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Article

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

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Instituto Interuniversitario de Investigación de Reconocimiento Molecular y Desarrollo Tecnológico, Universitat Politècnica de València, Universitat de València, Camino de Vera s/n, 46022 Valencia, Spain
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Unidad Mixta de Investigación en Nanomedicina y Sensores, Instituto de Investigación Sanitaria La Fe, Universitat Politècnica de València, 46022 Valencia, Spain
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CIBER de Bioingeniería, Biomateriales y Nanomedicina (CIBER-BBN), Spain
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Institute for Advanced Chemistry of Catalonia (IQAC), CSIC, Jordi Girona 18-26, 08034 Barcelona, Spain
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Grupo Acreditado de Infección Grave, Instituto de Investigación Sanitaria La Fe and Servicio de Microbiología, Hospital Universitari i Politècnic La Fe, Avenida Fernando Abril Martorell, 46026 Valencia, Spain
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Instituto de Biomedicina de Sevilla, Hospital Universitario Virgen del Rocío/Consejo Superior de Investigaciones Científicas/Universidad de Sevilla, 41013 Sevilla, Spain
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Centro de Investigación Biomédica en Red de Epidemiología y Salud Pública (CIBERESP), Spain
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Unidad Mixta UPV-CIPF de Investigación en Mecanismos de Enfermedades y Nanomedicina, Universitat Politècnica de València, Centro de Investigación Príncipe Felipe, 46012 Valencia, Spain
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Authors to whom correspondence should be addressed.
J. Fungi 2020, 6(4), 292; https://doi.org/10.3390/jof6040292
Received: 1 October 2020 / Revised: 7 November 2020 / Accepted: 13 November 2020 / Published: 17 November 2020
(This article belongs to the Special Issue Advances in Pneumocystis Infection)
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. View Full-Text
Keywords: nanoporous anodic alumina; Pneumocystis jirovecii; molecular gates; oligonucleotides; biosensor nanoporous anodic alumina; Pneumocystis jirovecii; molecular gates; oligonucleotides; biosensor
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MDPI and ACS Style

Pla, L.; Aviñó, A.; Eritja, R.; Ruiz-Gaitán, A.; Pemán, J.; Friaza, V.; Calderón, E.J.; Aznar, E.; Martínez-Máñez, R.; Santiago-Felipe, S. Triplex Hybridization-Based Nanosystem for the Rapid Screening of Pneumocystis Pneumonia in Clinical Samples. J. Fungi 2020, 6, 292. https://doi.org/10.3390/jof6040292

AMA Style

Pla L, Aviñó A, Eritja R, Ruiz-Gaitán A, Pemán J, Friaza V, Calderón EJ, Aznar E, Martínez-Máñez R, Santiago-Felipe S. Triplex Hybridization-Based Nanosystem for the Rapid Screening of Pneumocystis Pneumonia in Clinical Samples. Journal of Fungi. 2020; 6(4):292. https://doi.org/10.3390/jof6040292

Chicago/Turabian Style

Pla, Luis, Anna Aviñó, Ramón Eritja, Alba Ruiz-Gaitán, Javier Pemán, Vicente Friaza, Enrique J. Calderón, Elena Aznar, Ramón Martínez-Máñez, and Sara Santiago-Felipe. 2020. "Triplex Hybridization-Based Nanosystem for the Rapid Screening of Pneumocystis Pneumonia in Clinical Samples" Journal of Fungi 6, no. 4: 292. https://doi.org/10.3390/jof6040292

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