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Development of a reverse transcription-loop-mediated isothermal amplification as a rapid early-detection method for novel SARS-CoV-2.
Emerg Microbes Infect. 2020 Dec; 9(1):998-1007.EM

Abstract

The previous outbreaks of SARS-CoV and MERS-CoV have led researchers to study the role of diagnostics in impediment of further spread and transmission. With the recent emergence of the novel SARS-CoV-2, the availability of rapid, sensitive, and reliable diagnostic methods is essential for disease control. Hence, we have developed a reverse transcription loop-mediated isothermal amplification (RT-LAMP) assay for the specific detection of SARS-CoV-2. The primer sets for RT-LAMP assay were designed to target the nucleocapsid gene of the viral RNA, and displayed a detection limit of 102 RNA copies close to that of qRT-PCR. Notably, the assay has exhibited a rapid detection span of 30 min combined with the colorimetric visualization. This test can detect specifically viral RNAs of the SARS-CoV-2 with no cross-reactivity to related coronaviruses, such as HCoV-229E, HCoV-NL63, HCoV-OC43, and MERS-CoV as well as human infectious influenza viruses (type B, H1N1pdm, H3N2, H5N1, H5N6, H5N8, and H7N9), and other respiratory disease-causing viruses (RSVA, RSVB, ADV, PIV, MPV, and HRV). Furthermore, the developed RT-LAMP assay has been evaluated using specimens collected from COVID-19 patients that exhibited high agreement to the qRT-PCR. Our RT-LAMP assay is simple to perform, less expensive, time-efficient, and can be used in clinical laboratories for preliminary detection of SARS-CoV-2 in suspected patients. In addition to the high sensitivity and specificity, this isothermal amplification conjugated with a single-tube colorimetric detection method may contribute to the public health responses and disease control, especially in the areas with limited laboratory capacities.

Authors+Show Affiliations

Department of Microbiology, Chungbuk National University College of Medicine and Medical Research Institute, Cheongju, Republic of Korea.Research Institute of Public Health, National Medical Center, Seoul, Republic of Korea.Department of Microbiology, Chungbuk National University College of Medicine and Medical Research Institute, Cheongju, Republic of Korea.Department of Microbiology, Chungbuk National University College of Medicine and Medical Research Institute, Cheongju, Republic of Korea.Center for Infectious Diseases Research, Department of Internal Medicine, National Medical Center, Seoul, Republic of Korea.Department of Microbiology, Chungbuk National University College of Medicine and Medical Research Institute, Cheongju, Republic of Korea.Department of Microbiology, Chungbuk National University College of Medicine and Medical Research Institute, Cheongju, Republic of Korea.Department of Microbiology, Chungbuk National University College of Medicine and Medical Research Institute, Cheongju, Republic of Korea.Department of Microbiology, Chungbuk National University College of Medicine and Medical Research Institute, Cheongju, Republic of Korea.Department of Microbiology, Chungbuk National University College of Medicine and Medical Research Institute, Cheongju, Republic of Korea.Center for Infectious Diseases Research, Department of Internal Medicine, National Medical Center, Seoul, Republic of Korea.Department of Microbiology, Chungbuk National University College of Medicine and Medical Research Institute, Cheongju, Republic of Korea.School of Biological Sciences, Chungbuk National University, Cheongju, Republic of Korea.Department of Laboratory Medicine, Chungbuk National University College of Medicine, Cheongju, Republic of Korea.Department of Microbiology, Chungbuk National University College of Medicine and Medical Research Institute, Cheongju, Republic of Korea.Research Institute of Public Health, National Medical Center, Seoul, Republic of Korea.Department of Microbiology, Chungbuk National University College of Medicine and Medical Research Institute, Cheongju, Republic of Korea.

Pub Type(s)

Journal Article

Language

eng

PubMed ID

32306853

Citation

Baek, Yun Hee, et al. "Development of a Reverse Transcription-loop-mediated Isothermal Amplification as a Rapid Early-detection Method for Novel SARS-CoV-2." Emerging Microbes & Infections, vol. 9, no. 1, 2020, pp. 998-1007.
Baek YH, Um J, Antigua KJC, et al. Development of a reverse transcription-loop-mediated isothermal amplification as a rapid early-detection method for novel SARS-CoV-2. Emerg Microbes Infect. 2020;9(1):998-1007.
Baek, Y. H., Um, J., Antigua, K. J. C., Park, J. H., Kim, Y., Oh, S., Kim, Y. I., Choi, W. S., Kim, S. G., Jeong, J. H., Chin, B. S., Nicolas, H. D. G., Ahn, J. Y., Shin, K. S., Choi, Y. K., Park, J. S., & Song, M. S. (2020). Development of a reverse transcription-loop-mediated isothermal amplification as a rapid early-detection method for novel SARS-CoV-2. Emerging Microbes & Infections, 9(1), 998-1007. https://doi.org/10.1080/22221751.2020.1756698
Baek YH, et al. Development of a Reverse Transcription-loop-mediated Isothermal Amplification as a Rapid Early-detection Method for Novel SARS-CoV-2. Emerg Microbes Infect. 2020;9(1):998-1007. PubMed PMID: 32306853.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Development of a reverse transcription-loop-mediated isothermal amplification as a rapid early-detection method for novel SARS-CoV-2. AU - Baek,Yun Hee, AU - Um,Jihye, AU - Antigua,Khristine Joy C, AU - Park,Ji-Hyun, AU - Kim,Yeonjae, AU - Oh,Sol, AU - Kim,Young-Il, AU - Choi,Won-Suk, AU - Kim,Seong Gyu, AU - Jeong,Ju Hwan, AU - Chin,Bum Sik, AU - Nicolas,Halcyon Dawn G, AU - Ahn,Ji-Young, AU - Shin,Kyeong Seob, AU - Choi,Young Ki, AU - Park,Jun-Sun, AU - Song,Min-Suk, PY - 2020/4/21/pubmed PY - 2020/5/20/medline PY - 2020/4/21/entrez KW - COVID-19 KW - SARS-CoV-2 KW - colorimetric detection KW - molecular diagnosis KW - novel coronavirus KW - reverse transcription-loop-mediated isothermal amplification SP - 998 EP - 1007 JF - Emerging microbes & infections JO - Emerg Microbes Infect VL - 9 IS - 1 N2 - The previous outbreaks of SARS-CoV and MERS-CoV have led researchers to study the role of diagnostics in impediment of further spread and transmission. With the recent emergence of the novel SARS-CoV-2, the availability of rapid, sensitive, and reliable diagnostic methods is essential for disease control. Hence, we have developed a reverse transcription loop-mediated isothermal amplification (RT-LAMP) assay for the specific detection of SARS-CoV-2. The primer sets for RT-LAMP assay were designed to target the nucleocapsid gene of the viral RNA, and displayed a detection limit of 102 RNA copies close to that of qRT-PCR. Notably, the assay has exhibited a rapid detection span of 30 min combined with the colorimetric visualization. This test can detect specifically viral RNAs of the SARS-CoV-2 with no cross-reactivity to related coronaviruses, such as HCoV-229E, HCoV-NL63, HCoV-OC43, and MERS-CoV as well as human infectious influenza viruses (type B, H1N1pdm, H3N2, H5N1, H5N6, H5N8, and H7N9), and other respiratory disease-causing viruses (RSVA, RSVB, ADV, PIV, MPV, and HRV). Furthermore, the developed RT-LAMP assay has been evaluated using specimens collected from COVID-19 patients that exhibited high agreement to the qRT-PCR. Our RT-LAMP assay is simple to perform, less expensive, time-efficient, and can be used in clinical laboratories for preliminary detection of SARS-CoV-2 in suspected patients. In addition to the high sensitivity and specificity, this isothermal amplification conjugated with a single-tube colorimetric detection method may contribute to the public health responses and disease control, especially in the areas with limited laboratory capacities. SN - 2222-1751 UR - https://www.unboundmedicine.com/medline/citation/32306853/Development_of_a_reverse_transcription_loop_mediated_isothermal_amplification_as_a_rapid_early_detection_method_for_novel_SARS_CoV_2_ L2 - https://www.tandfonline.com/doi/full/10.1080/22221751.2020.1756698 DB - PRIME DP - Unbound Medicine ER -