RESUMO
Exosomes have been widely used in early cancer diagnosis as promising cancer biomarkers due to their abundant tumor-specific molecular information. In this study, we developed a sensitive and straightforward surface-enhanced Raman scattering (SERS) aptasensor to detect exosomes based on gold nanostars-decorated molybdenum disulfide (MoS2) nanocomposites (MoS2-AuNSs). ROX-labeled aptamers (ROX-Apt) were assembled on MoS2-AuNSs surface as recognition probes that specifically bind with transmembrane protein CD63 (a representative surface marker on exosomes). Thus obvious ROX Raman signals were obtained through the synergistic Raman enhancement effect of AuNSs and MoS2 nanosheet. In presence of exosomes, ROX-Apt is preferentially tethered onto exosomes and released from the surface of nanocomposites, resulting in a decrease of the SERS signal. Expectedly, the as-fabricated SERS aptasensor was capable of detecting exosomes in a wide range from 55 to 5.5 × 105 particles µL-1 with a detection limit of 17 particles µL-1. Moreover, the aptasensor exhibited accepted stability and potential clinical applicability.
Assuntos
Aptâmeros de Nucleotídeos , Técnicas Biossensoriais , Exossomos , Nanopartículas Metálicas , Neoplasias Gástricas , Técnicas Biossensoriais/métodos , Ouro , Humanos , Molibdênio , Análise Espectral Raman/métodos , Neoplasias Gástricas/diagnósticoRESUMO
The label-free detection of nucleic acids has attracted interest of scientists due to the fact that it is simple, fast and efficient. Herein, l-cysteine was electropolymerized on the molybdenum disulfide (MoS2) surface to form a stable and electroactive poly(l-cysteine)-functionalized molybdenum disulfide (Pl-Cys/MoS2) sensing interface. Taking microRNA-21 (miRNA-21) as an analytical model, a label-free electrochemical sensor was designed according to the properties of the Pl-Cys/MoS2 sensing interface. Experimental data exhibited that the designed electrochemical sensor exhibited excellent sensitivity, selectivity and stability towards miRNA-21 detection in buffer and real samples. This study offers a methodology to construct a label-free sensing interface by combining MoS2 nanosheets and electroactive molecules.
Assuntos
Técnicas Biossensoriais , MicroRNAs , Cisteína , Dissulfetos , Técnicas Eletroquímicas , MicroRNAs/genética , MolibdênioRESUMO
The state of probe DNA at the biosensing interface greatly affects the detection performance of electrochemical DNA biosensors. Herein, we constructed a target-induced hairpin-mediated biosensing interface to study the effect of probe DNA on the analytical performance of adenosine triphosphate aptamer (ATPA) and adenosine triphosphate (ATP) detection. Moreover, we also explored the electrochemical contribution of the coexisting hairpin and double-stranded DNA (dsDNA) to this sensing interface. Experimental results suggested that the molecular recognition ability and detection performance of the biosensing interface were majorly dependent on the surface density of methylene blue (MB)-labeled probe hairpin DNA and partly affected by the spatial state of the formed dsDNA. When the surface density of hairpin DNA was moderate (5.72 pmol cm-2), this sensing interface determined as low as 0.74 fM ATPA and 5.04 pM ATP with high selectivity and excellent regeneration, respectively. Furthermore, we calculated that the formed dsDNA had a 31.87% contribution in the total electrochemical signal for 10 pM ATPA detection. Based on the above results, we designed an XOR logic gate based on the biosensing interface for ATPA and ATP detection.