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.
Bibliographical noteFunding Information:
This research was supported by the Basic Science Research Program through the National Research Foundation of Korea (NRF) funded by the Ministry of Science and ICT (MSIT) (NRF-2019R1C1C1006867 and NRF-2020R1A2C1010453), the Center for BioNano HealthGuard funded by the MSIT of Korea as a Global Frontier Project (H-GUARD_2013M3A6B2078950 and H-GUARD_2014M3A6B2060507), the Bio & Medical Technology Development Program of the NRF funded by the MSIT of Korea (NRF-2018M3A9E2022821), KRIBB Research Initiative Program, and Kyung Nam Pharm. and Kyung Nam Biopharma. J.M. is a recipient of the Global Ph.D. Fellowship (NRF-2019H1A2A1073468).
All Science Journal Classification (ASJC) codes
- Process Chemistry and Technology
- Fluid Flow and Transfer Processes