Abstract
BackgroundFast, reliable and easy to handle methods are required to facilitate urgently needed point-of-care testing (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 underlining 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.ResultsHere 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 routine reverse transcription real-time polymerase chain reaction, depending on the assay used.ConclusionThe 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 situation, rapid, cost efficient and onsite methods like the here presented RT-LAMP assay are urgently needed to contain the viral spread.
Highlights
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]
We report a novel reverse transcription loop-mediated isothermal amplification (RT-LAMP) detecting SARS-CoV-2 reliably in clinical swab samples without a laborious RNA extraction step
Optimising the Reverse transcription (RT)‐LAMP assay To screen for the optimum temperature of the designated reverse transcription LAMP (RT-LAMP), the reaction was incubated at three different temperatures (63 °C, 65 °C, 67 °C) for 35 min
Summary
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]. The lifethreatening respiratory infections caused by this virus designated as corona virus disease 2019 (COVID-19) are spreading worldwide [3]. Typical clinical symptoms of COVID-19 patients are fever, dry cough, breathing. 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
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