Yoctomole electrochemical genosensing of Ebola virus cDNA by rolling circle and circle to circle amplification

S. Carinelli, M. Kühnemund, M. Nilsson, M. I. Pividori

Research output: Contribution to journalArticleResearchpeer-review

18 Citations (Scopus)

Abstract

© 2016 Elsevier B.V. This work addresses the design of an Ebola diagnostic test involving a simple, rapid, specific and highly sensitive procedure based on isothermal amplification on magnetic particles with electrochemical readout. Ebola padlock probes were designed to detect a specific L-gene sequence present in the five most common Ebola species. Ebola cDNA was amplified by rolling circle amplification (RCA) on magnetic particles. Further re-amplification was performed by circle-to-circle amplification (C2CA) and the products were detected in a double-tagging approach using a biotinylated capture probe for immobilization on magnetic particles and a readout probe for electrochemical detection by square-wave voltammetry on commercial screen-printed electrodes. The electrochemical genosensor was able to detect as low as 200 ymol, corresponding to 120 cDNA molecules of L-gene Ebola virus with a limit of detection of 33 cDNA molecules. The isothermal double-amplification procedure by C2CA combined with the electrochemical readout and the magnetic actuation enables the high sensitivity, resulting in a rapid, inexpensive, robust and user-friendly sensing strategy that offers a promising approach for the primary care in low resource settings, especially in less developed countries.
Original languageEnglish
Pages (from-to)65-71
JournalBiosensors and Bioelectronics
Volume93
DOIs
Publication statusPublished - 15 Jul 2017

Keywords

  • Circle-to-circle amplification
  • Ebola virus
  • Electrochemical genosensing
  • Isothermal amplification
  • Magnetic particle

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