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A Novel Reverse Transcription Loop-Mediated Isothermal Amplification Method for Rapid Detection of SARS-CoV-2.
Int J Mol Sci. 2020 Apr 18; 21(8)IJ

Abstract

COVID-19 has become a major global public health burden, currently causing a rapidly growing number of infections and significant morbidity and mortality around the world. Early detection with fast and sensitive assays and timely intervention are crucial for interrupting the spread of the COVID-19 virus (SARS-CoV-2). Using a mismatch-tolerant amplification technique, we developed a simple, rapid, sensitive and visual reverse transcription loop-mediated isothermal amplification (RT-LAMP) assay for SARS-CoV-2 detection based on its N gene. The assay has a high specificity and sensitivity, and robust reproducibility, and its results can be monitored using a real-time PCR machine or visualized via colorimetric change from red to yellow. The limit of detection (LOD) of the assay is 118.6 copies of SARS-CoV-2 RNA per 25 μL reaction. The reaction can be completed within 30 min for real-time fluorescence monitoring, or 40 min for visual detection when the template input is more than 200 copies per 25 μL reaction. To evaluate the viability of the assay, a comparison between the RT-LAMP and a commercial RT-qPCR assay was made using 56 clinical samples. The SARS-CoV-2 RT-LAMP assay showed perfect agreement in detection with the RT-qPCR assay. The newly-developed SARS-CoV-2 RT-LAMP assay is a simple and rapid method for COVID-19 surveillance.

Authors+Show Affiliations

Clinical Laboratory, Nantong Third Hospital Affiliated to Nantong University, Nantong 226006, China.Pathogen Discovery and Evolution Unit, Institut Pasteur of Shanghai, Chinese Academy of Sciences, Shanghai 200031, China.Medical Laboratory of Taizhou Fourth People's Hospital, Taizhou 225300 China.Pathogen Discovery and Evolution Unit, Institut Pasteur of Shanghai, Chinese Academy of Sciences, Shanghai 200031, China.Shanghai Public Health Clinical Center, Fudan University, Jinshan District, Shanghai 201508, China.Pathogen Discovery and Evolution Unit, Institut Pasteur of Shanghai, Chinese Academy of Sciences, Shanghai 200031, China.

Pub Type(s)

Journal Article

Language

eng

PubMed ID

32325642

Citation

Lu, Renfei, et al. "A Novel Reverse Transcription Loop-Mediated Isothermal Amplification Method for Rapid Detection of SARS-CoV-2." International Journal of Molecular Sciences, vol. 21, no. 8, 2020.
Lu R, Wu X, Wan Z, et al. A Novel Reverse Transcription Loop-Mediated Isothermal Amplification Method for Rapid Detection of SARS-CoV-2. Int J Mol Sci. 2020;21(8).
Lu, R., Wu, X., Wan, Z., Li, Y., Jin, X., & Zhang, C. (2020). A Novel Reverse Transcription Loop-Mediated Isothermal Amplification Method for Rapid Detection of SARS-CoV-2. International Journal of Molecular Sciences, 21(8). https://doi.org/10.3390/ijms21082826
Lu R, et al. A Novel Reverse Transcription Loop-Mediated Isothermal Amplification Method for Rapid Detection of SARS-CoV-2. Int J Mol Sci. 2020 Apr 18;21(8) PubMed PMID: 32325642.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - A Novel Reverse Transcription Loop-Mediated Isothermal Amplification Method for Rapid Detection of SARS-CoV-2. AU - Lu,Renfei, AU - Wu,Xiuming, AU - Wan,Zhenzhou, AU - Li,Yingxue, AU - Jin,Xia, AU - Zhang,Chiyu, Y1 - 2020/04/18/ PY - 2020/03/19/received PY - 2020/04/04/revised PY - 2020/04/16/accepted PY - 2020/4/25/entrez PY - 2020/4/25/pubmed PY - 2020/4/28/medline KW - COVID-19 KW - LAMP KW - POCT KW - SARS-CoV-2 KW - pneumonia JF - International journal of molecular sciences JO - Int J Mol Sci VL - 21 IS - 8 N2 - COVID-19 has become a major global public health burden, currently causing a rapidly growing number of infections and significant morbidity and mortality around the world. Early detection with fast and sensitive assays and timely intervention are crucial for interrupting the spread of the COVID-19 virus (SARS-CoV-2). Using a mismatch-tolerant amplification technique, we developed a simple, rapid, sensitive and visual reverse transcription loop-mediated isothermal amplification (RT-LAMP) assay for SARS-CoV-2 detection based on its N gene. The assay has a high specificity and sensitivity, and robust reproducibility, and its results can be monitored using a real-time PCR machine or visualized via colorimetric change from red to yellow. The limit of detection (LOD) of the assay is 118.6 copies of SARS-CoV-2 RNA per 25 μL reaction. The reaction can be completed within 30 min for real-time fluorescence monitoring, or 40 min for visual detection when the template input is more than 200 copies per 25 μL reaction. To evaluate the viability of the assay, a comparison between the RT-LAMP and a commercial RT-qPCR assay was made using 56 clinical samples. The SARS-CoV-2 RT-LAMP assay showed perfect agreement in detection with the RT-qPCR assay. The newly-developed SARS-CoV-2 RT-LAMP assay is a simple and rapid method for COVID-19 surveillance. SN - 1422-0067 UR - https://www.unboundmedicine.com/medline/citation/32325642/A_Novel_Reverse_Transcription_Loop_Mediated_Isothermal_Amplification_Method_for_Rapid_Detection_of_SARS_CoV_2_ L2 - https://www.mdpi.com/resolver?pii=ijms21082826 DB - PRIME DP - Unbound Medicine ER -