RESUMEN
The severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), etiological agent for the coronavirus disease 2019 (COVID-19), has resulted in over 775 million global infections. Early diagnosis remains pivotal for effective epidemiological surveillance despite the availability of vaccines. Antigen-based assays are advantageous for early COVID-19 detection due to their simplicity, cost-effectiveness, and suitability for point-of-care testing (PoCT). This study introduces a graphene field-effect transistor-based biosensor designed for high sensitivity and rapid response to the SARS-CoV-2 spike protein. By functionalizing graphene with monoclonal antibodies and applying short-duration gate voltage pulses, we achieve selective detection of the viral spike protein in human serum within 100 µs and at concentrations as low as 1 fg ml-1, equivalent to 8 antigen molecules perµl of blood. Furthermore, the biosensor estimates spike protein concentrations in serum from COVID-19 patients. Our platform demonstrates potential for next-generation PoCT antigen assays, promising fast and sensitive diagnostics for COVID-19 and other infectious diseases.
Asunto(s)
Técnicas Biosensibles , COVID-19 , Grafito , SARS-CoV-2 , Glicoproteína de la Espiga del Coronavirus , Transistores Electrónicos , Glicoproteína de la Espiga del Coronavirus/análisis , Glicoproteína de la Espiga del Coronavirus/inmunología , Técnicas Biosensibles/instrumentación , Técnicas Biosensibles/métodos , Grafito/química , Humanos , SARS-CoV-2/aislamiento & purificación , SARS-CoV-2/inmunología , COVID-19/diagnóstico , COVID-19/sangre , COVID-19/virología , Sensibilidad y Especificidad , Anticuerpos Monoclonales/inmunología , Anticuerpos Monoclonales/químicaRESUMEN
Manufactured titanium dioxide nanoparticles (TiO2-NP) have been intensely applied in numerous industrial products and may be a risk for aquatic systems as they are not completely removed from domestic and industrial wastes after water treatment. This study evaluated the osmo- and ionic balance, Na+/K+-ATPase, H+-ATPase and carbonic anhydrase activities and the mitochondria-rich cells (MRC) in the gills and kidney of the Neotropical fish Prochilodus lineatus after 2 (acute) and 14 (subchronic) days of exposure to nominal 0, 1, 5, 10 and 50â¯mg L-1 TiO2-NP. The nominal concentrations corresponded to 0.0, 0.6, 1.6, 2.7 and 18.1â¯mg L-1 suspended TiO2-NP, respectively, in the water column one hour after NP introduction and were maintained for at least 24â¯h. Acute exposure to TiO2-NP decreased plasma osmolality and Ca2+ levels. Na+/K+-ATPase, H+-ATPase and carbonic anhydrase activities were inhibited in the gills, but not in the kidney. Total MRC density did not change in gills and kidneys. At gill surface, total MRC density decreased in fish exposed to 50â¯mg L-1 TiO2-NP and the total MRC fractional surface area unchanged although, there were some changes in the fractional area of MRC with apical microvilli (MRCm) and MRC with apical sponge-like structure (MRCs). MRCm was more abundant than MRCs. After subchronic exposure, there was no change in plasma osmolality, ionic balance and enzyme activities. Total gill MRC density increased in the filament epithelium and renal tubules. In the gills, MRC contacting water exhibited some adjustments. Total MRC and fractional surface area unchanged, but there was an increase of MRCs contacting water at gill surface after exposure to10 and 50â¯mg L-1 TiO2-NP. MRC proliferation in filament epithelium and in renal tubules as well as the increasing MRCs at gill surface may have contributed to avoid change in plasma osmolality, ionic balance and enzyme activities and suggested a cellular physiological and morphological response to restore and maintain osmotic and ionic homeostasis after subchronic exposure.
Asunto(s)
Characiformes/metabolismo , Mitocondrias/metabolismo , Nanopartículas/toxicidad , Titanio/toxicidad , Clima Tropical , Animales , Anhidrasas Carbónicas/metabolismo , Branquias/metabolismo , Branquias/ultraestructura , Iones , Riñón/metabolismo , Mitocondrias/efectos de los fármacos , Mitocondrias/ultraestructura , Concentración Osmolar , Tamaño de la Partícula , ATPasas de Translocación de Protón/metabolismo , ATPasa Intercambiadora de Sodio-Potasio/metabolismo , Agua/química , Contaminantes Químicos del Agua/toxicidad , Equilibrio HidroelectrolíticoRESUMEN
Graphene is a material that has attracted attention with regard to sensing and biosensing applications in recent years. Here, we report a novel treatment (using ultrasonic bath and ultrasonic tip) to obtain graphene oxide (GO) and a new stable conducting film using reduced graphene oxide (RGO) and dihexadecylphosphate film (DHP). The GO was obtained by chemical exfoliation and it was reduced using NaBH4. Subsequently, RGO-DHP dispersion was prepared and it was dropped onto a glassy carbon electrode by casting technique. The electrode was characterized by cyclic voltammetry and electrochemical spectroscopy impedance. The voltammetric behavior of the RGO-DHP/GC electrode in the presence of estradiol was studied, and the results reported an irreversible oxidation peak current at 0.6V. Under the optimal experimental conditions, using linear sweep adsorptive stripping voltammetry, the detection limit obtained for this hormone was 7.7×10(-8)mol L(-1). The proposed electrode can be attractive for applications as electrochemical sensors and biosensors.