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 Casadellà, Ramón, Ruiz-Gaitán, Alba, Pemán, Javier, Friaza, Javier, Calderón, Enrique J., Aznar, Elena, Martínez-Máñez, Ramón, Santiago-Felipe, Sara
Tipo de documento: artigo
Estado:Versão publicada
Data de publicação:2020
País:España
Recursos:Consejo Superior de Investigaciones Científicas (CSIC)
Repositório:DIGITAL.CSIC. Repositorio Institucional del CSIC
OAI Identifier:oai:digital.csic.es:10261/223149
Acesso em linha:http://hdl.handle.net/10261/223149
Access Level:Acceso aberto
Palavra-chave:Nanoporous anodic alumina
Biosensors
Oligonucleotides
Molecular gates
Pneumocystis jirovecii
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spelling Triplex Hybridization-Based Nanosystem for the Rapid Screening of Pneumocystis Pneumonia in Clinical SamplesPla, LuisAviñó, AnnaEritja Casadellà, RamónRuiz-Gaitán, AlbaPemán, JavierFriaza, JavierCalderón, Enrique J.Aznar, ElenaMartínez-Máñez, RamónSantiago-Felipe, SaraNanoporous anodic aluminaBiosensorsOligonucleotidesMolecular gatesPneumocystis jiroveciiPneumocystis 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.L.P. thanks UPV for her predoctoral fellowship. S.S. thanks the Instituto de Salud Carlos III and the European Social Fund for the financial support “Sara Borrell” (CD16/000237). The authors also thank the Electron Microscopy Service at the UPV and ICTS NANBIOSIS oligonucleotide synthesis platform (U29) for support. The authors would like to thank Jordi Luque Font, from the IRTA Institute of Agrifood Research and Technology-Cabrils, and Josep Armengol from the Instituto Agroforestal Mediterráneo-Universitat Politècnica de València for the supply of T. deformans samples. The fungal culture of S. pompe was supplied by Piet W.J. de Groot from the Regional Center for Biomedical Research, Castilla-La Mancha Science & Technology Park, University of Castilla–La Mancha.Peer reviewedMultidisciplinary Digital Publishing InstituteEritja Casadellà, Ramón [0000-0001-5383-9334]Consejo Superior de Investigaciones Científicas [https://ror.org/02gfc7t72]202020202020info:eu-repo/semantics/articlehttp://purl.org/coar/resource_type/c_6501Publisher's versioninfo:eu-repo/semantics/publishedVersionhttp://hdl.handle.net/10261/223149reponame:DIGITAL.CSIC. Repositorio Institucional del CSICinstname:Consejo Superior de Investigaciones Científicas (CSIC)Ingléshttps://doi.org/10.3390/jof6040292Síinfo:eu-repo/semantics/openAccessoai:digital.csic.es:10261/2231492026-05-22T06:33:51Z
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
Biosensors
Oligonucleotides
Molecular gates
Pneumocystis jirovecii
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 Casadellà, Ramón
Ruiz-Gaitán, Alba
Pemán, Javier
Friaza, Javier
Calderón, Enrique J.
Aznar, Elena
Martínez-Máñez, Ramón
Santiago-Felipe, Sara
author Pla, Luis
author_facet Pla, Luis
Aviñó, Anna
Eritja Casadellà, Ramón
Ruiz-Gaitán, Alba
Pemán, Javier
Friaza, Javier
Calderón, Enrique J.
Aznar, Elena
Martínez-Máñez, Ramón
Santiago-Felipe, Sara
author_role author
author2 Aviñó, Anna
Eritja Casadellà, Ramón
Ruiz-Gaitán, Alba
Pemán, Javier
Friaza, Javier
Calderón, Enrique J.
Aznar, Elena
Martínez-Máñez, Ramón
Santiago-Felipe, Sara
author2_role author
author
author
author
author
author
author
author
author
dc.contributor.none.fl_str_mv Eritja Casadellà, Ramón [0000-0001-5383-9334]
Consejo Superior de Investigaciones Científicas [https://ror.org/02gfc7t72]
dc.subject.none.fl_str_mv Nanoporous anodic alumina
Biosensors
Oligonucleotides
Molecular gates
Pneumocystis jirovecii
topic Nanoporous anodic alumina
Biosensors
Oligonucleotides
Molecular gates
Pneumocystis jirovecii
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
2020
2020
dc.type.none.fl_str_mv info:eu-repo/semantics/article
http://purl.org/coar/resource_type/c_6501
Publisher's version
info:eu-repo/semantics/publishedVersion
format article
status_str publishedVersion
dc.identifier.none.fl_str_mv http://hdl.handle.net/10261/223149
url http://hdl.handle.net/10261/223149
dc.language.none.fl_str_mv Inglés
language_invalid_str_mv Inglés
dc.relation.none.fl_str_mv https://doi.org/10.3390/jof6040292
Sí
dc.rights.none.fl_str_mv info:eu-repo/semantics/openAccess
eu_rights_str_mv openAccess
dc.publisher.none.fl_str_mv Multidisciplinary Digital Publishing Institute
publisher.none.fl_str_mv Multidisciplinary Digital Publishing Institute
dc.source.none.fl_str_mv reponame:DIGITAL.CSIC. Repositorio Institucional del CSIC
instname:Consejo Superior de Investigaciones Científicas (CSIC)
instname_str Consejo Superior de Investigaciones Científicas (CSIC)
reponame_str DIGITAL.CSIC. Repositorio Institucional del CSIC
collection DIGITAL.CSIC. Repositorio Institucional del CSIC
repository.name.fl_str_mv
repository.mail.fl_str_mv
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score 15,812429

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