A non-enzymatic, isothermal strand displacement and amplification assay for rapid detection of SARS-CoV-2 RNA

Mohsen Mohammadniaei (), Ming Zhang, Jon Ashley, Ulf Bech Christensen, Lennart Jan Friis-Hansen, Rasmus Gregersen, Jan Gorm Lisby, Thomas Lars Benfield, Finn Erland Nielsen, Jens Henning Rasmussen, Ellen Bøtker Pedersen, Anne Christine Rye Olinger, Lærke Tørring Kolding, Maryam Naseri, Tao Zheng, Wentao Wang, Jan Gorodkin and Yi Sun ()
Additional contact information
Mohsen Mohammadniaei: Technical University of Denmark
Ming Zhang: Technical University of Denmark
Jon Ashley: Technical University of Denmark
Ulf Bech Christensen: PentaBase A/S
Lennart Jan Friis-Hansen: Copenhagen University Hospital - Bispebjerg and Frederiksberg
Rasmus Gregersen: Copenhagen University Hospital - Bispebjerg and Frederiksberg
Jan Gorm Lisby: Copenhagen University Hospital - Amager and Hvidovre
Thomas Lars Benfield: Copenhagen University Hospital - Amager and Hvidovre
Finn Erland Nielsen: Copenhagen University Hospital - Bispebjerg and Frederiksberg
Jens Henning Rasmussen: Copenhagen University Hospital - Bispebjerg and Frederiksberg
Ellen Bøtker Pedersen: Copenhagen University Hospital - Bispebjerg and Frederiksberg
Anne Christine Rye Olinger: Copenhagen University Hospital - Bispebjerg and Frederiksberg
Lærke Tørring Kolding: Copenhagen University Hospital - Bispebjerg and Frederiksberg
Maryam Naseri: Technical University of Denmark
Tao Zheng: Technical University of Denmark
Wentao Wang: Technical University of Denmark
Jan Gorodkin: University of Copenhagen
Yi Sun: Technical University of Denmark

Nature Communications, 2021, vol. 12, issue 1, 1-12

Abstract: Abstract The current nucleic acid signal amplification methods for SARS-CoV-2 RNA detection heavily rely on the functions of biological enzymes which imposes stringent transportation and storage conditions, high cost and global supply shortages. Here, a non-enzymatic whole genome detection method based on a simple isothermal signal amplification approach is developed for rapid detection of SARS-CoV-2 RNA and potentially any types of nucleic acids regardless of their size. The assay, termed non-enzymatic isothermal strand displacement and amplification (NISDA), is able to quantify 10 RNA copies.µL−1. In 164 clinical oropharyngeal RNA samples, NISDA assay is 100 % specific, and it is 96.77% and 100% sensitive when setting up in the laboratory and hospital, respectively. The NISDA assay does not require RNA reverse-transcription step and is fast ( 1 month), isothermal (42 °C) and user-friendly, making it an excellent assay for broad-based testing.

Date: 2021
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DOI: 10.1038/s41467-021-25387-9

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