Electrochemical Genosensing of E. coli Based on Padlock Probes and Rolling Circle Amplification
1
Grup de Sensors i Biosensors, Departament de Química, Universitat Autònoma de Barcelona, 08193 Bellaterra, Spain
2
IIT Madras Bioincubator, Indian Institute of Technology, Chennai 600113, India
3
Science for Life Laboratory, Department of Biochemistry and Biophysics, Stockholm University, 11419 Stockholm, Sweden
4
Institute of Biotechnology and Biomedicine, Universitat Autònoma de Barcelona, 08193 Bellaterra, Spain
*
Author to whom correspondence should be addressed.
Academic Editor: Giovanni Neri
Sensors 2021, 21(5), 1749; https://doi.org/10.3390/s21051749
Received: 30 December 2020
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Revised: 23 February 2021
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Accepted: 25 February 2021
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Published: 3 March 2021
(This article belongs to the Section Biosensors)
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.
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Keywords:
isothermal amplification; rolling circle amplification; padlock probes; electrochemical genosensing; magnetic particles
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MDPI and ACS Style
Ben Aissa, A.; Madaboosi, N.; Nilsson, M.; Pividori, M.I. Electrochemical Genosensing of E. coli Based on Padlock Probes and Rolling Circle Amplification. Sensors 2021, 21, 1749. https://doi.org/10.3390/s21051749
AMA Style
Ben Aissa A, Madaboosi N, Nilsson M, Pividori MI. Electrochemical Genosensing of E. coli Based on Padlock Probes and Rolling Circle Amplification. Sensors. 2021; 21(5):1749. https://doi.org/10.3390/s21051749
Chicago/Turabian StyleBen Aissa, Alejandra, Narayanan Madaboosi, Mats Nilsson, and Maria I. Pividori. 2021. "Electrochemical Genosensing of E. coli Based on Padlock Probes and Rolling Circle Amplification" Sensors 21, no. 5: 1749. https://doi.org/10.3390/s21051749
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