tetano
Editor, Senior Moderator
J Clin Virol
. 2020 May 21;129:104446.
doi: 10.1016/j.jcv.2020.104446. Online ahead of print.
Evaluation of Rapid Diagnosis of Novel Coronavirus Disease (COVID-19) Using Loop-Mediated Isothermal Amplification
Yutaro Kitagawa[SUP] 1 [/SUP], Yuta Orihara[SUP] 1 [/SUP], Rieko Kawamura[SUP] 1 [/SUP], Kazuo Imai[SUP] 2 [/SUP], Jun Sakai[SUP] 2 [/SUP], Norihito Tarumoto[SUP] 2 [/SUP], Masaru Matsuoka[SUP] 1 [/SUP], Shinichi Takeuchi[SUP] 1 [/SUP], Shigefumi Maesaki[SUP] 2 [/SUP], Takuya Maeda[SUP] 3 [/SUP]
Affiliations
Abstract
With the rapid spread of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), there is an urgent need for more rapid and simple detection technologies at the forefront of medical care worldwide. In this study, we evaluated the effectiveness of the Loopamp? 2019-SARSCoV-2 Detection Reagent Kit, which uses loop-mediated isothermal amplification (LAMP) technology. In this protocol, cDNA is synthesized from SARS-CoV-2 RNA using reverse transcriptase, followed by DNA amplification under isothermal conditions in one step. The RT-LAMP test kit amplified the targeted RNA of a SARS-CoV-2 isolate with a detection limit of 1.0 ? 101 copies/μL, which was comparable to the detection sensitivity of quantitative reverse transcription PCR (RT-qPCR). Comparison with the results of RT-qPCR for 76 nasopharyngeal swab samples from patients with suspected COVID-19 showed a sensitivity of 100 % and a specificity of 97.6 %. In the 24 RNA specimens derived from febrile Japanese patients with or without influenza A, no amplification was observed using RT-LAMP. RT-LAMP could be a simple and easy-to-use diagnostic tool for the detection of SARS-CoV-2.
Keywords: COVID-19; LAMP; RT-qPCR; SARS-CoV-2.
. 2020 May 21;129:104446.
doi: 10.1016/j.jcv.2020.104446. Online ahead of print.
Evaluation of Rapid Diagnosis of Novel Coronavirus Disease (COVID-19) Using Loop-Mediated Isothermal Amplification
Yutaro Kitagawa[SUP] 1 [/SUP], Yuta Orihara[SUP] 1 [/SUP], Rieko Kawamura[SUP] 1 [/SUP], Kazuo Imai[SUP] 2 [/SUP], Jun Sakai[SUP] 2 [/SUP], Norihito Tarumoto[SUP] 2 [/SUP], Masaru Matsuoka[SUP] 1 [/SUP], Shinichi Takeuchi[SUP] 1 [/SUP], Shigefumi Maesaki[SUP] 2 [/SUP], Takuya Maeda[SUP] 3 [/SUP]
Affiliations
- PMID: 32512376
- PMCID: PMC7241399
- DOI: 10.1016/j.jcv.2020.104446
Abstract
With the rapid spread of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), there is an urgent need for more rapid and simple detection technologies at the forefront of medical care worldwide. In this study, we evaluated the effectiveness of the Loopamp? 2019-SARSCoV-2 Detection Reagent Kit, which uses loop-mediated isothermal amplification (LAMP) technology. In this protocol, cDNA is synthesized from SARS-CoV-2 RNA using reverse transcriptase, followed by DNA amplification under isothermal conditions in one step. The RT-LAMP test kit amplified the targeted RNA of a SARS-CoV-2 isolate with a detection limit of 1.0 ? 101 copies/μL, which was comparable to the detection sensitivity of quantitative reverse transcription PCR (RT-qPCR). Comparison with the results of RT-qPCR for 76 nasopharyngeal swab samples from patients with suspected COVID-19 showed a sensitivity of 100 % and a specificity of 97.6 %. In the 24 RNA specimens derived from febrile Japanese patients with or without influenza A, no amplification was observed using RT-LAMP. RT-LAMP could be a simple and easy-to-use diagnostic tool for the detection of SARS-CoV-2.
Keywords: COVID-19; LAMP; RT-qPCR; SARS-CoV-2.