Multiplex PCR identification of eight clinically relevant Candida species
Autor(a) principal: | |
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Data de Publicação: | 2007 |
Outros Autores: | , , , , , |
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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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 instacron:RCAAP |
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Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informação |
instacron_str |
RCAAP |
institution |
RCAAP |
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 |
repository.mail.fl_str_mv |
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1799133090784739328 |