Rapid point-of-care detection of SARS-CoV-2 using reverse transcription loop-mediated isothermal amplification (RT-LAMP)
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METHODOLOGY
Rapid point‑of‑care detection of SARS‑CoV‑2 using reverse transcription loop‑mediated isothermal amplification (RT‑LAMP) Lena Mautner, Christin‑Kirsty Baillie, Heike Marie Herold, Wolfram Volkwein, Patrick Guertler, Ute Eberle, Nikolaus Ackermann, Andreas Sing, Melanie Pavlovic, Ottmar Goerlich, Ulrich Busch, Lars Wassill, Ingrid Huber and Armin Baiker*
Abstract Background: Fast, reliable and easy to handle methods are required to facilitate urgently needed point-of-care test‑ ing (POCT) in the current coronavirus pandemic. Life-threatening severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has rapidly spread all over the world, infecting more than 33,500,000 people and killing over 1 million of them as of October 2020. Infected individuals without any symptoms might still transfer the virus to others under‑ lining the extraordinary transmissibility of this new coronavirus. In order to identify early infections effectively, treat patients on time and control disease spreading, rapid, accurate and onsite testing methods are urgently required. Results: Here we report the development of a loop-mediated isothermal amplification (LAMP) based method to detect SARS-CoV-2 genes ORF8 and N directly from pharyngeal swab samples. The established reverse transcription LAMP (RT-LAMP) assay detects SARS-CoV-2 directly from pharyngeal swab samples without previous time-consuming and laborious RNA extraction. The assay is sensitive and highly specific for SARS-CoV-2 detection, showing no cross reactivity when tested on 20 other respiratory pathogens. The assay is 12 times faster and 10 times cheaper than rou‑ tine reverse transcription real-time polymerase chain reaction, depending on the assay used. Conclusion: The fast and easy to handle RT-LAMP assay amplifying specifically the genomic regions ORF8 and N of SARS-CoV-2 is ideally suited for POCT at e.g. railway stations, airports or hospitals. Given the current pandemic situa‑ tion, rapid, cost efficient and onsite methods like the here presented RT-LAMP assay are urgently needed to contain the viral spread. Keywords: RT-LAMP, Point-of-care testing, SARS-CoV-2, COVID-19, Rapid testing, No RNA extraction, ORF8, Gene N Introduction The emergence of the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has rapidly become a global public health emergency since its outbreak in Wuhan, China in December 2019 [1]. SARS-CoV-2 is an enveloped, non-segmented, single-stranded, positive-sense
*Correspondence: [email protected] Bavarian Health and Food Safety Authority, Veterinaerstrasse 2, 85764 Oberschleißheim, Germany
RNA virus similar to severe acute respiratory syndrome coronavirus (SARS-CoV) and Middle East respiratory syndrome coronavirus (MERS-CoV) [1]. Epidemical data show that the virus has strong human-to-human transmission ability, and is spread mainly by droplets produced by coughing and sneezing. Even talking or simple breathing can be enough for transmission [2]. The lifethreatening respiratory infections caused by
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