Detection of Coronaviruses Using RNA Toehold Switch Sensors

• Published in: International journal of molecular sciences Vol. 22; no. 4; p. 1772
• Main Authors: Park, Soan, Lee, Jeong Wook
• Format: Journal Article
• Language: English
• Published: Switzerland MDPI 10.02.2021; MDPI AG
• Subjects: Coronavirus - genetics; Coronavirus - isolation & purification; Coronavirus Infections - diagnosis; Diagnostic Tests, Routine; Humans; middle east respiratory syndrome coronavirus (MERS-CoV); Molecular Diagnostic Techniques; molecular diagnostics; Nucleic Acid Amplification Techniques; Point-of-Care Testing; reverse transcription loop-mediated isothermal amplification (RT-LAMP); RNA, Viral - genetics; Sensitivity and Specificity; severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2); middle east respiratory syndrome coronavirus (MERS-CoV), severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), molecular diagnostics; reverse transcription loop-mediated isothermal amplification (RT-LAMP)
• ISSN: 1422-0067; 1661-6596; 1422-0067
• DOI: 10.3390/ijms22041772

A rapid, sensitive and simple point-of-care (POC) nucleic acid diagnostic test is needed to prevent spread of infectious diseases. Paper-based toehold reaction, a recently emerged colorimetric POC nucleic acid diagnostic test, has been widely used for pathogen detection and microbiome profiling. Here, we introduce an amplification method called reverse transcription loop-mediated amplification (RT-LAMP) prior to the toehold reaction and modify it to enable more sensitive and faster colorimetric detection of RNA viruses. We show that incorporating the modified RT-LAMP to the toehold reaction detects as few as 120 copies of coronavirus RNA in 70 min. Cross-reactivity test against other coronaviruses indicates this toehold reaction with the modified RT-LAMP is highly specific to the target RNA. Overall, the paper-based toehold switch sensors with the modified RT-LAMP allow fast, sensitive, specific and colorimetric coronavirus detection.