RESUMO
A simple carbon nanodot-based electrogenerated chemiluminescence biosensor is described for sensitive and selective detection of microRNA-21 (miRNA-21), a biomarker of several pathologies including cardiovascular diseases (CVDs). The photoluminescent carbon nanodots (CNDs) were obtained using a new synthesis method, simply by treating tiger nut milk in a microwave reactor. The synthesis is environmentally friendly, simple, and efficient. The optical properties and morphological characteristics of the CNDs were exhaustively investigated, confirming that they have oxygen and nitrogen functional groups on their surfaces and exhibit excitation-dependent fluorescence emission, as well as photostability. They act as co-reactant agents in the anodic electrochemiluminescence (ECL) of [Ru(bpy)3]2+, producing different signals for the probe (single-stranded DNA) and the hybridized target (double-stranded DNA). These results paved the way for the development of a sensitive ECL biosensor for the detection of miRNA-21. This was developed by immobilization of a thiolated oligonucleotide, fully complementary to the miRNA-21 sequence, on the disposable gold electrode. The target miRNA-21 was hybridized with the probe on the electrode surface, and the hybridization was detected by the enhancement of the [Ru(bpy)3]2+/DNA ECL signal using CNDs. The biosensor shows a linear response to miRNA-21 concentration up to 100.0 pM with a detection limit of 0.721 fM. The method does not require complex labeling steps, and has a rapid response. It was successfully used to detect miRNA-21 directly in serum samples from heart failure patients without previous RNA extraction neither amplification process.
Assuntos
Técnicas Biossensoriais/métodos , Técnicas Eletroquímicas/métodos , Substâncias Luminescentes/química , Medições Luminescentes/métodos , MicroRNAs/sangue , Pontos Quânticos/química , Técnicas Biossensoriais/instrumentação , Carbono/química , Complexos de Coordenação/química , Técnicas Eletroquímicas/instrumentação , Eletrodos , Ouro/química , Insuficiência Cardíaca/sangue , Humanos , Ácidos Nucleicos Imobilizados/genética , Limite de Detecção , Medições Luminescentes/instrumentação , Masculino , MicroRNAs/genética , Hibridização de Ácido Nucleico , Oligodesoxirribonucleotídeos/química , Oligodesoxirribonucleotídeos/genética , Compostos de Rutênio/químicaRESUMO
A novel immunosensor based on electrochemiluminescence resonance energy transfer (ECL-RET) for the sensitive determination of N protein of the SARS-CoV-2 coronavirus is described. For this purpose, bifunctional core@shell nanoparticles composed of a Pt-coated Au core and finally decorated with small Au inlays (Au@Pt/Au NPs) have been synthesized to act as ECL acceptor, using [Ru (bpy)3]2+ as ECL donor. These nanoparticles are efficient signaling probes in the immunosensor developed. The proposed ECL-RET immunosensor has a wide linear response to the concentration of N protein of the SARS-CoV-2 coronavirus with a detection limit of 1.27 pg/mL. Moreover, it has a high stability and shows no response to other proteins related to different virus. The immunosensor has achieved the quantification of N protein of the SARS-CoV-2 coronavirus in saliva samples. Results are consistent with those provided by a commercial colorimetric ELISA kit. Therefore, the developed immunosensor provides a feasible and reliable tool for early and effective detection of the virus to protect the population.
Assuntos
Técnicas Biossensoriais , COVID-19 , Nanopartículas Metálicas , Humanos , Ouro , SARS-CoV-2 , Medições Luminescentes/métodos , Técnicas Biossensoriais/métodos , Imunoensaio/métodos , COVID-19/diagnóstico , Técnicas Eletroquímicas/métodos , Limite de DetecçãoRESUMO
Given the great utility that having fast, efficient and cost-effective methods for the detection of SARS-CoV-2 in wastewater can have in controlling the pandemic caused by this virus, the development of new dependable and specific SARS-CoV-2 coronavirus sensing devices to be applied to wastewater is essential to promote public health interventions. Therefore, herein we propose a new method to detect SARS-CoV-2 in wastewater based on a carbon nanodots-amplified electrochemiluminescence immunosensor for the determination of the SARS-CoV-2 Spike S1 protein. For the construction of the immunosensor, N-rich carbon nanodots have been synthetized with a double function: to contribute as amplifiers of the electrochemiluminescent signal in presence of [Ru(bpy)3]2+ and as antibody supports by providing functional groups capable of covalently interacting with the SARS-CoV-2 Spike S1 antibody. The proposed ECL immunosensor has demonstrated a high specificity in presence of other virus-related proteins and responded linearly to SARS-CoV-2 Spike S1 concentration over a wide range with a limit of detection of 1.2 pg/mL. The immunosensor has an excellent stability and achieved the detection of SARS-CoV-2 Spike S1 in river and urban wastewater, which supplies a feasible and reliable sensing platform for early virus detection and therefore to protect the population. The detection of SARS-CoV-2 Spike S1 in urban wastewater can be used as a tool to measure the circulation of the virus in the population and to detect a possible resurgence of COVID-19.
Assuntos
Técnicas Biossensoriais , COVID-19 , Técnicas Biossensoriais/métodos , COVID-19/diagnóstico , Carbono , Humanos , Imunoensaio/métodos , SARS-CoV-2 , Águas ResiduáriasRESUMO
A novel electrochemiluminescence (ECL) sensor for the sensitive detection of taurine was developed. Taurine contains an aliphatic amine that gives it co-reactant properties. The ECL response of the taurine/[Ru(bpy)3]2+ system was analyzed on two different electrodes surfaces, screen-printed graphene and gold electrodes, before and after modification with ZnO nanowires (ZnO NWs). The ZnO NWs modified electrode yielded an enhanced ECL signal, allowing rapid detection of taurine at 5.5â¯×â¯10-6â¯molâ¯L-1 detection limit. The ECL signal is stable and reproducible. The sensor has been applied to the determination of taurine in a commercial taurine supplement.