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RT-LAMP for rapid diagnosis of coronavirus SARS-CoV-2.
Microb Biotechnol. 2020 07; 13(4):950-961.MB

Abstract

The pandemic coronavirus SARS-CoV-2 in the world has caused a large infected population suffering from COVID-19. To curb the spreading of the virus, WHO urgently demanded an extension of screening and testing; thus, a rapid and simple diagnostic method is needed. We applied a reverse transcription-loop-mediated isothermal amplification (RT-LAMP) to achieve the detection of SARS-CoV-2 in 30 min. We designed four sets of LAMP primers (6 primers in each set), targeting the viral RNA of SARS-CoV-2 in the regions of orf1ab, S gene and N gene. A colorimetric change was used to report the results, which enables the outcome of viral RNA amplification to be read by the naked eye without the need of expensive or dedicated instrument. The sensitivity can be 80 copies of viral RNA per ml in a sample. We validated the RT-LAMP method in a hospital in China, employing 16 clinic samples with 8 positives and 8 negatives. The testing results are consistent with the conventional RT-qPCR. In addition, we also show that one-step process without RNA extraction is feasible to achieve RNA amplification directly from a sample. This rapid, simple and sensitive RT-LAMP method paves a way for a large screening at public domain and hospitals, particularly regional hospitals and medical centres in rural areas.

Authors+Show Affiliations

Oxford Suzhou Centre for Advanced Research (OSCAR), University of Oxford, Suzhou Industrial Park, Jiangsu, China. Department of Engineering Science, University of Oxford, Parks Road, OX1 3PJ, Oxford, UK.Department of Engineering Science, University of Oxford, Parks Road, OX1 3PJ, Oxford, UK.Department of Engineering Science, University of Oxford, Parks Road, OX1 3PJ, Oxford, UK. Institute of Biomedical Engineering, Department of Engineering Science, University of Oxford, Oxford, OX3 7DQ, UK.Medical Laboratory of Shenzhen Luohu People's Hospital, Shenzhen, 518001, China.Medical Laboratory of Shenzhen Luohu People's Hospital, Shenzhen, 518001, China.Department of Engineering Science, University of Oxford, Parks Road, OX1 3PJ, Oxford, UK. Institute of Biomedical Engineering, Department of Engineering Science, University of Oxford, Oxford, OX3 7DQ, UK.Oxford Suzhou Centre for Advanced Research (OSCAR), University of Oxford, Suzhou Industrial Park, Jiangsu, China.Oxford Suzhou Centre for Advanced Research (OSCAR), University of Oxford, Suzhou Industrial Park, Jiangsu, China.Oxford Suzhou Centre for Advanced Research (OSCAR), University of Oxford, Suzhou Industrial Park, Jiangsu, China.Oxford Suzhou Centre for Advanced Research (OSCAR), University of Oxford, Suzhou Industrial Park, Jiangsu, China.Oxford Suzhou Centre for Advanced Research (OSCAR), University of Oxford, Suzhou Industrial Park, Jiangsu, China.Medical Laboratory of Shenzhen Luohu People's Hospital, Shenzhen, 518001, China.Oxford Suzhou Centre for Advanced Research (OSCAR), University of Oxford, Suzhou Industrial Park, Jiangsu, China.Oxford Suzhou Centre for Advanced Research (OSCAR), University of Oxford, Suzhou Industrial Park, Jiangsu, China. Department of Engineering Science, University of Oxford, Parks Road, OX1 3PJ, Oxford, UK. Institute of Biomedical Engineering, Department of Engineering Science, University of Oxford, Oxford, OX3 7DQ, UK.

Pub Type(s)

Evaluation Study
Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

32333644

Citation

Huang, Wei E., et al. "RT-LAMP for Rapid Diagnosis of Coronavirus SARS-CoV-2." Microbial Biotechnology, vol. 13, no. 4, 2020, pp. 950-961.
Huang WE, Lim B, Hsu CC, et al. RT-LAMP for rapid diagnosis of coronavirus SARS-CoV-2. Microb Biotechnol. 2020;13(4):950-961.
Huang, W. E., Lim, B., Hsu, C. C., Xiong, D., Wu, W., Yu, Y., Jia, H., Wang, Y., Zeng, Y., Ji, M., Chang, H., Zhang, X., Wang, H., & Cui, Z. (2020). RT-LAMP for rapid diagnosis of coronavirus SARS-CoV-2. Microbial Biotechnology, 13(4), 950-961. https://doi.org/10.1111/1751-7915.13586
Huang WE, et al. RT-LAMP for Rapid Diagnosis of Coronavirus SARS-CoV-2. Microb Biotechnol. 2020;13(4):950-961. PubMed PMID: 32333644.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - RT-LAMP for rapid diagnosis of coronavirus SARS-CoV-2. AU - Huang,Wei E, AU - Lim,Boon, AU - Hsu,Chia-Chen, AU - Xiong,Dan, AU - Wu,Wei, AU - Yu,Yejiong, AU - Jia,Huidong, AU - Wang,Yun, AU - Zeng,Yida, AU - Ji,Mengmeng, AU - Chang,Hong, AU - Zhang,Xiuming, AU - Wang,Hui, AU - Cui,Zhanfeng, Y1 - 2020/04/25/ PY - 2020/03/27/received PY - 2020/04/13/revised PY - 2020/04/14/accepted PY - 2020/4/26/pubmed PY - 2020/6/12/medline PY - 2020/4/26/entrez SP - 950 EP - 961 JF - Microbial biotechnology JO - Microb Biotechnol VL - 13 IS - 4 N2 - The pandemic coronavirus SARS-CoV-2 in the world has caused a large infected population suffering from COVID-19. To curb the spreading of the virus, WHO urgently demanded an extension of screening and testing; thus, a rapid and simple diagnostic method is needed. We applied a reverse transcription-loop-mediated isothermal amplification (RT-LAMP) to achieve the detection of SARS-CoV-2 in 30 min. We designed four sets of LAMP primers (6 primers in each set), targeting the viral RNA of SARS-CoV-2 in the regions of orf1ab, S gene and N gene. A colorimetric change was used to report the results, which enables the outcome of viral RNA amplification to be read by the naked eye without the need of expensive or dedicated instrument. The sensitivity can be 80 copies of viral RNA per ml in a sample. We validated the RT-LAMP method in a hospital in China, employing 16 clinic samples with 8 positives and 8 negatives. The testing results are consistent with the conventional RT-qPCR. In addition, we also show that one-step process without RNA extraction is feasible to achieve RNA amplification directly from a sample. This rapid, simple and sensitive RT-LAMP method paves a way for a large screening at public domain and hospitals, particularly regional hospitals and medical centres in rural areas. SN - 1751-7915 UR - https://www.unboundmedicine.com/medline/citation/32333644/RT_LAMP_for_rapid_diagnosis_of_coronavirus_SARS_CoV_2_ L2 - https://doi.org/10.1111/1751-7915.13586 DB - PRIME DP - Unbound Medicine ER -