Competitive RNA-RNA hybridization-based integrated nanostructured-disposable electrode for highly sensitive determination of miRNAs in cancer cells

Zouari, Mohamed; Campuzano, Susana; Pingarrón, José M.; Raouafi, Noureddine

doi:10.1016/j.bios.2016.12.033

Cited by 57 publications

(34 citation statements)

References 33 publications

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“…Regarding the optimization of the bCp concentration in the DNA sensor (Fig. 2c ), while no significant differences were apparent between the B signals, the S signals increased significantly with the bCp concentration up to 0.1 μM, then decreasing for larger concentrations, which is most likely due to the restricted hybridization efficiency when large amounts of bCp are immobilized 49 , 50 . Results achieved in the optimization of all the other experimental variables are shown in Figure S1 (in the Supplementary Information).…”

Section: Resultsmentioning

confidence: 94%

Electrochemical affinity biosensors for fast detection of gene-specific methylations with no need for bisulfite and amplification treatments

Povedano

Vargas

Montiel

et al. 2018

Sci Rep

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This paper describes two different electrochemical affinity biosensing approaches for the simple, fast and bisulfite and PCR-free quantification of 5-methylated cytosines (5-mC) in DNA using the anti-5-mC antibody as biorecognition element. One of the biosensing approaches used the anti-5-mC as capture bioreceptor and a sandwich type immunoassay, while the other one involved the use of a specific DNA probe and the anti-5-mC as a detector bioreceptor of the captured methylated DNA. Both strategies, named for simplicity in the text as immunosensor and DNA sensor, respectively, were implemented on the surface of magnetic microparticles and the transduction was accomplished by amperometry at screen-printed carbon electrodes by means of the hydrogen peroxide/hydroquinone system. The resulting amperometric biosensors demonstrated reproducibility throughout the entire protocol, sensitive determination with no need for using amplification strategies, and competitiveness with the conventional enzyme-linked immunosorbent assay methodology and the few electrochemical biosensors reported so far in terms of simplicity, sensitivity and assay time. The DNA sensor exhibited higher sensitivity and allowed the detection of the gene-specific methylations conversely to the immunosensor, which detected global DNA methylation. In addition, the DNA sensor demonstrated successful applicability for 1 h-analysis of specific methylation in two relevant tumor suppressor genes in spiked biological fluids and in genomic DNA extracted from human glioblastoma cells.

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Section: Resultsmentioning

confidence: 94%

Electrochemical affinity biosensors for fast detection of gene-specific methylations with no need for bisulfite and amplification treatments

Povedano

Vargas

Montiel

et al. 2018

Sci Rep

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“…MicroRNAs (miRNAs) are recognized as critical regulators of multiple biological processes in cancer, breast cancer in particular, including tumorigenesis, tumor progression, stress response, and drug resistance . Since miRNA discovery, increasing evidence has indicated their essential roles in tumor progression and metastasis; therefore, miRNAs have emerged as potential biomarkers for cancer diagnosis, prognosis, and/or treatment efficacy .…”

Section: Introductionmentioning

confidence: 99%

Quantitative and Specific Detection of Exosomal miRNAs for Accurate Diagnosis of Breast Cancer Using a Surface‐Enhanced Raman Scattering Sensor Based on Plasmonic Head‐Flocked Gold Nanopillars

Lee

Kim

Lee

et al. 2019

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157

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MicroRNAs in exosomes (exosomal miRNAs) have attracted increased attention as cancer biomarkers for early diagnosis and prognosis owing to their stability in body fluids. Since strong association exists between exosomal miRNA expression levels and breast cancer, the development of effective methods that can monitor exosomal miRNA expression both over broad concentration ranges and in ultralow amounts is critical. Here, a surface‐enhanced Raman scattering (SERS)‐based sensing platform is developed for the quantitative determination of exosomal miRNAs. Ultrasensitive exosomal miRNA detection with single‐nucleotide specificity is obtained from enhanced SERS signals from a uniform plasmonic head‐flocked gold nanopillar substrate, which generates multiple hotspots and enables hybridization between short oligonucleotides, i.e., miRNAs and locked nucleic acid probes. The proposed SERS sensor shows an extremely low detection limit without any amplification process, a wide dynamic range (1 am to 100 nm), multiplex sensing capability and sound miRNA recovery in serum. Furthermore, this sensor allows reliable observation of exosomal miRNA expression patterns from breast cancer cell lines and can discriminate breast cancer subtype based on the difference between these patterns. The results suggest that this sensor can be used for universal cancer diagnosis and further biomedical applications through the quantitative measurement of exosomal miRNAs in bodily fluids.

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“…The detection limit (S/N=3) was estimated to be 3.6 fM for miRNA‐21. Compared with the previous reported analysis for miRNA‐21, such as the electrochemical assay (0.086 fM , 84.3 fM ), current detection (25 fM) , impedance detection (60 aM) , and electronic sensor (3 fM) , the well‐designed photoelectrochemical aptasensor exhibited a lower detection limit as well as a wider linear range.…”

Section: Resultsmentioning

confidence: 99%

A Sensitive Photoelectrochemical Aptasensor for miRNA‐21 Based on the Sensitization Effect of CdSe Quantum Dots

et al. 2018

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A novel photoelectrochemical aptasensor for the sensitive detection of miRNA‐21 was designed based on the sensitization effect of CdSe quantum dots as a signal amplification strategy. The TiO2NTs/RGO/AuNPs electrode was prepared by the electrodeposition of graphene oxide and Au nanoparticles on TiO2 nanotubes with electrochemical reduction. The electrode was prepared as the matrix of the electrochemical aptasensor. The hairpin DNA strands decorated with CdSe quantum dots were immobilized on the surface of the electrode by Au−S bonds. Because of the card‐type structure of the hairpin DNA (HP‐DNA), the CdSe quantum dots were close to the base electrode, which would lead to the sensitization effect and cause the increase of the photocurrent intensity. When a certain concentration of the miRNA‐21 was added, miRNA‐21 hybridized with the complementary part of the HP‐DNA. At the same time, the hairpin structure of HP‐DNA was broken into a straight chain structure, so the CdSe quantum dots on the end of the DNA would move away from the electrode surface, the sensitization effect would be weakened, and the photocurrent intensity would be decreased. The designed photoelectrochemical aptasensor exhibited a low detection limit of 5.6 fM and a wide linear range from 20 fM to 0.2 nM, and it also showed good specificity, reproducibility and stability. The signal amplification strategy was also expected to provide a significant photoelectrochemical biosensor platform for the sensitive detection of biomolecules.

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Competitive RNA-RNA hybridization-based integrated nanostructured-disposable electrode for highly sensitive determination of miRNAs in cancer cells

Cited by 57 publications

References 33 publications

Electrochemical affinity biosensors for fast detection of gene-specific methylations with no need for bisulfite and amplification treatments

Electrochemical affinity biosensors for fast detection of gene-specific methylations with no need for bisulfite and amplification treatments

Quantitative and Specific Detection of Exosomal miRNAs for Accurate Diagnosis of Breast Cancer Using a Surface‐Enhanced Raman Scattering Sensor Based on Plasmonic Head‐Flocked Gold Nanopillars

A Sensitive Photoelectrochemical Aptasensor for miRNA‐21 Based on the Sensitization Effect of CdSe Quantum Dots

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