Electrochemical Genosensing of E. coli Based on Padlock Probes and Rolling Circle Amplification
Ben Aissa Soler, Alejandra (Universitat Autònoma de Barcelona. Departament de Química)
Madaboosi, Narayanan (Stockholm University)
Nilsson, Mats (Stockholm University)
Pividori, María Isabel (Universitat Autònoma de Barcelona. Departament de Química)

Date:	2021
Abstract:	Isothermal amplification techniques are emerging nowadays for the rapid and accurate detection of pathogenic bacteria in low resource settings, where many infectious diseases are endemic, and the lack of reliable power supply, trained personnel and specialized facilities pose critical barriers for timely diagnosis. This work addresses the detection of E. coli based on DNA isothermal amplification performed on magnetic particles (MPs) followed by electrochemical genosensing on disposable electrodes by square-wave voltammetry. In this approach, the bacterial DNA is preconcentrated using a target-specific magnetic probe and then amplified on the MPs by rolling circle amplification (RCA). Two different electrochemical readout methods for the RCA amplicons are tested. The first one relied on the labelling of the magnetic RCA product with a digoxigenin probe followed by the incubation with antiDIG-HRP antibody as electrochemical reporter. In the second case, the direct detection with an HRP-probe was performed. This latter strategy showed an improved analytical performance, while simultaneously avoiding the use of thermocyclers or bulky bench top equipment.
Grants:	Agencia Estatal de Investigación BIO2016-75751-R Agencia Estatal de Investigación PID2019-106625RB-I00
Rights:	Aquest document està subjecte a una llicència d'ús Creative Commons. Es permet la reproducció total o parcial, la distribució, la comunicació pública de l'obra i la creació d'obres derivades, fins i tot amb finalitats comercials, sempre i quan es reconegui l'autoria de l'obra original.
Language:	Anglès
Document:	Article ; recerca ; Versió publicada
Subject:	Isothermal amplification ; Rolling circle amplification ; Padlock probes ; Electrochemical genosensing ; Magnetic particles
Published in:	Sensors (Basel, Switzerland), Vol. 21, Issue 5 (March 2021) , art. 1749, ISSN 1424-8220

DOI: 10.3390/s21051749
PMID: 33802488

13 p, 2.4 MB

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