TY - JOUR T1 - Rapid and visual detection of 2019 novel coronavirus (SARS-CoV-2) by a reverse transcription loop-mediated isothermal amplification assay. AU - Yan,C, AU - Cui,J, AU - Huang,L, AU - Du,B, AU - Chen,L, AU - Xue,G, AU - Li,S, AU - Zhang,W, AU - Zhao,L, AU - Sun,Y, AU - Yao,H, AU - Li,N, AU - Zhao,H, AU - Feng,Y, AU - Liu,S, AU - Zhang,Q, AU - Liu,D, AU - Yuan,J, Y1 - 2020/04/08/ PY - 2020/03/03/received PY - 2020/04/01/revised PY - 2020/04/02/accepted PY - 2020/4/11/pubmed PY - 2020/6/2/medline PY - 2020/4/11/entrez KW - COVID-19 KW - Detection KW - RT-LAMP KW - SARS-CoV-2 KW - Visual SP - 773 EP - 779 JF - Clinical microbiology and infection : the official publication of the European Society of Clinical Microbiology and Infectious Diseases JO - Clin Microbiol Infect VL - 26 IS - 6 N2 - OBJECTIVE: To evaluate a reverse transcription loop-mediated isothermal amplification (RT-LAMP) assay for detection of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), and compare it with RT-PCR. METHODS: We designed primers specific to the orf1ab and S genes of SARS-CoV-2. Total viral RNA was extracted using the QIAamp Viral RNA Mini Kit. We optimized the RT-LAMP assay, and evaluated it for its sensitivity and specificity of detection using real-time turbidity monitoring and visual observation. RESULTS: The primer sets orf1ab-4 and S-123 amplified the genes in the shortest times, the mean (±SD) times were 18 ± 1.32 min and 20 ± 1.80 min, respectively, and 63°C was the optimum reaction temperature. The sensitivities were 2 × 101 copies and 2 × 102 copies per reaction with primer sets orf1ab-4 and S-123, respectively. This assay showed no cross-reactivity with 60 other respiratory pathogens. To describe the availability of this method in clinical diagnosis, we collected 130 specimens from patients with clinically suspected SARS-CoV-2 infection. Among them, 58 were confirmed to be positive and 72 were negative by RT-LAMP. The sensitivity was 100% (95% CI 92.3%-100%), specificity 100% (95% CI 93.7%-100%). This assay detected SARS-CoV-2 in a mean (±SD) time of 26.28 ± 4.48 min and the results can be identified with visual observation. CONCLUSION: These results demonstrate that we developed a rapid, simple, specific and sensitive RT-LAMP assay for SARS-CoV-2 detection among clinical samples. It will be a powerful tool for SARS-CoV-2 identification, and for monitoring suspected patients, close contacts and high-risk groups. SN - 1469-0691 UR - https://www.unboundmedicine.com/medline/citation/32276116/Rapid_and_visual_detection_of_2019_novel_coronavirus__SARS_CoV_2__by_a_reverse_transcription_loop_mediated_isothermal_amplification_assay_ L2 - https://linkinghub.elsevier.com/retrieve/pii/S1198-743X(20)30186-5 DB - PRIME DP - Unbound Medicine ER -