Electrochemical genosensing of e. Coli based on padlock probes and rolling circle amplification

Alejandra Ben Aissa, Narayanan Madaboosi, Mats Nilsson, Maria Isabel Pividori*

*Corresponding author for this work

Research output: Contribution to journalArticleResearchpeer-review

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 en-demic, and the lack of reliable power supply, trained personnel and specialized facilities pose criti-cal 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 pre-concentrated 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 im-proved analytical performance, while simultaneously avoiding the use of thermocyclers or bulky bench top equipment.

Original languageEnglish
Article number5
Pages (from-to)1-13
Number of pages13
JournalSensors
Volume21
Issue number5
DOIs
Publication statusPublished - 3 Mar 2021

Keywords

  • Electrochemical genosensing
  • Isothermal amplification
  • Magnetic particles
  • Padlock probes
  • Rolling circle amplification

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