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한국생명공학연구원

ACS Nano, Publication Date:June 23, 2022 | https://doi.org/10.1021/acsnano.2c04840 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,^# and Taejoon Kang^†,*

^†Bionanotechnology Research Center, Korea Research Institute of Bioscience and Biotechnology (KRIBB), 125 Gwahak-ro, Yuseong-gu, Daejeon 34141, Republic of Korea
^‡School of Integrated Technology, Gwangju Institute of Science and Technology (GIST), 123 Cheomdangwagiro, Buk-gu, Gwangju 61005, Republic of Korea
^#Department of Chemical and Biomolecular Engineering, Korea Advanced Institute of Science and Technology (KAIST), 291 Daehak-ro, Yuseong-gu, Daejeon 34141, Republic of Korea
^&Department of Laboratory Medicine, Gyeongsang National University College of Medicine, 79 Gangnam-ro, Jinju-si, Gyeongsangnam-do 52727, Republic of Korea
^||Gyeongnam Center for Disease Control and Prevention, 300 Jungang-daero, Uichang-gu, Changwon-si, Gyeongsangnamdo 51154, Republic of Korea
^§Department of Laboratory Medicine and Research Institute of Bacterial Resistance, Yonsei University College of Medicine, 50-1 Yonsei-ro, Seodaemun-gu, Seoul 03722, Republic of Korea
⁺Functional Biomaterial Research Center, KRIBB, 181 Ipsin-gil, Jeongeup-si, Jeollabuk-do 56212, Republic of Korea
^※Department of Nanobiotechnology, KRIBB School of Biotechnology, University of Science and Technology (UST), 217 Gajeong-ro, Yuseong-gu, Daejeon 34113, Republic of Korea

^¶These authors contributed equally to this work.
^*Corresponding author.

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.

논문정보

형식Research article
게재일2022년 06월 (BRIC 등록일 2022-08-05)
연구진국내 연구진
분야 바이오·의료융합 > 바이오센싱 및 나노바이오물질

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