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

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...

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Detalles Bibliográficos
Autores principales: Mohammadniaei, Mohsen, Zhang, Ming, Ashley, Jon, Christensen, Ulf Bech, Friis-Hansen, Lennart Jan, Gregersen, Rasmus, Lisby, Jan Gorm, Benfield, Thomas Lars, Nielsen, Finn Erland, Henning Rasmussen, Jens, Pedersen, Ellen Bøtker, Olinger, Anne Christine Rye, Kolding, Lærke Tørring, Naseri, Maryam, Zheng, Tao, Wang, Wentao, Gorodkin, Jan, Sun, Yi
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Nature Publishing Group UK 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8385016/
https://www.ncbi.nlm.nih.gov/pubmed/34429424
http://dx.doi.org/10.1038/s41467-021-25387-9
Descripción
Sumario: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.