Electrical Detection Assay Based on Programmable Nucleic Acid Probe for Efficient Single-Nucleotide Polymorphism Identification
- 17 May 2023
- journal article
- research article
- Published by American Chemical Society (ACS) in ACS Sensors
- Vol. 8 (5), 2096-2104
- https://doi.org/10.1021/acssensors.3c00453
Abstract
Thelarge-scale pandemic and fast evolution of severe acute respiratorysyndrome coronavirus 2 (SARS-CoV-2) variants have triggered an urgentneed for an efficient and sensitive on-site nucleic acid testing methodwith single-nucleotide polymorphism (SNP) identification capability.Here, we report a multiplexed electrical detection assay based ona paperclip-shaped nucleic acid probe (PNprobe) functionalized field-effecttransistor (FET) biosensor for highly sensitive and specific detectionand discrimination of SARS-CoV-2 variants. The three-stem structureof the PNprobe significantly amplifies the thermodynamic stabilitydifference between variant RNAs that differ in a single-nucleotidemutation. With the assistance of combinatorial FET detection channels,the assay realizes simultaneously the detection and identificationof key mutations of seven SARS-CoV-2 variants, including nucleotidesubstitutions and deletions at single-nucleotide resolution within15 min. For 70 simulated throat swab samples, the multiplexed electricaldetection assay shows an identification accuracy of 97.1% for thediscrimination of SARS-CoV-2 variants. Our designed multiplexed electricaldetection assay with SNP identification capability provides an efficienttool to achieve scalable pandemic screening.Keywords
Funding Information
- National Key Research and Development Program of China (2021YFA1202400)
- Fundamental Research Funds for the Central Universities (2042022rc0004)
- National Natural Science Foundation of China (21925401)
- Tencent Foundation
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