Detection of a few DNA copies by real-time electrochemical polymerase chain reaction

Moreau, Myriam; Delile, Sébastien; Sharma, Ashwani; Fave, Claire; Perrier, Aurélie; Limoges, Benoı̂t; Marchal, Damien

doi:10.1039/c7an00978j

Cited by 11 publications

(7 citation statements)

References 31 publications

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“…We propose ultra-low cost portable (palm size device) electrochemical based method using the Electrode DNA Chip and based on principle of electrical conductance of DNA/RNA hybrid. The proposed method is modification of the method developed earlier [21].…”

Section: Introductionmentioning

confidence: 99%

Computational Intelligence Methods for the Diagnosis of COVID-19

Akermi¹,

Sinha

Johari

et al. 2020

Studies in Computational Intelligence

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COVID-19 is a global catastrophe affecting over 200 nations worldwide. The RT-PCR and antibody based detection assay are considered as a standard diagnostic method and are expensive, time-consuming with low sensitivity (40-80%) making it imperative to look for alternatives. COVID-19 patients display severe lung damage which can be spotted in chest X-ray and CT scan which can be analyzed by computational methods for confirmation of the disease. Apart from above techniques, we have discussed battery of computational intelligence tools like docking, molecular dynamics and quantum mechanics (QM), the novel biosensor based diagnostic tool has been developed. The biosensor involves hybridization of SARS-CoV-2 RNA with cDNA to form RNA (viral)-cDNA (probe) hybrid, with intercalation of transition metal Osmium-Ruthenium (II) redox probe that release electrons. The electrons generated by redox metal measured by respective electrodes and detect even miniscule quantities of viral-RNA in the sample. The computational methods support efficient measurement of interaction of redox probe with viral genetic material and intercalation of the Osmium-Ruthenium (II) redox probe between RNA-cDNA hybrid. The computational docking offers proof of concept with better sensitivity, speed and

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Section: Introductionmentioning

confidence: 99%

Computational Intelligence Methods for the Diagnosis of COVID-19

Akermi¹,

Sinha

Johari

et al. 2020

Studies in Computational Intelligence

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“…Under these circumstances, electrochemical technology has attracted attention as a detection method for PCR. − Electrochemistry is capable of detecting target DNA regardless of Debye length and can work with a wide range of ionic solutions. − …”

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confidence: 99%

Electrochemical Impedimetric Real-Time Polymerase Chain Reactions Using Anomalous Charge Transfer Enhancement

et al. 2022

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We developed a new electrochemical impedimetric method for the real-time detection of polymerase chain reactions (PCR) based on our recent discovery that the DNA intercalator, [Ru(bpy) 2 DPPZ] 2+ , anomalously enhances charge transfer between redox mediators, K 4 [Fe(CN) 6 ]/K 3 [Fe(CN) 6 ], and a carbon electrode. Three mM [Fe(CN) 6 ] 3−/4− and 5 μM [Ru-(bpy) 2 DPPZ] 2+ were added to the PCR solution, and electrochemical impedance spectroscopy (EIS) measurements were performed at each elongation heat cycle. The charge transfer resistance (Rct) was initially low due to the presence of [Ru(bpy) 2 DPPZ] 2+ in the solution. As PCR progressed, amplicon dsDNA was produced exponentially, and intercalated [Ru-(bpy) 2 DPPZ] 2+ ions, which could be detected as a steep Rct, increased at specific heat cycles depending on the amount of template DNA. The Rct increase per heat cycle, ΔRct, showed a peak at the same heat cycle as optical detection, proving that PCR can be accurately monitored in real time by impedance measurement. This simple method will enable a cost-effective and portable PCR device.

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“…There have been successful studies of electrochemical PCR detection in real time (qPCR), which used voltammetry measurement techniques to detect changes in redox peaks. − However, these published methods required specifically designed electric circuits.…”

mentioning

confidence: 99%

Anomalous Enhancement of Electrochemical Charge Transfer by a Ru Complex Ion Intercalator

2021

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We have found that the DNA intercalator [Ru(bpy) 2 DPPZ] 2+ (bpy = 2,2′-bipyridine; DPPZ = dipyrido[3,2-a:2′,3′c]phenazine) causes an anomalous increase in charge transfer in electrochemical impedance spectroscopy (EIS). With a carbonaceous electrode and a 1 mM hexacyanoferrate (1 mM [Fe(CN) 6 ] 3− and 1 mM [Fe(CN) 6 ] 4− ) mediator, we found that adding only 1 μM [Ru(bpy) 2 DPPZ] 2+ greatly enhanced the charge transfer between the electrode and hexacyanoferrate mediator, independently of other electrolytes or buffer components. The effect started with a one millionth amount of hexacyanoferrate. Since [Ru(bpy) 2 DPPZ] 2+ can intercalate with dsDNA, the effect is highly applicable for dsDNA detection or PCR monitoring. With further developments of this method, EIS sensors not requiring specific electrode modifications should be possible.

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Detection of a few DNA copies by real-time electrochemical polymerase chain reaction

Cited by 11 publications

References 31 publications

Computational Intelligence Methods for the Diagnosis of COVID-19

Computational Intelligence Methods for the Diagnosis of COVID-19

Electrochemical Impedimetric Real-Time Polymerase Chain Reactions Using Anomalous Charge Transfer Enhancement

Anomalous Enhancement of Electrochemical Charge Transfer by a Ru Complex Ion Intercalator

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