Colorimetric detection of SARS-CoV-2 and drug-resistant pH1N1 using CRISPR/dCas9

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Title
Colorimetric detection of SARS-CoV-2 and drug-resistant pH1N1 using CRISPR/dCas9
Author(s)
Jeong Moon; Hyung-Jun Kwon; D Yong; In Chul Lee; Hongki Kim; Hyunju Kang; Eun Kyung LimKyu-Sun LeeJuyeon Jung; H G Park; Taejoon Kang
Bibliographic Citation
ACS Sensors, vol. 5, pp. 4017-4026
Publication Year
2020
Abstract
Viruses have been a continuous threat to human beings. The coronavirus disease 2019 (COVID-19), caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), has led to a pandemic that is still ongoing worldwide. Previous pandemic influenza A virus (pH1N1) might be re-emerging through a drug-resistant mutation. We report a colorimetric viral detection method based on the clustered regularly interspaced short palindromic repeats (CRISPR)/Cas9 endonuclease dead (dCas9) system. In this method, RNA in the viral lysate was directly recognized by the CRISPR/dCas9 system with biotin-protospacer adjacent motif (PAM)-presenting oligonucleotide (PAMmer). Streptavidin-horseradish peroxidase then bound to biotin-PAMmer, inducing a color change through the oxidation of 3,3′,5,5′-tetramethylbenzidine. Using the developed method, we successfully identified SARS-CoV-2, pH1N1, and pH1N1/H275Y viruses by the naked eye. Moreover, the detection of viruses in human nasopharyngeal aspirates and sputum was demonstrated. Finally, clinical samples from COVID-19 patients led to a successful diagnosis. We anticipate that the current method can be employed for simple and accurate diagnosis of viruses.
Keyword
SARS-CoV-2COVID-19Drug-resistanceInfluenza virusCRISPR/dCas9Colorimetry
ISSN
2379-3694
Publisher
Amer Chem Soc
DOI
http://dx.doi.org/10.1021/acssensors.0c01929
Type
Article
Appears in Collections:
Jeonbuk Branch Institute > Functional Biomaterial Research Center > 1. Journal Articles
Division of Biomaterials Research > Bionanotechnology Research Center > 1. Journal Articles
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