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, Yi Sun

*Corresponding author for this work

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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 (<30 min), affordable, highly robust at room temperature (>1 month), isothermal (42 °C) and user-friendly, making it an excellent assay for broad-based testing.

Original languageEnglish
Article number5089
JournalNature Communications
Volume12
Issue number1
Number of pages12
ISSN2041-1723
DOIs
Publication statusPublished - 2021

Bibliographical note

Funding Information:
This work is financially supported by European Institute of Innovation & Technology (EIT) Health, Project no. 20876.

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