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

Carinelli S, K├╝hnemund M, Nilsson M, Pividori MI

Biosens Bioelectron 93 (-) 65-71 [2017-07-15; online 2016-10-11]

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.

Affiliated researcher

PubMed 27838201

DOI 10.1016/j.bios.2016.09.099

Crossref 10.1016/j.bios.2016.09.099

pii: S0956-5663(16)30983-6

Publications 9.5.0