Triplex Hybridization-Based Nanosystem for the Rapid Screening of Pneumocystis Pneumonia in Clinical Samples

Pneumocystis pneumonia (PcP) is a disease produced by the opportunistic infection of the fungus Pneumocystis jirovecii. As delayed or unsuitable treatments increase the risk of mortality, the development of rapid and accurate diagnostic tools for PcP are of great importance. Unfortunately, current s...

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Autores: Pla, Luis, Aviñó, Anna, Eritja, Ramón, Ruiz-Gaitán, Alba, Pemán, Javier, Friaza, Vicente, Calderón Sandubete, Enrique José, Santiago-Felipe, Sara
Tipo de recurso: artículo
Estado:Versión publicada
Fecha de publicación:2020
País:España
Institución:Universidad de Sevilla (US)
Repositorio:idUS. Depósito de Investigación de la Universidad de Sevilla
OAI Identifier:oai:idus.us.es:11441/153002
Acceso en línea:https://hdl.handle.net/11441/153002
https://doi.org/10.3390/jof6040292
Access Level:acceso abierto
Palabra clave:Nanoporous anodic alumina
Pneumocystis jirovecii
Molecular gates
Oligonucleotides
Biosensor
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spelling Triplex Hybridization-Based Nanosystem for the Rapid Screening of Pneumocystis Pneumonia in Clinical SamplesPla, LuisAviñó, AnnaEritja, RamónRuiz-Gaitán, AlbaPemán, JavierFriaza, VicenteCalderón Sandubete, Enrique JoséSantiago-Felipe, SaraNanoporous anodic aluminaPneumocystis jiroveciiMolecular gatesOligonucleotidesBiosensorPneumocystis pneumonia (PcP) is a disease produced by the opportunistic infection of the fungus Pneumocystis jirovecii. As delayed or unsuitable treatments increase the risk of mortality, the development of rapid and accurate diagnostic tools for PcP are of great importance. Unfortunately, current standard methods present severe limitations and are far from adequate. In this work, a time-competitive, sensitive and selective biosensor based on DNA-gated nanomaterials for the identification of P. jirovecii is presented. The biosensor consists of a nanoporous anodic alumina (NAA) scaffold which pores are filled with a dye reporter and capped with specific DNA oligonucleotides. In the presence of P. jirovecii genomic DNA, the gated biosensor is open, and the cargo is delivered to the solution where it is monitored through fluorescence spectroscopy. The use of capping oligonucleotides able to form duplex or triplex with P. jirovecii DNA is studied. The final diagnostic tool shows a limit of detection (LOD) of 1 nM of target complementary DNA and does not require previous amplification steps. The method was applied to identify DNA from P. jirovecii in unmodified bronchoalveolar lavage, nasopharyngeal aspirates, and sputum samples in 60 min. This is a promising alternative method for the routinely diagnosis of Pneumocystis pneumonia.MDPIMedicinaCTS-458: Investigación Clínica y Básica en Medicina Interna, Calidad y Continuidad Asistencial2020info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionapplication/pdfapplication/pdfhttps://hdl.handle.net/11441/153002https://doi.org/10.3390/jof6040292reponame:idUS. Depósito de Investigación de la Universidad de Sevillainstname:Universidad de Sevilla (US)InglésJournal of Fungi, 6 (4), 1-14.info:eu-repo/semantics/openAccessoai:idus.us.es:11441/1530022026-06-17T12:51:07Z
dc.title.none.fl_str_mv Triplex Hybridization-Based Nanosystem for the Rapid Screening of Pneumocystis Pneumonia in Clinical Samples
title Triplex Hybridization-Based Nanosystem for the Rapid Screening of Pneumocystis Pneumonia in Clinical Samples
spellingShingle Triplex Hybridization-Based Nanosystem for the Rapid Screening of Pneumocystis Pneumonia in Clinical Samples
Pla, Luis
Nanoporous anodic alumina
Pneumocystis jirovecii
Molecular gates
Oligonucleotides
Biosensor
title_short Triplex Hybridization-Based Nanosystem for the Rapid Screening of Pneumocystis Pneumonia in Clinical Samples
title_full Triplex Hybridization-Based Nanosystem for the Rapid Screening of Pneumocystis Pneumonia in Clinical Samples
title_fullStr Triplex Hybridization-Based Nanosystem for the Rapid Screening of Pneumocystis Pneumonia in Clinical Samples
title_full_unstemmed Triplex Hybridization-Based Nanosystem for the Rapid Screening of Pneumocystis Pneumonia in Clinical Samples
title_sort Triplex Hybridization-Based Nanosystem for the Rapid Screening of Pneumocystis Pneumonia in Clinical Samples
dc.creator.none.fl_str_mv Pla, Luis
Aviñó, Anna
Eritja, Ramón
Ruiz-Gaitán, Alba
Pemán, Javier
Friaza, Vicente
Calderón Sandubete, Enrique José
Santiago-Felipe, Sara
author Pla, Luis
author_facet Pla, Luis
Aviñó, Anna
Eritja, Ramón
Ruiz-Gaitán, Alba
Pemán, Javier
Friaza, Vicente
Calderón Sandubete, Enrique José
Santiago-Felipe, Sara
author_role author
author2 Aviñó, Anna
Eritja, Ramón
Ruiz-Gaitán, Alba
Pemán, Javier
Friaza, Vicente
Calderón Sandubete, Enrique José
Santiago-Felipe, Sara
author2_role author
author
author
author
author
author
author
dc.contributor.none.fl_str_mv Medicina
CTS-458: Investigación Clínica y Básica en Medicina Interna, Calidad y Continuidad Asistencial
dc.subject.none.fl_str_mv Nanoporous anodic alumina
Pneumocystis jirovecii
Molecular gates
Oligonucleotides
Biosensor
topic Nanoporous anodic alumina
Pneumocystis jirovecii
Molecular gates
Oligonucleotides
Biosensor
description Pneumocystis pneumonia (PcP) is a disease produced by the opportunistic infection of the fungus Pneumocystis jirovecii. As delayed or unsuitable treatments increase the risk of mortality, the development of rapid and accurate diagnostic tools for PcP are of great importance. Unfortunately, current standard methods present severe limitations and are far from adequate. In this work, a time-competitive, sensitive and selective biosensor based on DNA-gated nanomaterials for the identification of P. jirovecii is presented. The biosensor consists of a nanoporous anodic alumina (NAA) scaffold which pores are filled with a dye reporter and capped with specific DNA oligonucleotides. In the presence of P. jirovecii genomic DNA, the gated biosensor is open, and the cargo is delivered to the solution where it is monitored through fluorescence spectroscopy. The use of capping oligonucleotides able to form duplex or triplex with P. jirovecii DNA is studied. The final diagnostic tool shows a limit of detection (LOD) of 1 nM of target complementary DNA and does not require previous amplification steps. The method was applied to identify DNA from P. jirovecii in unmodified bronchoalveolar lavage, nasopharyngeal aspirates, and sputum samples in 60 min. This is a promising alternative method for the routinely diagnosis of Pneumocystis pneumonia.
publishDate 2020
dc.date.none.fl_str_mv 2020
dc.type.none.fl_str_mv info:eu-repo/semantics/article
info:eu-repo/semantics/publishedVersion
format article
status_str publishedVersion
dc.identifier.none.fl_str_mv https://hdl.handle.net/11441/153002
https://doi.org/10.3390/jof6040292
url https://hdl.handle.net/11441/153002
https://doi.org/10.3390/jof6040292
dc.language.none.fl_str_mv Inglés
language_invalid_str_mv Inglés
dc.relation.none.fl_str_mv Journal of Fungi, 6 (4), 1-14.
dc.rights.none.fl_str_mv info:eu-repo/semantics/openAccess
eu_rights_str_mv openAccess
dc.format.none.fl_str_mv application/pdf
application/pdf
dc.publisher.none.fl_str_mv MDPI
publisher.none.fl_str_mv MDPI
dc.source.none.fl_str_mv reponame:idUS. Depósito de Investigación de la Universidad de Sevilla
instname:Universidad de Sevilla (US)
instname_str Universidad de Sevilla (US)
reponame_str idUS. Depósito de Investigación de la Universidad de Sevilla
collection idUS. Depósito de Investigación de la Universidad de Sevilla
repository.name.fl_str_mv
repository.mail.fl_str_mv
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