Rapid point-of-care detection of SARS-CoV-2 using reverse transcription loop-mediated isothermal amplification (RT-LAMP).
Virol J. 2020 10 21; 17(1):160.VJ

Abstract

BACKGROUND

Fast, 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.

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 routine 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 situation, rapid, cost efficient and onsite methods like the here presented RT-LAMP assay are urgently needed to contain the viral spread.

Links

Publisher Full Text
ncbi.nlm.nih.gov
virologyj.biomedcentral.com
PMC Free PDF

Authors+Show Affiliations

Mautner L
Bavarian Health and Food Safety Authority, Veterinaerstrasse 2, 85764, Oberschleiβheim, Germany.
Baillie CK
Bavarian Health and Food Safety Authority, Veterinaerstrasse 2, 85764, Oberschleiβheim, Germany.
Herold HM
Bavarian Health and Food Safety Authority, Veterinaerstrasse 2, 85764, Oberschleiβheim, Germany.
Volkwein W
Bavarian Health and Food Safety Authority, Veterinaerstrasse 2, 85764, Oberschleiβheim, Germany.
Guertler P
Bavarian Health and Food Safety Authority, Veterinaerstrasse 2, 85764, Oberschleiβheim, Germany.
Eberle U
Bavarian Health and Food Safety Authority, Veterinaerstrasse 2, 85764, Oberschleiβheim, Germany.
Ackermann N
Bavarian Health and Food Safety Authority, Veterinaerstrasse 2, 85764, Oberschleiβheim, Germany.
Sing A
Bavarian Health and Food Safety Authority, Veterinaerstrasse 2, 85764, Oberschleiβheim, Germany.
Pavlovic M
Bavarian Health and Food Safety Authority, Veterinaerstrasse 2, 85764, Oberschleiβheim, Germany.
Goerlich O
Bavarian Health and Food Safety Authority, Veterinaerstrasse 2, 85764, Oberschleiβheim, Germany.
Busch U
Bavarian Health and Food Safety Authority, Veterinaerstrasse 2, 85764, Oberschleiβheim, Germany.
Wassill L
Bavarian Health and Food Safety Authority, Veterinaerstrasse 2, 85764, Oberschleiβheim, Germany.
Huber I
Bavarian Health and Food Safety Authority, Veterinaerstrasse 2, 85764, Oberschleiβheim, Germany.
Baiker A
Bavarian Health and Food Safety Authority, Veterinaerstrasse 2, 85764, Oberschleiβheim, Germany. armin.baiker@lgl.bayern.de.

MeSH

AnimalsBetacoronavirusCOVID-19COVID-19 TestingChlorocebus aethiopsClinical Laboratory TechniquesCoronavirus InfectionsGenes, ViralHumansMolecular Diagnostic TechniquesNucleic Acid Amplification TechniquesPandemicsPneumonia, ViralPoint-of-Care SystemsRNA, ViralReal-Time Polymerase Chain ReactionReverse TranscriptionSARS-CoV-2Vero Cells

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

33087160