Smartphone-Based SARS-CoV-2 and Variants Detection System using Colorimetric DNAzyme Reaction Triggered by Loop-Mediated Isothermal Amplification (LAMP) with Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR)

Jayeon Song, Baekdong Cha, Jeong Moon, Hyowon Jang, Sunjoo Kim, Jieun Jang, Dongeun Yong, Hyung Jun Kwon, In Chul Lee, Eun Kyung Lim, Juyeon Jung, Hyun Gyu Park, Taejoon Kang

Research output: Contribution to journalArticlepeer-review

Abstract

Coronavirus disease (COVID-19) has affected people for over two years. Moreover, the emergence of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) variants has raised concerns regarding its accurate diagnosis. Here, we report a colorimetric DNAzyme reaction triggered by loop-mediated isothermal amplification (LAMP) with clustered regularly interspaced short palindromic repeats (CRISPR), referred to as DAMPR assay for detecting SARS-CoV-2 and variants genes with attomolar sensitivity within an hour. The CRISPR-associated protein 9 (Cas9) system eliminated false-positive signals of LAMP products, improving the accuracy of DAMPR assay. Further, we fabricated a portable DAMPR assay system using a three-dimensional printing technique and developed a machine learning (ML)-based smartphone application to routinely check diagnostic results of SARS-CoV-2 and variants. Among blind tests of 136 clinical samples, the proposed system successfully diagnosed COVID-19 patients with a clinical sensitivity and specificity of 100% each. More importantly, the D614G (variant-common), T478K (delta-specific), and A67V (omicron-specific) mutations of the SARS-CoV-2 S gene were detected selectively, enabling the diagnosis of 70 SARS-CoV-2 delta or omicron variant patients. The DAMPR assay system is expected to be employed for on-site, rapid, accurate detection of SARS-CoV-2 and its variants gene and employed in the diagnosis of various infectious diseases.

Original languageEnglish
Pages (from-to)11300-11314
Number of pages15
JournalACS Nano
Volume16
Issue number7
DOIs
Publication statusPublished - 2022 Jul 26

Bibliographical note

Funding Information:
This research was supported by National R&D Programs through National Research Foundation (NRF) of Korea funded by Ministry of Science and ICT (MSIT) of Korea (NRF-2021M3E5E3080379, NRF-2021M3E5E3080382, NRF-2021M3H4A1A02051048, NRF-2018M3A9E2022821, NRF-2021M3E5E3080844, NRF-2022R1C1C1008815, and NRF-2020R1A2C1010453), Global Frontier Program through Center for BioNano Health-Guard funded by MSIT of Korea (H-GUARD_2014M3A6B2060507 and H-GUARD_2013M3A6B2078950), R&D Program through National Research Council of Science & Technology (NST) of Korea funded by MSIT of Korea (No. CRC21021-100), Technology Development Program for Biological Hazards Management in Indoor Air through Korea Environment Industry & Technology Institute (KEITI) funded by Ministry of Environment (ME) of Korea (2021003370003), K-Sensor Technology Development Program funded by Ministry of Trade, Industry, and Energy (MOTIE) of Korea (RS-2022-00154855), Nanomedical Devices Development Program of National Nano Fab Center (CSM2105M101), and KRIBB Research Initiative Program (1711134081).

Funding Information:
This researchwas supported by National R&D Programs through National Research Foundation (NRF) of Korea funded by Ministry of Science and ICT (MSIT) of Korea (NRF-2021M3E5E3080379, NRF-2021M3E5E3080382NRF-2021M3H4A1A02051048, NRF-2018M3A9E2022821, NRF-2021M3E5E3080844NRF-2022R1C1C1008815, and NRF-2020R1A2C1010453), Global Frontier Programthrough Center for BioNano Health-Guard funded by MSIT of Korea (H-GUARD_2014M3A6B2060507and H-GUARD_2013M3A6B2078950), R&D Program through National Research Council of Science & Technology (NST) of Korea funded by MSITof Korea (No. CRC21021-100), Technology Development Program for BiologicalHazards Management in Indoor Air through Korea Environment Industry& Technology Institute (KEITI) funded by Ministry of Environment(ME) of Korea (2021003370003), K-Sensor Technology Development Programfunded by Ministry of Trade, Industry, and Energy (MOTIE) of Korea(RS-2022-00154855), Nanomedical Devices Development Program of NationalNano Fab Center (CSM2105M101), and KRIBB Research Initiative Program(1711134081).

Publisher Copyright:
© 2022 American Chemical Society.

All Science Journal Classification (ASJC) codes

  • Materials Science(all)
  • Engineering(all)
  • Physics and Astronomy(all)

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