Multiplex PCR identification of eight clinically relevant Candida species

Detalhes bibliográficos
Autor(a) principal: Carvalho, Agostinho
Data de Publicação: 2007
Outros Autores: Oliveira, Sofia Costa de, Martins, M. L., Pina-Vaz, C., Rodrigues, A. G., Ludovico, Paula, Rodrigues, Fernando José dos Santos
Tipo de documento: Artigo
Idioma: eng
Título da fonte: Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)
Texto Completo: http://hdl.handle.net/1822/67702
Resumo: Invasive fungal infections, specifically candidemia, constitute major public health problems with high mortality rates. Therefore, in the last few years, the development of novel diagnostic methods has been considered a critical issue. Herein we describe a multiplex PCR strategy allowing the identification of 8 clinically relevant yeasts of the Candida genus, namely C. albicans, C. glabrata, C. parapsilosis, C. tropicalis, C. krusei, C. guilliermondii, C. lusitaniae and C. dubliniensis. This method is based on the amplification of two fragments from the ITS1 and ITS2 regions by the combination of 2 yeast-specific and 8 species-specific primers in a single PCR reaction. Results from the identification of 231 clinical isolates are presented pointing to the high specificity of this procedure. Furthermore, several Candida isolates were identified directly from clinical specimens which also attests to the method's direct laboratory application. The results from the multiplex reactions with other microorganisms that usually co-infect patients also confirmed its high specificity in the identification of Candida species. Moreover, this method is simple and presents a sensitivity of approximately 2 cells per ml within 5 hours. Furthermore, it allows discrimination of individual Candida species within polyfungal samples. This novel method may therefore provide a clinical diagnostic procedure with direct applicability.
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spelling Multiplex PCR identification of eight clinically relevant Candida speciesCandidaCandidiasisDNA PrimersDNA, Ribosomal SpacerHumansPolymerase Chain ReactionSensitivity and SpecificityCandidemiaDiagnosticIdentificationPCRMultiplexScience & TechnologyInvasive fungal infections, specifically candidemia, constitute major public health problems with high mortality rates. Therefore, in the last few years, the development of novel diagnostic methods has been considered a critical issue. Herein we describe a multiplex PCR strategy allowing the identification of 8 clinically relevant yeasts of the Candida genus, namely C. albicans, C. glabrata, C. parapsilosis, C. tropicalis, C. krusei, C. guilliermondii, C. lusitaniae and C. dubliniensis. This method is based on the amplification of two fragments from the ITS1 and ITS2 regions by the combination of 2 yeast-specific and 8 species-specific primers in a single PCR reaction. Results from the identification of 231 clinical isolates are presented pointing to the high specificity of this procedure. Furthermore, several Candida isolates were identified directly from clinical specimens which also attests to the method's direct laboratory application. The results from the multiplex reactions with other microorganisms that usually co-infect patients also confirmed its high specificity in the identification of Candida species. Moreover, this method is simple and presents a sensitivity of approximately 2 cells per ml within 5 hours. Furthermore, it allows discrimination of individual Candida species within polyfungal samples. This novel method may therefore provide a clinical diagnostic procedure with direct applicability.Agostinho Carvalho was financially supported by a fellowship from Fundação para a Ciência e Tecnologia, Portugal (contract SFRH/BD/11837/2003). This study was supported by Fundação para a Ciência e Tecnologia, Portugal (POCI/SAU-ESP/61080/2004).Oxford University PressUniversidade do MinhoCarvalho, AgostinhoOliveira, Sofia Costa deMartins, M. L.Pina-Vaz, C.Rodrigues, A. G.Ludovico, PaulaRodrigues, Fernando José dos Santos2007-112007-11-01T00:00:00Zinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/articleapplication/pdfhttp://hdl.handle.net/1822/67702eng1369-37861460-270910.1080/1369378070150178717885953info:eu-repo/semantics/openAccessreponame:Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)instname:Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informaçãoinstacron:RCAAP2023-07-21T12:51:36Zoai:repositorium.sdum.uminho.pt:1822/67702Portal AgregadorONGhttps://www.rcaap.pt/oai/openaireopendoar:71602024-03-19T19:50:31.770583Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) - Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informaçãofalse
dc.title.none.fl_str_mv Multiplex PCR identification of eight clinically relevant Candida species
title Multiplex PCR identification of eight clinically relevant Candida species
spellingShingle Multiplex PCR identification of eight clinically relevant Candida species
Carvalho, Agostinho
Candida
Candidiasis
DNA Primers
DNA, Ribosomal Spacer
Humans
Polymerase Chain Reaction
Sensitivity and Specificity
Candidemia
Diagnostic
Identification
PCR
Multiplex
Science & Technology
title_short Multiplex PCR identification of eight clinically relevant Candida species
title_full Multiplex PCR identification of eight clinically relevant Candida species
title_fullStr Multiplex PCR identification of eight clinically relevant Candida species
title_full_unstemmed Multiplex PCR identification of eight clinically relevant Candida species
title_sort Multiplex PCR identification of eight clinically relevant Candida species
author Carvalho, Agostinho
author_facet Carvalho, Agostinho
Oliveira, Sofia Costa de
Martins, M. L.
Pina-Vaz, C.
Rodrigues, A. G.
Ludovico, Paula
Rodrigues, Fernando José dos Santos
author_role author
author2 Oliveira, Sofia Costa de
Martins, M. L.
Pina-Vaz, C.
Rodrigues, A. G.
Ludovico, Paula
Rodrigues, Fernando José dos Santos
author2_role author
author
author
author
author
author
dc.contributor.none.fl_str_mv Universidade do Minho
dc.contributor.author.fl_str_mv Carvalho, Agostinho
Oliveira, Sofia Costa de
Martins, M. L.
Pina-Vaz, C.
Rodrigues, A. G.
Ludovico, Paula
Rodrigues, Fernando José dos Santos
dc.subject.por.fl_str_mv Candida
Candidiasis
DNA Primers
DNA, Ribosomal Spacer
Humans
Polymerase Chain Reaction
Sensitivity and Specificity
Candidemia
Diagnostic
Identification
PCR
Multiplex
Science & Technology
topic Candida
Candidiasis
DNA Primers
DNA, Ribosomal Spacer
Humans
Polymerase Chain Reaction
Sensitivity and Specificity
Candidemia
Diagnostic
Identification
PCR
Multiplex
Science & Technology
description Invasive fungal infections, specifically candidemia, constitute major public health problems with high mortality rates. Therefore, in the last few years, the development of novel diagnostic methods has been considered a critical issue. Herein we describe a multiplex PCR strategy allowing the identification of 8 clinically relevant yeasts of the Candida genus, namely C. albicans, C. glabrata, C. parapsilosis, C. tropicalis, C. krusei, C. guilliermondii, C. lusitaniae and C. dubliniensis. This method is based on the amplification of two fragments from the ITS1 and ITS2 regions by the combination of 2 yeast-specific and 8 species-specific primers in a single PCR reaction. Results from the identification of 231 clinical isolates are presented pointing to the high specificity of this procedure. Furthermore, several Candida isolates were identified directly from clinical specimens which also attests to the method's direct laboratory application. The results from the multiplex reactions with other microorganisms that usually co-infect patients also confirmed its high specificity in the identification of Candida species. Moreover, this method is simple and presents a sensitivity of approximately 2 cells per ml within 5 hours. Furthermore, it allows discrimination of individual Candida species within polyfungal samples. This novel method may therefore provide a clinical diagnostic procedure with direct applicability.
publishDate 2007
dc.date.none.fl_str_mv 2007-11
2007-11-01T00:00:00Z
dc.type.status.fl_str_mv info:eu-repo/semantics/publishedVersion
dc.type.driver.fl_str_mv info:eu-repo/semantics/article
format article
status_str publishedVersion
dc.identifier.uri.fl_str_mv http://hdl.handle.net/1822/67702
url http://hdl.handle.net/1822/67702
dc.language.iso.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv 1369-3786
1460-2709
10.1080/13693780701501787
17885953
dc.rights.driver.fl_str_mv info:eu-repo/semantics/openAccess
eu_rights_str_mv openAccess
dc.format.none.fl_str_mv application/pdf
dc.publisher.none.fl_str_mv Oxford University Press
publisher.none.fl_str_mv Oxford University Press
dc.source.none.fl_str_mv reponame:Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)
instname:Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informação
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reponame_str Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)
collection Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)
repository.name.fl_str_mv Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) - Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informação
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