Tags

Type your tag names separated by a space and hit enter

Rapid and visual detection of 2019 novel coronavirus (SARS-CoV-2) by a reverse transcription loop-mediated isothermal amplification assay.
Clin Microbiol Infect. 2020 Jun; 26(6):773-779.CM

Abstract

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.

Authors+Show Affiliations

Capital Institute of Paediatrics, Beijing, China.Capital Institute of Paediatrics, Beijing, China.Treatment and Research Centre for Infectious Diseases, The Fifth Medical Centre of PLA General Hospital, Beijing, China.Capital Institute of Paediatrics, Beijing, China.Beijing Macro & Micro-test Bio-Tech Co., Ltd. Beijing, China.Capital Institute of Paediatrics, Beijing, China.Capital Institute of Paediatrics, Beijing, China.Capital Institute of Paediatrics, Beijing, China.Capital Institute of Paediatrics, Beijing, China.Capital Institute of Paediatrics, Beijing, China.Capital Institute of Paediatrics, Beijing, China.Capital Institute of Paediatrics, Beijing, China.Capital Institute of Paediatrics, Beijing, China.Capital Institute of Paediatrics, Beijing, China.Capital Institute of Paediatrics, Beijing, China.Capital Institute of Paediatrics, Beijing, China.Computational Virology Group, Centre for Bacteria and Virus Resources and Bioinformation, Wuhan Institute of Virology, Chinese Academy of Sciences, Wuhan, China. Electronic address: liud@wh.iov.cn.Capital Institute of Paediatrics, Beijing, China. Electronic address: yuanjing6216@163.com.

Pub Type(s)

Comparative Study
Journal Article

Language

eng

PubMed ID

32276116

Citation

Yan, C, et al. "Rapid and Visual Detection of 2019 Novel Coronavirus (SARS-CoV-2) By a Reverse Transcription Loop-mediated Isothermal Amplification Assay." Clinical Microbiology and Infection : the Official Publication of the European Society of Clinical Microbiology and Infectious Diseases, vol. 26, no. 6, 2020, pp. 773-779.
Yan C, Cui J, Huang L, et al. Rapid and visual detection of 2019 novel coronavirus (SARS-CoV-2) by a reverse transcription loop-mediated isothermal amplification assay. Clin Microbiol Infect. 2020;26(6):773-779.
Yan, C., Cui, J., Huang, L., Du, B., Chen, L., Xue, G., Li, S., Zhang, W., Zhao, L., Sun, Y., Yao, H., Li, N., Zhao, H., Feng, Y., Liu, S., Zhang, Q., Liu, D., & Yuan, J. (2020). Rapid and visual detection of 2019 novel coronavirus (SARS-CoV-2) by a reverse transcription loop-mediated isothermal amplification assay. Clinical Microbiology and Infection : the Official Publication of the European Society of Clinical Microbiology and Infectious Diseases, 26(6), 773-779. https://doi.org/10.1016/j.cmi.2020.04.001
Yan C, et al. Rapid and Visual Detection of 2019 Novel Coronavirus (SARS-CoV-2) By a Reverse Transcription Loop-mediated Isothermal Amplification Assay. Clin Microbiol Infect. 2020;26(6):773-779. PubMed PMID: 32276116.
* Article titles in AMA citation format should be in sentence-case
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 -