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dc.contributor.authorCarvalho-Pereira, Joana
dc.contributor.authorFernandes, Filipa
dc.contributor.authorAraújo, Ricardo
dc.contributor.authorSpringer, Jan
dc.contributor.authorLoeffler, Juergen
dc.contributor.authorBuitrago, Maria Jose 
dc.contributor.authorPais, Célia
dc.contributor.authorSampaio, Paula
dc.identifier.citationJ Fungi (Basel). 2020 Nov 23;6(4):308.es_ES
dc.description.abstractA new and easy polymerase chain reaction (PCR) multiplex strategy, for the identification of the most common fungal species involved in invasive fungal infections (IFI) was developed in this work. Two panels with species-specific markers were designed, the Candida Panel for the identification of Candida species, and the Filamentous Fungi Panel for the identification of Aspergillus species and Rhizopusarrhizus. The method allowed the correct identification of all targeted pathogens using extracted DNA or by colony PCR, showed no cross-reactivity with nontargeted species and allowed identification of different species in mixed infections. Sensitivity reached 10 to 1 pg of DNA and was suitable for clinical samples from sterile sites, with a sensitivity of 89% and specificity of 100%. Overall, the study showed that the new method is suitable for the identification of the ten most important fungal species involved in IFI, not only from positive blood cultures but also from clinical samples from sterile sites. The method provides a unique characteristic, of seeing the peak in the specific region of the panel with the correct fluorescence dye, that aids the ruling out of unspecific amplifications. Furthermore, the panels can be further customized, selecting markers for different species and/or resistance genes.es_ES
dc.description.sponsorshipThis work was supported by the strategic programme UID/BIA/04050/2013 (POCI-01-0145-FEDER-007569) funded by national funds through the FCT I.P., by the ERDF through the COMPETE2020, by the strategic programme UID/BIA/04050/2019 funded by national funds through the FCT I.P., and co-founded by FEDER through Programa Operacional Regional do Norte, in the scope of the project U. Norte Inova. Joana Carvalho-Pereira was funded by FCT/Ministério da Educação e Ciência, grant number SFRH/BD/113384/2015. The authors confirm independence from the sponsors; the content of the article has not been influenced by the sponsors.es_ES
dc.publisherMultidisciplinary Digital Publishing Institute (MDPI) es_ES
dc.subjectAspergillus sp.es_ES
dc.subjectCandida sp.es_ES
dc.subjectRhizopus arrhizuses_ES
dc.subjectFungal infectionses_ES
dc.subjectMolecular diagnosises_ES
dc.subjectMultiplex PCRes_ES
dc.titleMultiplex PCR Based Strategy for Detection of Fungal Pathogen DNA in Patients with Suspected Invasive Fungal Infectionses_ES
dc.typejournal articlees_ES
dc.rights.licenseAtribución 4.0 Internacional*
dc.identifier.journalJournal of fungi (Basel, Switzerland)es_ES
dc.repisalud.centroISCIII::Centro Nacional de Microbiologíaes_ES
dc.rights.accessRightsopen accesses_ES

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Atribución 4.0 Internacional
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