Proximity sensitive detection of microRNAs using electrochemical impedance spectroscopy biosensors

Roychoudhury, Appan; Dear, James W.; Bachmann, Till T.

doi:10.1016/j.bios.2022.114404

Cited by 22 publications

(12 citation statements)

References 42 publications

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“…Screen-printed gold electrodes (DRP-C223BT, DropSens) were functionalized with PNA probes as per the protocol used in our earlier study. 25 In brief, following electrochemical cleaning using 100 mM sulfuric acid solution and cyclic voltammetry technique (0 to 1.6 V potential range, 100 mV/s scan rate, 10 cycles), the PNA probe molecules were immobilized onto the gold working electrodes by exposing the cleaned electrodes with a mixed solution containing specific concentrations of the probe (thiol-modified PNA probe + 100 μM MCH + 200 μM HDT + 5 mM TCEP in 50% DMSO solution) for 16 h followed by blocking with 1 mM MCH solution for 2 h. Finally, the probe-functionalized electrodes were serially rinsed with 50% (v/v) DMSO aqueous solution and DI water and then used for subsequent impedance measurements (see Figure 1 ).…”

Section: Methodsmentioning

confidence: 99%

Amplification Free Detection of SARS-CoV-2 Using Multi-Valent Binding

et al. 2022

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We present the development of electrochemical impedance spectroscopy (EIS)-based biosensors for sensitive detection of SARS-CoV-2 RNA using multi-valent binding. By increasing the number of probe−target binding events per target molecule, multi-valent binding is a viable strategy for improving the biosensor performance. As EIS can provide sensitive and labelfree measurements of nucleic acid targets during probe−target hybridization, we used multi-valent binding to build EIS biosensors for targeting SARS-CoV-2 RNA. For developing the biosensor, we explored two different approaches including probe combinations that individually bind in a single-valent fashion and the probes that bind in a multi-valent manner on their own. While we found excellent biosensor performance using probe combinations, we also discovered unexpected signal suppression. We explained the signal suppression theoretically using inter-and intra-probe hybridizations which confirmed our experimental findings. With our best probe combination, we achieved a LOD of 182 copies/μL (303 aM) of SARS-CoV-2 RNA and used these for successful evaluation of patient samples for COVID-19 diagnostics. We were also able to show the concept of multi-valent binding with shorter probes in the second approach. Here, a 13-nt-long probe has shown the best performance during SARS-CoV-2 RNA binding. Therefore, multivalent binding approaches using EIS have high utility for direct detection of nucleic acid targets and for point-of-care diagnostics.

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Section: Methodsmentioning

confidence: 99%

Amplification Free Detection of SARS-CoV-2 Using Multi-Valent Binding

et al. 2022

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“…Screen-printed gold electrodes (DRP-C223BT, DropSens) were functionalised with PNA probes according to the protocol used in our earlier study (Roychoudhury et al 2022).…”

Section: Electrode Preparationmentioning

confidence: 99%

“…Electrochemical biosensors are often useful in point-of-care setup due to simple operation and ease in miniaturisation (Roychoudhury et al 2016;Wang 2006). We have previously demonstrated the use of electrochemical impedance spectroscopy (EIS), which is particularly well-suited for developing rapid and sensitive assays for nucleic acid hybridisation-based detection without the need of labelling of targets (Kersaudy-Kerhoas et al 2022;Roychoudhury et al 2022). EIS allows for detailed evaluation of interfacial property changes at the electrode surface caused by the probe modification and subsequent target hybridisation (Lisdat and Schäfer 2008).…”

Section: Introductionmentioning

confidence: 99%

Amplification-free electrochemical biosensor detection of circulating microRNA to identify drug-induced liver injury

Roychoudhury

Dear

Kersaudy-Kerhoas

et al. 2023

Biosensors and Bioelectronics

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“…At present, the most commonly used approaches to miRNA analysis were proposed based on luminescent, , electrochemical, , miRNA-Seq, and quantitative real-time polymerase chain reaction (qPCR) techniques . Nevertheless, no matter what technique, the most important part of a sensor is a signal emitter, which decides the analytical performance.…”

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confidence: 99%

“…4−6 Here, we proposed a new method for measuring miRNA with high sensitivity and selectivity by a dual-mode homogeneous biosensor, which could address the obstacles of miRNAs including the low expression level, few nucleotides, sequence homology, and easy degradation. 7,8 At present, the most commonly used approaches to miRNA analysis were proposed based on luminescent, 9,10 electrochemical, 11,12 miRNA-Seq, 13 and quantitative real-time polymerase chain reaction (qPCR) techniques. 14 Nevertheless, no matter what technique, the most important part of a sensor is a signal emitter, which decides the analytical performance.…”

mentioning

confidence: 99%

Iridium Complex with Specific Intercalation in the G-Quadruplex: A Phosphorescence and Electrochemiluminescence Dual-Mode Homogeneous Biosensor for Enzyme-Free and Label-Free Detection of MicroRNA

Yang

Wang

et al. 2023

ACS Sens.

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Design and exploitation of high-property and multifunctional emitters for achieving immobilization-free, label-free, enzyme-free, and dual-mode determination of microRNA (miRNA) with high sensitivity and selectivity are particularly urgent but remain a huge challenge. Herein, for the first time, we reported an Ir3+-based N-heterocyclic complex, namely, IrPyPt, ascertained its phosphorescence and electrochemiluminescence (ECL) dual-emission behavior in homogeneous solution, and discovered its turn-up phosphorescence and signal-off ECL response to G-quadruplex DNA over other DNAs. Inspired by this, IrPyPt was pioneered as an emitter to combine nucleic acid probes (ON1 and ON2) for developing a phosphorescence and ECL dual-mode homogeneous biosensor, which was applied to label-free, enzyme-free, highly sensitive, and credible analysis of miRNA-21 used as a target analyte. miRNA-21 triggered the hybridization chain reaction of ON1 and ON2 to generate abundant G-quadruplexes, which caught IrPyPt to yield IrPyPt@G-quadruplex, in which the intramolecular rotary motion of IrPyPt was inhibited and IrPyPt was unable to diffuse to the electrode, contributing to a highly boosted phosphorescence emission and a sharply declined ECL emission. With this design, dual-mode analysis of miRNA-21 was achieved with limits of detection down to 1.40 and 0.18 pM, respectively. Furthermore, the dual-mode biosensor was triumphantly applied to determine the concentration of miRNA-21 in cancer cell lysates and thus provided a great potential alternative for miRNA-related diseases’ early and accurate diagnosis.

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Proximity sensitive detection of microRNAs using electrochemical impedance spectroscopy biosensors

Cited by 22 publications

References 42 publications

Amplification Free Detection of SARS-CoV-2 Using Multi-Valent Binding

Amplification Free Detection of SARS-CoV-2 Using Multi-Valent Binding

Amplification-free electrochemical biosensor detection of circulating microRNA to identify drug-induced liver injury

Iridium Complex with Specific Intercalation in the G-Quadruplex: A Phosphorescence and Electrochemiluminescence Dual-Mode Homogeneous Biosensor for Enzyme-Free and Label-Free Detection of MicroRNA

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