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1.
Biosens Bioelectron ; 261: 116500, 2024 Oct 01.
Artículo en Inglés | MEDLINE | ID: mdl-38896979

RESUMEN

In this work, we present an electrochemical sensor for fast, low-cost, and easy detection of the SARS-CoV-2 spike protein in infected patients. The sensor is based on a selected combination of nanomaterials with a specific purpose. A bioconjugate formed by Few-layer bismuthene nanosheets (FLB) and tetrahedral DNA nanostructures (TDNs) is immobilized on Carbon Screen-Printed Electrodes (CSPE). The TDNs contain on the top vertex an aptamer that specifically binds to the SARS-CoV-2 spike protein, and a thiol group at the three basal vertices to anchor to the FLB. The TDNs are also marked with a redox indicator, Azure A (AA), which allows the direct detection of SARS-CoV-2 spike protein through changes in the current intensity of its electrolysis before and after the biorecognition reaction. The developed sensor can detect SARS-CoV-2 spike protein with a detection limit of 1.74 fg mL-1 directly in nasopharyngeal swab human samples. Therefore, this study offers a new strategy for rapid virus detection since it is versatile enough for different viruses and pathogens.


Asunto(s)
Técnicas Biosensibles , COVID-19 , Límite de Detección , SARS-CoV-2 , Glicoproteína de la Espiga del Coronavirus , SARS-CoV-2/aislamiento & purificación , Técnicas Biosensibles/métodos , Humanos , Glicoproteína de la Espiga del Coronavirus/análisis , Glicoproteína de la Espiga del Coronavirus/química , COVID-19/virología , COVID-19/diagnóstico , Técnicas Electroquímicas/métodos , Nanoestructuras/química , ADN/química , Aptámeros de Nucleótidos/química
2.
Talanta ; 269: 125405, 2024 Mar 01.
Artículo en Inglés | MEDLINE | ID: mdl-37984235

RESUMEN

In this work we describe a highly sensitive method based on a biocatalyzed electrochemiluminescence approach. The system combines, for the first time, the use of few-layer bismuthene (FLB) as a platform for the oriented immobilization of tetrahedral DNA nanostructures (TDNs) specifically designed and synthetized to detect a specific SARS-CoV-2 gene sequence. In one of its vertices, these TDNs contain a DNA capture probe of the open reading frame 1 ab (ORF1ab) of the virus, available for the biorecognition of the target DNA/RNA. At the other three vertices, there are thiol groups that enable the stable anchoring/binding to the FLB surface. This novel geometry/approach enables not only the binding of the TDNs to surfaces, but also the orientation of the capture probe in a direction normal to the bismuthine surface so that it is readily accessible for binding/recognition of the specific SARS-CoV-2 sequence. The analytical signal is based on the anodic electrochemiluminescence (ECL) intensity of luminol which, in turn, arises as a result of the reaction with H2O2, generated by the enzymatic reaction of glucose oxidation, catalyzed by the biocatalytic label avidin-glucose oxidase conjugate (Av-GOx), which acts as co-reactant in the electrochemiluminescent reaction. The method exhibits a limit of detection (LOD) of 4.31 aM and a wide linear range from 14.4 aM to 1.00 µM, and its applicability was confirmed by detecting SARS-CoV-2 in nasopharyngeal samples from COVID-19 patients without the need of any amplification process.


Asunto(s)
Técnicas Biosensibles , Nanoestructuras , Humanos , Peróxido de Hidrógeno/química , Técnicas Biosensibles/métodos , ADN/genética , ADN/química , Nanoestructuras/química , Límite de Detección , Sondas de ADN , Reacción en Cadena de la Polimerasa , Mediciones Luminiscentes/métodos , Técnicas Electroquímicas/métodos
3.
Talanta ; 270: 125497, 2024 Apr 01.
Artículo en Inglés | MEDLINE | ID: mdl-38142611

RESUMEN

In this work we present the preparation of a 2D molybdenum disulphide nanosheets (2D-MoS2) and tetrahedral DNA nanostructures (TDNs) bioconjugate, and its application to the development of a bioassay for rapid and easy virus detection. The bioconjugate has been prepared by using TDNs carrying the capture probe labelled with 6-carboxyfluoresceine (6-FAM). As case of study to assess the utility of the assay developed, we have chosen the SARS-CoV-2 virus. Hence, as probe we have used a DNA sequence complementary to a region of the SARS-CoV-2 ORF1ab gene (TDN-ORF-FAM). This 6-FAM labelled capture probe is located on the top vertex of the tetrahedral DNA nanostructure, the three left vertices of TDNs have a thiol group. These TDNs are bounded to 2D-MoS2 surface through the three thiol groups, allowing the capture probe to be oriented to favour the biorecognition reaction with the analyte. This biorecognition resulting platform has finally been challenged to the detection of the SARS-CoV-2 ORF1ab gene sequence as the target model by measuring fluorescence before and after the hybridization event with a detection limit of 19.7fM. Furthermore, due to high sensitivity of the proposed methodology, it has been applied to directly detect the virus in nasopharyngeal samples of infected patients without the need of any amplification step. The developed bioassay has a wide range of applicability since it can be applied to the detection of any pathogen by changing the probe corresponding to the target sequence. Thus, a novel, hands-on strategy for rapid pathogen detection has proposed and has a high potential application value in the early diagnosis of infections causes by virus or bacteria.


Asunto(s)
Técnicas Biosensibles , Nanoestructuras , Humanos , Molibdeno , ADN/química , Hibridación de Ácido Nucleico , Nanoestructuras/química , Compuestos de Sulfhidrilo , Técnicas Biosensibles/métodos
4.
Med Clin (Barc) ; 136(13): 583-7, 2011 May 14.
Artículo en Español | MEDLINE | ID: mdl-21414644

RESUMEN

BACKGROUND AND OBJECTIVES: Stevens-Johnson syndrome (SJS) and toxic epidermal necrolysis (TEN) are life-threatening drug reactions considered to be part of a single spectrum disease. AIM: The aim of this work is to describe the clinical and epidemiological characteristics of the complex SJS/TEN in a reference University Hospital. PATIENTS AND METHODS: Retrospective study of patients diagnosed with SJS/TEN in a Hospital provided with a reference burn unit (1989-2008). RESULTS: We included 71 patients, 34 SJS and 32 TEN. They presented an average of 62.5% of the body surface area affected and 25.9% of epidermal sloughing. Mucosal involvement was present in all of them. The average of suspected drugs were 2.3 per patient, being painkillers, NSAID and antiepileptic drugs the most frequent. Silver sulfadiazine was the topical treatment most frequently used and 90% received systemic steroids. A fatal outcome was present in 20% of the patients. CONCLUSIONS: The epidemiological and clinical data were consistent with those of other series. Discontinuation of the offending drug, as well as the early admission to a burn unit are the clues for the management of SJS/TEN. Topical treatment with silver sulfadiazine has shown to be useful and safe.


Asunto(s)
Síndrome de Stevens-Johnson/diagnóstico , Adolescente , Adulto , Anciano , Anciano de 80 o más Años , Niño , Preescolar , Femenino , Humanos , Lactante , Masculino , Persona de Mediana Edad , Estudios Retrospectivos , Síndrome de Stevens-Johnson/inducido químicamente , Síndrome de Stevens-Johnson/tratamiento farmacológico , Síndrome de Stevens-Johnson/epidemiología , Adulto Joven
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