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1.
Biosens Bioelectron ; 261: 116500, 2024 Oct 01.
Artigo em Inglês | MEDLINE | ID: mdl-38896979

RESUMO

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.


Assuntos
Técnicas Biossensoriais , COVID-19 , Limite de Detecção , SARS-CoV-2 , Glicoproteína da Espícula de Coronavírus , SARS-CoV-2/isolamento & purificação , Técnicas Biossensoriais/métodos , Humanos , Glicoproteína da Espícula de Coronavírus/análise , Glicoproteína da Espícula de Coronavírus/química , COVID-19/virologia , COVID-19/diagnóstico , Técnicas Eletroquímicas/métodos , Nanoestruturas/química , DNA/química , Aptâmeros de Nucleotídeos/química
2.
Talanta ; 269: 125405, 2024 Mar 01.
Artigo em Inglês | MEDLINE | ID: mdl-37984235

RESUMO

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.


Assuntos
Técnicas Biossensoriais , Nanoestruturas , Humanos , Peróxido de Hidrogênio/química , Técnicas Biossensoriais/métodos , DNA/genética , DNA/química , Nanoestruturas/química , Limite de Detecção , Sondas de DNA , Reação em Cadeia da Polimerase , Medições Luminescentes/métodos , Técnicas Eletroquímicas/métodos
3.
Talanta ; 270: 125497, 2024 Apr 01.
Artigo em Inglês | MEDLINE | ID: mdl-38142611

RESUMO

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.


Assuntos
Técnicas Biossensoriais , Nanoestruturas , Humanos , Molibdênio , DNA/química , Hibridização de Ácido Nucleico , Nanoestruturas/química , Compostos de Sulfidrila , Técnicas Biossensoriais/métodos
4.
Med. clín (Ed. impr.) ; 136(13): 583-587, mayo 2011. ilus, tab
Artigo em Espanhol | IBECS | ID: ibc-89150

RESUMO

Fundamento y objetivo: El síndrome de Stevens-Johnson (SSJ) y la necrólisis epidérmica tóxica (NET) son toxicodermias graves, consideradas formas polares clínico-patológicas de una misma entidad.Objetivo: Describir las características clínicas y epidemiológicas del complejo SSJ/NET en un Hospital Universitario. Pacientes y métodos: Estudio retrospectivo de pacientes diagnosticados de SSJ/NET en un centro que dispone de una Unidad de Quemados (UQ) de referencia (1989-2008). Resultados: Incluimos 71 pacientes (39 SSJ, 32 NET), con una media del 62,5% de superficie corporal total afecta y un 25,9% desepitelizada. El 100% presentaron afectación de mucosas. El número medio de medicamentos sospechosos por paciente fue de 2,3, destacando los antiinflamatorios y anticomiciales. El tratamiento tópico más utilizado fue la sulfadiazina argéntica y se administraron glucocorticoides sistémicos al 90% de pacientes. El 20% fallecieron. Conclusiones: Las características epidemiológicas y clínicas de nuestra serie coinciden con las comunicadas en la literatura. La retirada temprana de los medicamentos sospechosos, el ingreso temprano en una UQ y las curas tópicas con sulfadiazina argéntica son las claves para el manejo del SSJ/NET (AU)


Background and objectives: Stevens-Johnson syndrome (SJS) and toxic epidermal necrolysis (TEN) are lifethreateningdrug 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 (AU)


Assuntos
Humanos , Síndrome de Stevens-Johnson/complicações , Síndrome de Stevens-Johnson/complicações , Estudos Retrospectivos , Sulfadiazina de Prata/uso terapêutico
5.
Med Clin (Barc) ; 136(13): 583-7, 2011 May 14.
Artigo em Espanhol | MEDLINE | ID: mdl-21414644

RESUMO

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.


Assuntos
Síndrome de Stevens-Johnson/diagnóstico , Adolescente , Adulto , Idoso , Idoso de 80 Anos ou mais , Criança , Pré-Escolar , Feminino , Humanos , Lactente , Masculino , Pessoa de Meia-Idade , Estudos Retrospectivos , Síndrome de Stevens-Johnson/induzido quimicamente , Síndrome de Stevens-Johnson/tratamento farmacológico , Síndrome de Stevens-Johnson/epidemiologia , Adulto Jovem
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