Fast and Accurate Pneumocystis Pneumonia Diagnosis in Human Samples Using a Label-Free Plasmonic Biosensor
Olalla Calvo-Lozano,
Anna Aviñó,
Vicente Friaza,
Alfonso Medina-Escuela,
César S. Huertas,
Enrique J. Calderón,
Ramón Eritja,
Laura M. Lechuga
Affiliations
Olalla Calvo-Lozano
Nanobiosensors and Bioanalytical Applications Group (NanoB2A), Catalan Institute of Nanoscience and Nanotechnology (ICN2), CSIC, CIBER in Bioengineering, Biomaterials and Nanomedicine and BIST, Campus UAB Bellaterra, 08193 Barcelona, Spain
Anna Aviñó
Institute for Advanced Chemistry of Catalonia (IQAC), CSIC, CIBER in Bioengineering, Biomaterials and Nanomedicine c/Jordi Girona 18–26, 08034 Barcelona, Spain
Vicente Friaza
Institute of Biomedicine of Seville (IBiS), University Hospital Virgen del Rocío/CSIC/University of Seville and CIBER in Epidemiology and Public Health, 41013 Seville, Spain
Alfonso Medina-Escuela
Institute for Applied Microelectronics (IUMA), University of Las Palmas de Gran Canaria, 35017 Las Palmas, Spain
César S. Huertas
Nanobiosensors and Bioanalytical Applications Group (NanoB2A), Catalan Institute of Nanoscience and Nanotechnology (ICN2), CSIC, CIBER in Bioengineering, Biomaterials and Nanomedicine and BIST, Campus UAB Bellaterra, 08193 Barcelona, Spain
Enrique J. Calderón
Institute of Biomedicine of Seville (IBiS), University Hospital Virgen del Rocío/CSIC/University of Seville and CIBER in Epidemiology and Public Health, 41013 Seville, Spain
Ramón Eritja
Institute for Advanced Chemistry of Catalonia (IQAC), CSIC, CIBER in Bioengineering, Biomaterials and Nanomedicine c/Jordi Girona 18–26, 08034 Barcelona, Spain
Laura M. Lechuga
Nanobiosensors and Bioanalytical Applications Group (NanoB2A), Catalan Institute of Nanoscience and Nanotechnology (ICN2), CSIC, CIBER in Bioengineering, Biomaterials and Nanomedicine and BIST, Campus UAB Bellaterra, 08193 Barcelona, Spain
Pneumocystis jirovecii is a fungus responsible for human Pneumocystis pneumonia, one of the most severe infections encountered in immunodepressed individuals. The diagnosis of Pneumocystis pneumonia continues to be challenging due to the absence of specific symptoms in infected patients. Moreover, the standard diagnostic method employed for its diagnosis involves mainly PCR-based techniques, which besides being highly specific and sensitive, require specialized personnel and equipment and are time-consuming. Our aim is to demonstrate an optical biosensor methodology based on surface plasmon resonance to perform such diagnostics in an efficient and decentralized scheme. The biosensor methodology employs poly-purine reverse-Hoogsteen hairpin probes for the detection of the mitochondrial large subunit ribosomal RNA (mtLSU rRNA) gene, related to P. jirovecii detection. The biosensor device performs a real-time and label-free identification of the mtLSU rRNA gene with excellent selectivity and reproducibility, achieving limits of detection of around 2.11 nM. A preliminary evaluation of clinical samples showed rapid, label-free and specific identification of P. jirovecii in human lung fluids such as bronchoalveolar lavages or nasopharyngeal aspirates. These results offer a door for the future deployment of a sensitive diagnostic tool for fast, direct and selective detection of Pneumocystis pneumonia disease.
