Index: Fungi
Mucorales
Rhizopus Microsporus
Issue Date2010
PublisherAmerican Society for Microbiology
Citation
Journal Of Clinical Microbiology, 2010, v. 48 n. 1, p. 208-214 How to Cite?
AbstractAlthough internal transcribed spacer region (ITS) sequence heterogeneity has been reported in a few fungal species, it has very rarely been reported in pathogenic fungi and has never been described in Mucorales, causes of the highly fatal mucormycosis. In a recent outbreak investigation of intestinal mucormycosis due to Rhizopus microsporus infection in patients with hematological malignancies, PCR of the ITS of four of the 28 R. microsporus strains, P11, P12, D3-1, and D4-1, showed thick bands at about 700 bp. Direct sequencing of the purified bands showed frequent double peaks along all of the sequence traces and occasional triple peaks for P12, D3-1, and D4-1. The thick bands of the four R. microsporus strains were purified and cloned. Sequencing of 10 clones for each strain revealed two different ITS sequences for P11 and three different ITS sequences for P12, D3-1, and D4-1. Variations in ITS sequence among the different ribosomal DNA (rDNA) operons in the same strain were observed in only ITS1 and ITS2 and not the 5.8S rDNA region. One copy of P11, P12, and D4-1, respectively, and one copy of P11, P12, D3-1, and D4-1, respectively, showed identical sequences. This represents the first evidence of ITS sequence heterogeneity in Mucorales. ITS sequence heterogeneity is an obstacle to molecular identification and genotyping of fungi in clinical microbiology laboratories. When thick bands and double peaks are observed during PCR sequencing of a gene target, such a strain should be sent to reference laboratories proficient in molecular technologies for further identification and/or genotyping. Copyright © 2010, American Society for Microbiology. All Rights Reserved.
Persistent Identifierhttp://hdl.handle.net/10722/91705
ISSN
2015 Impact Factor: 3.631
2015 SCImago Journal Rankings: 2.151
PubMed Central ID
ISI Accession Number ID
Funding AgencyGrant Number
Consultancy Service for Enhancing Laboratory Surveillance of Emerging Infectious Disease for the Department of Health of the Hong Kong Special Administrative Region of China
Research Grant Council
University Development Fund
Committee for Research and Conference
University of Hong Kong
Funding Information:

This work is partly supported by the Consultancy Service for Enhancing Laboratory Surveillance of Emerging Infectious Disease for the Department of Health of the Hong Kong Special Administrative Region of China, a Research Grant Council Grant, the University Development Fund, a Committee for Research and Conference grant, and an Outstanding Young Researcher Award from The University of Hong Kong.

References

 

DC FieldValueLanguage
dc.contributor.authorWoo, PCYen_HK
dc.contributor.authorLeung, SYen_HK
dc.contributor.authorTo, KKWen_HK
dc.contributor.authorChan, JFWen_HK
dc.contributor.authorNgan, AHYen_HK
dc.contributor.authorCheng, VCCen_HK
dc.contributor.authorLau, SKPen_HK
dc.contributor.authorYuen, KYen_HK
dc.date.accessioned2010-09-17T10:23:51Z-
dc.date.available2010-09-17T10:23:51Z-
dc.date.issued2010en_HK
dc.identifier.citationJournal Of Clinical Microbiology, 2010, v. 48 n. 1, p. 208-214en_HK
dc.identifier.issn0095-1137en_HK
dc.identifier.urihttp://hdl.handle.net/10722/91705-
dc.description.abstractAlthough internal transcribed spacer region (ITS) sequence heterogeneity has been reported in a few fungal species, it has very rarely been reported in pathogenic fungi and has never been described in Mucorales, causes of the highly fatal mucormycosis. In a recent outbreak investigation of intestinal mucormycosis due to Rhizopus microsporus infection in patients with hematological malignancies, PCR of the ITS of four of the 28 R. microsporus strains, P11, P12, D3-1, and D4-1, showed thick bands at about 700 bp. Direct sequencing of the purified bands showed frequent double peaks along all of the sequence traces and occasional triple peaks for P12, D3-1, and D4-1. The thick bands of the four R. microsporus strains were purified and cloned. Sequencing of 10 clones for each strain revealed two different ITS sequences for P11 and three different ITS sequences for P12, D3-1, and D4-1. Variations in ITS sequence among the different ribosomal DNA (rDNA) operons in the same strain were observed in only ITS1 and ITS2 and not the 5.8S rDNA region. One copy of P11, P12, and D4-1, respectively, and one copy of P11, P12, D3-1, and D4-1, respectively, showed identical sequences. This represents the first evidence of ITS sequence heterogeneity in Mucorales. ITS sequence heterogeneity is an obstacle to molecular identification and genotyping of fungi in clinical microbiology laboratories. When thick bands and double peaks are observed during PCR sequencing of a gene target, such a strain should be sent to reference laboratories proficient in molecular technologies for further identification and/or genotyping. Copyright © 2010, American Society for Microbiology. All Rights Reserved.en_HK
dc.languageengen_HK
dc.publisherAmerican Society for Microbiologyen_HK
dc.relation.ispartofJournal of Clinical Microbiologyen_HK
dc.subjectSpecies Index: Fungien_HK
dc.subjectMucoralesen_HK
dc.subjectRhizopus Microsporusen_HK
dc.titleInternal transcribed spacer region sequence heterogeneity in Rhizopus microsporus: Implications for molecular diagnosis in clinical microbiology laboratoriesen_HK
dc.typeArticleen_HK
dc.identifier.emailWoo, PCY: pcywoo@hkucc.hku.hken_HK
dc.identifier.emailTo, KKW: kelvinto@hkucc.hku.hken_HK
dc.identifier.emailChan, JFW: jfwchan@hku.hken_HK
dc.identifier.emailLau, SKP: skplau@hkucc.hku.hken_HK
dc.identifier.emailYuen, KY: kyyuen@hkucc.hku.hken_HK
dc.identifier.authorityWoo, PCY=rp00430en_HK
dc.identifier.authorityTo, KKW=rp01384en_HK
dc.identifier.authorityChan, JFW=rp01736en_HK
dc.identifier.authorityLau, SKP=rp00486en_HK
dc.identifier.authorityYuen, KY=rp00366en_HK
dc.description.naturelink_to_subscribed_fulltext-
dc.identifier.doi10.1128/JCM.01750-09en_HK
dc.identifier.pmid19906897-
dc.identifier.pmcidPMC2812305-
dc.identifier.scopuseid_2-s2.0-73949107807en_HK
dc.identifier.hkuros171984-
dc.relation.referenceshttp://www.scopus.com/mlt/select.url?eid=2-s2.0-73949107807&selection=ref&src=s&origin=recordpageen_HK
dc.identifier.volume48en_HK
dc.identifier.issue1en_HK
dc.identifier.spage208en_HK
dc.identifier.epage214en_HK
dc.identifier.isiWOS:000276151500029-
dc.publisher.placeUnited Statesen_HK
dc.identifier.scopusauthoridWoo, PCY=7201801340en_HK
dc.identifier.scopusauthoridLeung, SY=35299920200en_HK
dc.identifier.scopusauthoridTo, KKW=14323807300en_HK
dc.identifier.scopusauthoridChan, JFW=24278817900en_HK
dc.identifier.scopusauthoridNgan, AHY=14037517900en_HK
dc.identifier.scopusauthoridCheng, VCC=23670479400en_HK
dc.identifier.scopusauthoridLau, SKP=7401596211en_HK
dc.identifier.scopusauthoridYuen, KY=36078079100en_HK

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