Universität zu Köln

Schermer, Bernhard, Fabretti, Francesca, Damagnez, Maximilian ORCID: 0000-0002-2745-7719, Di Cristanziano, Veronica, Heger, Eva, Arjune, Sita ORCID: 0000-0002-6121-4614, Tanner, Nathan A., Imhof, Thomas, Koch, Manuel, Ladha, Alim, Joung, Julia, Gootenberg, Jonathan S., Abudayyeh, Omar O., Burst, Volker, Zhang, Feng, Klein, Florian, Benzing, Thomas and Mueller, Roman-Ulrich (2020). Rapid SARS-CoV-2 testing in primary material based on a novel multiplex RT-LAMP assay. PLoS One, 15 (11). SAN FRANCISCO: PUBLIC LIBRARY SCIENCE. ISSN 1932-6203

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Abstract

Background Rapid and extensive testing of large parts of the population and specific subgroups is crucial for proper management of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infections and decision-making in times of a pandemic outbreak. However, point-of-care (POC) testing in places such as emergency units, outpatient clinics, airport security points or the entrance of any public building is a major challenge. The need for thermal cycling and nucleic acid isolation hampers the use of standard PCR-based methods for this purpose. Methods To avoid these obstacles, we tested PCR-independent methods for the detection of SARS-CoV-2 RNA from primary material (nasopharyngeal swabs) including reverse transcription loop-mediated isothermal amplification (RT-LAMP) and specific high-sensitivity enzymatic reporter unlocking (SHERLOCK). Results Whilst specificity of standard RT-LAMP assays appears to be satisfactory, sensitivity does not reach the current gold-standard quantitative real-time polymerase chain reaction (qPCR) assays yet. We describe a novel multiplexed RT-LAMP approach and validate its sensitivity on primary samples. This approach allows for fast and reliable identification of infected individuals. Primer optimization and multiplexing helps to increase sensitivity significantly. In addition, we directly compare and combine our novel RT-LAMP assays with SHERLOCK. Conclusion In summary, this approach reveals one-step multiplexed RT-LAMP assays as a prime-option for the development of easy and cheap POC test kits.

Item Type:	Journal Article
Creators:	Creators Email ORCID ORCID Put Code Schermer, Bernhard UNSPECIFIED UNSPECIFIED UNSPECIFIED Fabretti, Francesca UNSPECIFIED UNSPECIFIED UNSPECIFIED Damagnez, Maximilian UNSPECIFIED orcid.org/0000-0002-2745-7719 UNSPECIFIED Di Cristanziano, Veronica UNSPECIFIED UNSPECIFIED UNSPECIFIED Heger, Eva UNSPECIFIED UNSPECIFIED UNSPECIFIED Arjune, Sita UNSPECIFIED orcid.org/0000-0002-6121-4614 UNSPECIFIED Tanner, Nathan A. UNSPECIFIED UNSPECIFIED UNSPECIFIED Imhof, Thomas UNSPECIFIED UNSPECIFIED UNSPECIFIED Koch, Manuel UNSPECIFIED UNSPECIFIED UNSPECIFIED Ladha, Alim UNSPECIFIED UNSPECIFIED UNSPECIFIED Joung, Julia UNSPECIFIED UNSPECIFIED UNSPECIFIED Gootenberg, Jonathan S. UNSPECIFIED UNSPECIFIED UNSPECIFIED Abudayyeh, Omar O. UNSPECIFIED UNSPECIFIED UNSPECIFIED Burst, Volker UNSPECIFIED UNSPECIFIED UNSPECIFIED Zhang, Feng UNSPECIFIED UNSPECIFIED UNSPECIFIED Klein, Florian UNSPECIFIED UNSPECIFIED UNSPECIFIED Benzing, Thomas UNSPECIFIED UNSPECIFIED UNSPECIFIED Mueller, Roman-Ulrich UNSPECIFIED UNSPECIFIED UNSPECIFIED
URN:	urn:nbn:de:hbz:38-312361
DOI:	10.1371/journal.pone.0238612
Journal or Publication Title:	PLoS One
Volume:	15
Number:	11
Date:	2020
Publisher:	PUBLIC LIBRARY SCIENCE
Place of Publication:	SAN FRANCISCO
ISSN:	1932-6203
Language:	English
Faculty:	Unspecified
Divisions:	Unspecified
Subjects:	no entry
Uncontrolled Keywords:	Keywords Language MEDIATED ISOTHERMAL AMPLIFICATION Multiple languages Multidisciplinary Sciences Multiple languages
URI:	http://kups.ub.uni-koeln.de/id/eprint/31236