RESUMO
A novel electrochemical nanobiosensor for the detection of miR-155 (as breast cancer biomarker) is introduced . Fe3O4NPs@Ag core-shell nanoparticles were synthesized and their shape and characteristics were confirmed by scanning electron microscope (SEM) imaging, Fourier-transform infrared spectroscopy (FTIR), and X-ray diffraction (XRD) methods. Synthesized nanoparticles were applied onto the magnetic bar carbon paste electrode and then the amine-modified anti-miR-155 (single-stranded probes) was applied on the modified electrode surface and upon hybridization with target miR-155, resveratrol (RSV) was eventually applied as an electrochemical label on the double-strand oligonucleotide. Differential pulse voltammetry (DPV) of the oxidation peak of RSV was assumed as the final signal by sweeping potential from 0 to 0.6 V (vs. Ag/AgCl). The fabrication process was optimized through a series of experiments and the optimized process was confirmed using cyclic voltammetry (CV) and electrochemical impedance spectroscopy (EIS). The linear range of the fabricated nanobiosensor was 0.5 fM to 1.0 nM and the detection limit was 0.15 fM. The nanobiosensor was able to pass reproducibility and specificity tests using different types of mismatched target sequences.Spiked real samples of human serum were used to confirm that the nanobiosensor enables detection of miR-155 without any significant interferences from other moieties and molecules. Finally, the molecular dynamics simulation of the RSV interaction with single- and double-stranded oligonucleotide was performed and confirmed the preferential binding of RSV to double-stranded DNA; therefore, it can be used as the electrochemical label of DNA and/or miRNA hybridization-based biosensors. Graphical abstract.
Assuntos
Técnicas Biossensoriais/métodos , DNA de Cadeia Simples/química , Nanopartículas de Magnetita/química , MicroRNAs/sangue , Oligodesoxirribonucleotídeos/química , Resveratrol/química , Biomarcadores Tumorais/sangue , Biomarcadores Tumorais/genética , DNA de Cadeia Simples/genética , DNA de Cadeia Simples/metabolismo , Técnicas Eletroquímicas/métodos , Humanos , Ácidos Nucleicos Imobilizados/química , Ácidos Nucleicos Imobilizados/genética , Ácidos Nucleicos Imobilizados/metabolismo , Limite de Detecção , MicroRNAs/genética , Simulação de Acoplamento Molecular , Nanocompostos/química , Hibridização de Ácido Nucleico , Oligodesoxirribonucleotídeos/genética , Oligodesoxirribonucleotídeos/metabolismo , Reprodutibilidade dos Testes , Resveratrol/metabolismo , Prata/químicaRESUMO
Acute lymphoblastic leukemia (ALL) is one of the most prevalent cancers in children and microRNA-128 is amongst the most useful biomarkers not only for diagnosis of ALL, but also for discriminating ALL from acute myeloid leukemia (AML). In this study, a novel electrochemical nanobiosensor based on reduced graphene oxide (RGO) and gold nanoparticles (AuNPs) has been fabricated to detect miRNA-128. Cyclic Voltametery (CV), Square Wave Voltametery (SWV) and Electrochemical Impedance Spectroscopy (EIS) have been applied to characterize the nanobiosensor. Hexacyanoferrate as a label-free and methylene blue as a labeling material were used in the design of the nanobiosensors. It was found that the modified electrode has excellent selectivity and sensitivity to miR-128, with a limit of detection of 0.08761 fM in label-free and 0.00956 fM in labeling assay. Additionally, the examination of real serum samples of ALL and AML patients and control cases confirms that the designed nanobiosensor has the potential to detect and discriminate these two cancers and the control samples.
Assuntos
Técnicas Biossensoriais , Nanopartículas Metálicas , MicroRNAs , Criança , Humanos , Ouro/química , Técnicas Eletroquímicas/métodos , Limite de Detecção , Nanopartículas Metálicas/química , Técnicas Biossensoriais/métodosRESUMO
Nowadays, diagnosing early-stage cancers can be vital for saving patients and dramatically decreases mortality rates. Therefore, specificity and sensitivity in the detection of cancer antigens should be elaborately ensured. Some early-stage cancers can be diagnosed via detecting the cancer antigen CA-125, such as ovarian cancer, and required treatments can be applied more efficiently. Thus, detection of CA-125 by employing various optical or electrochemical biosensors is a preliminary and crucial step to treating cancers. In this review, a diverse range of optical and electrochemical means of detecting CA-125 are reviewed. Furthermore, an applicable comparison of their performance and sensitivity is provided, several commercial detection kits are investigated, and their applications are compared and discussed to determine whether they are applicable and accurate enough.
Assuntos
Técnicas Biossensoriais , Neoplasias Ovarianas , Feminino , Humanos , Antígeno Ca-125RESUMO
The widespread accessibility of commercial/clinically-viable electrochemical diagnostic systems for rapid quantification of viral proteins demands translational/preclinical investigations. Here, Covid-Sense (CoVSense) antigen testing platform; an all-in-one electrochemical nano-immunosensor for sample-to-result, self-validated, and accurate quantification of the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) nucleocapsid (N)-proteins in clinical examinations is developed. The platform's sensing strips benefit from a highly-sensitive, nanostructured surface, created through the incorporation of carboxyl-functionalized graphene nanosheets, and poly(3,4-ethylenedioxythiophene) polystyrene sulfonate (PEDOT:PSS) conductive polymers, enhancing the overall conductivity of the system. The nanoengineered surface chemistry allows for compatible direct assembly of bioreceptor molecules. CoVSense offers an inexpensive (<$2 kit) and fast/digital response (<10 min), measured using a customized hand-held reader (<$25), enabling data-driven outbreak management. The sensor shows 95% clinical sensitivity and 100% specificity (Ct<25), and overall sensitivity of 91% for combined symptomatic/asymptomatic cohort with wildtype SARS-CoV-2 or B.1.1.7 variant (N = 105, nasal/throat samples). The sensor correlates the N-protein levels to viral load, detecting high Ct values of ≈35, with no sample preparation steps, while outperforming the commercial rapid antigen tests. The current translational technology fills the gap in the workflow of rapid, point-of-care, and accurate diagnosis of COVID-19.
Assuntos
COVID-19 , SARS-CoV-2 , Humanos , COVID-19/diagnóstico , Sensibilidade e Especificidade , Nucleocapsídeo , AntígenosRESUMO
The detection of nucleic acids and their mutation derivatives is vital for biomedical science and applications. Although many nucleic acid biosensors have been developed, they often require pretreatment processes, such as target amplification and tagging probes to nucleic acids. Moreover, current biosensors typically cannot detect sequence-specific mutations in the targeted nucleic acids. To address the above problems, herein, we developed an electrochemical nanobiosensing system using a phenomenon comprising metal ion intercalation into the targeted mismatched double-stranded nucleic acids and a homogeneous Au nanoporous electrode array (Au NPEA) to obtain (i) sensitive detection of viral RNA without conventional tagging and amplifying processes, (ii) determination of viral mutation occurrence in a simple detection manner, and (iii) multiplexed detection of several RNA targets simultaneously. As a proof-of-concept demonstration, a SARS-CoV-2 viral RNA and its mutation derivative were used in this study. Our developed nanobiosensor exhibited highly sensitive detection of SARS-CoV-2 RNA (â¼1 fM detection limit) without tagging and amplifying steps. In addition, a single point mutation of SARS-CoV-2 RNA was detected in a one-step analysis. Furthermore, multiplexed detection of several SARS-CoV-2 RNAs was successfully demonstrated using a single chip with four combinatorial NPEAs generated by a 3D printing technique. Collectively, our developed nanobiosensor provides a promising platform technology capable of detecting various nucleic acids and their mutation derivatives in highly sensitive, simple, and time-effective manners for point-of-care biosensing.
Assuntos
Técnicas Biossensoriais , COVID-19 , Nanoporos , Ácidos Nucleicos , Humanos , RNA Viral/genética , Técnicas Eletroquímicas/métodos , Nucleotídeos , SARS-CoV-2 , Eletrodos , Técnicas Biossensoriais/métodos , Técnicas de Amplificação de Ácido Nucleico/métodosRESUMO
Cancer is one of the most important causes of mortality in the world, which can be severely reduced by early detection to avoid future problems in the field of economics and mental health. Hence, electrochemical nanobiosensors as portable devices for rapid detection of  cancer biomarkers, have found an important place in clinical medicine for diagnosis, managements or cancer screening. Although, these biosensors have been receiving attention in the recent years, their principles are unchanged. By progress in nanotechnology, a great potential has been giving to nanobiosensors. Applications of a wide variety of nanomaterials in developing electrochemical biosensors, led to the production of potential nanobiosensors. Due to the high electrical conductivity, and increased surface area relative to the volume along with more repeatability, the application of NPs in electrochemical biosensors has been developed. Therefore, in this review, we discussed the impact of nanomaterials on the accuracy of biosensors in early cancer detection such as lung, prostate, breast, and other cancers. However, the modification of electrode performance by nanomaterials is relatively complicated, which causes limitation for some nanomaterials to be used inbiosensor applications. Indeed, the construction of electrodes based on nanomaterial requires a simple, reliable and inexpensive route to increase the sensitivity and reproducibility. Thus, the aim of this study can be defined as determining the detection limit of electrochemical nanobiosensors as well as introducing the challenges of fabricating and designing electrochemical nanobiosensors based on nanomaterials and their evaluations in the future medical setting.