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

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...

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Published in:Chembiochem : a European journal of chemical biology 2024-08, Vol.25 (16), p.e202400251
Main Authors: Rösch, Enja Laureen, Sack, Rebecca, Chowdhury, Mohammad Suman, Wolgast, Florian, Zaborski, Margarete, Ludwig, Frank, Schilling, Meinhard, Viereck, Thilo, Rand, Ulfert, Lak, Aidin
Format: Article
Language:English
Subjects:
Amplification
Bioassays
Biosensors
Coronaviruses
COVID-19
DNA-directed RNA polymerase
Enzymes
Genomes
Magnetic signals
N gene
Nanoparticles
Nucleic acids
Nucleotide sequence
Polymerase chain reaction
Ribonucleic acid
RNA
RNA polymerase
Severe acute respiratory syndrome
Severe acute respiratory syndrome coronavirus 2
Spectroscopy
Thermal cycling
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Summary: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 10 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.
ISSN:1439-4227
1439-7633
1439-7633
DOI:10.1002/cbic.202400251