Detection of a few DNA copies by real-time electrochemical polymerase chain reaction - Sorbonne Université Access content directly
Journal Articles Analyst Year : 2017

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

Abstract

In the current work, accurate quantification over 10 to 10 8 DNA copies has been successfully achieved for the first time by real-time electrochemical PCR. This has been made possible thanks to the combined use of a fully automated house-built electrochemical qPCR device, optimized for parallel heating and electro-chemical monitoring of up to 48 PCR solutions, and the appropriate selection of a DNA intercalating redox probe retaining a strong affinity binding to ds-DNA at the PCR measurement temperature of 72 °C (corresponding to the PCR elongation step). This has also been achieved through the identification of the key parameters governing the onset electrochemical signal decrease and amplitude signal decrease as a function of the PCR cycle for a given DNA intercalating redox probe, thus allowing us to predict the electrochemical PCR kinetic plots from the values of the DNA affinity binding constant determined as a function of temperature. To the best of our knowledge, the analytical performances of the current electrochemical qPCR outperform all of those previously published, in terms of detection limit, dynamic range, reproducibility and melting curve analysis compared to that achieved on a commercialized bench-top fluorescence-based qPCR instrument.
Fichier principal
Vignette du fichier
Moreau2017.pdf (260.83 Ko) Télécharger le fichier
Origin Files produced by the author(s)

Dates and versions

hal-01929872 , version 1 (08-06-2022)

Licence

Identifiers

Cite

M. Moreau, S. Delile, A. Sharma, C. Fave, A. Perrier, et al.. Detection of a few DNA copies by real-time electrochemical polymerase chain reaction. Analyst, 2017, 142 (18), pp.3432-3440. ⟨10.1039/c7an00978j⟩. ⟨hal-01929872⟩
83 View
56 Download

Altmetric

Share

Gmail Mastodon Facebook X LinkedIn More