tetano
Editor, Senior Moderator
Exp Biol Med (Maywood)
. 2020 Nov 5;1535370220963793.
doi: 10.1177/1535370220963793. Online ahead of print.
Detection of severe acute respiratory syndrome coronavirus 2 and influenza viruses based on CRISPR-Cas12a
Oraphan Mayuramart[SUP] 1 [/SUP], Pattaraporn Nimsamer[SUP] 1 [/SUP], Somruthai Rattanaburi[SUP] 1 [/SUP], Naphat Chantaravisoot[SUP] 1 [/SUP], Kritsada Khongnomnan[SUP] 2 [/SUP], Jira Chansaenroj[SUP] 3 [/SUP], Jiratchaya Puenpa[SUP] 3 [/SUP], Nungruthai Suntronwong[SUP] 3 [/SUP], Preeyaporn Vichaiwattana[SUP] 3 [/SUP], Yong Poovorawan[SUP] 3 [/SUP], Sunchai Payungporn[SUP] 1 4 [/SUP]
Affiliations
Abstract
Due to the common symptoms of COVID-19, patients are similar to influenza-like illness. Therefore, the detection method would be crucial to discriminate between SARS-CoV-2 and influenza virus-infected patients. In this study, CRISPR-Cas12a-based detection was applied for detection of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), influenza A virus, and influenza B virus which would be a practical and attractive application for screening of patients with COVID-19 and influenza in areas with limited resources. The limit of detection for SARS-CoV-2, influenza A, and influenza B detection was 10, 10[SUP]3[/SUP], and 10[SUP]3[/SUP] copies/reaction, respectively. Moreover, the assays yielded no cross-reactivity against other respiratory viruses. The results revealed that the detection of influenza virus and SARS-CoV-2 by using RT-RPA and CRISPR-Cas12a technology reaches 96.23% sensitivity and 100% specificity for SARS-CoV-2 detection. The sensitivity for influenza virus A and B detections was 85.07% and 94.87%, respectively. In addition, the specificity for influenza virus A and B detections was approximately 96%. In conclusion, the RT-RPA with CRISPR-Cas12a assay was an effective method for the screening of influenza viruses and SARS-CoV-2 which could be applied to detect other infectious diseases in the future.
Keywords: CRISPR-Cas; Detection; RT-RPA; SARS-CoV-2; influenza A; influenza B.
. 2020 Nov 5;1535370220963793.
doi: 10.1177/1535370220963793. Online ahead of print.
Detection of severe acute respiratory syndrome coronavirus 2 and influenza viruses based on CRISPR-Cas12a
Oraphan Mayuramart[SUP] 1 [/SUP], Pattaraporn Nimsamer[SUP] 1 [/SUP], Somruthai Rattanaburi[SUP] 1 [/SUP], Naphat Chantaravisoot[SUP] 1 [/SUP], Kritsada Khongnomnan[SUP] 2 [/SUP], Jira Chansaenroj[SUP] 3 [/SUP], Jiratchaya Puenpa[SUP] 3 [/SUP], Nungruthai Suntronwong[SUP] 3 [/SUP], Preeyaporn Vichaiwattana[SUP] 3 [/SUP], Yong Poovorawan[SUP] 3 [/SUP], Sunchai Payungporn[SUP] 1 4 [/SUP]
Affiliations
- PMID: 33153299
- DOI: 10.1177/1535370220963793
Abstract
Due to the common symptoms of COVID-19, patients are similar to influenza-like illness. Therefore, the detection method would be crucial to discriminate between SARS-CoV-2 and influenza virus-infected patients. In this study, CRISPR-Cas12a-based detection was applied for detection of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), influenza A virus, and influenza B virus which would be a practical and attractive application for screening of patients with COVID-19 and influenza in areas with limited resources. The limit of detection for SARS-CoV-2, influenza A, and influenza B detection was 10, 10[SUP]3[/SUP], and 10[SUP]3[/SUP] copies/reaction, respectively. Moreover, the assays yielded no cross-reactivity against other respiratory viruses. The results revealed that the detection of influenza virus and SARS-CoV-2 by using RT-RPA and CRISPR-Cas12a technology reaches 96.23% sensitivity and 100% specificity for SARS-CoV-2 detection. The sensitivity for influenza virus A and B detections was 85.07% and 94.87%, respectively. In addition, the specificity for influenza virus A and B detections was approximately 96%. In conclusion, the RT-RPA with CRISPR-Cas12a assay was an effective method for the screening of influenza viruses and SARS-CoV-2 which could be applied to detect other infectious diseases in the future.
Keywords: CRISPR-Cas; Detection; RT-RPA; SARS-CoV-2; influenza A; influenza B.