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Chembiochem . Amplification- and Enzyme-Free Magnetic Diagnostics Circuit for Whole-Genome Detection of SARS-CoV-2 RNA

tetano

Editor, Senior Moderator
Chembiochem


. 2024 May 6:e202400251.
doi: 10.1002/cbic.202400251. Online ahead of print. Amplification- and Enzyme-Free Magnetic Diagnostics Circuit for Whole-Genome Detection of SARS-CoV-2 RNA

Enja Laureen Rösch[SUP] 1 [/SUP], Rebecca Sack[SUP] 1 [/SUP], Mohammad Suman Chowdhury[SUP] 1 [/SUP], Florian Wolgast[SUP] 1 [/SUP], Margarete Zaborski[SUP] 2 [/SUP], Frank Ludwig[SUP] 1 [/SUP], Meinhard Schilling[SUP] 1 [/SUP], Thilo Viereck[SUP] 1 [/SUP], Ulfert Rand[SUP] 2 [/SUP], Aidin Lak[SUP] 3 [/SUP]



Affiliations
Abstract

Polymerase chain reaction (PCR) requires thermal cycling and enzymatic reactions for sequence amplification, hampering their applications in point-of-care (POC) settings. Magnetic bioassays based on magnetic particle spectroscopy (MPS) and magnetic nanoparticles (MNPs) are isothermal, wash-free, and can be quantitative. Realizing them amplification- and enzyme-free on a benchtop device, they will become irreplaceable for POC applications. Here we demonstrate a first-in-class magnetic signal amplification circuit (MAC) that enables detection of whole genome of SARS-CoV-2 by combining the specificity of toehold-mediated DNA strand displacement with the magnetic response of MNPs to declustering processes. Using MAC, we detect the N gene of SARS-CoV-2 samples at a concentration of 104 RNA copies/µl as determined by droplet digital PCR. Further, we demonstrate that MAC can reliably distinguish between SARS-CoV-2 and other human coronaviruses. Being a wash-, amplification- and enzyme-free biosensing concept and working at isothermal conditions (25 °C) on a low-cost benchtop MPS device, our MAC biosensing concept offers several indispensable features for translating nucleic acid detection to POC applications.

Keywords: Amplification-free assays; Magnetic particle spectroscopy; Magnetic-DNA nanoswitches; SARS-CoV-2 virus; Whole-genome detection.

 
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