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Dataset for: Ultrarapid detection of SARS-CoV-2 RNA using a reverse transcription-free exponential amplification reaction, RTF-EXPAR

Citation

Carter, Jake G. et al. (2022), Dataset for: Ultrarapid detection of SARS-CoV-2 RNA using a reverse transcription-free exponential amplification reaction, RTF-EXPAR, Dryad, Dataset, https://doi.org/10.5061/dryad.k0p2ngf8s

Abstract

A dataset is reported for a rapid isothermal method for detecting severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), the virus responsible for COVID-19. The procedure uses an unprecedented reverse transcription–free (RTF) approach for converting genomic RNA into DNA. This involves the formation of an RNA/DNA heteroduplex whose selective cleavage generates a short DNA trigger strand, which is then rapidly amplified using the exponential amplification reaction (EXPAR). Deploying the RNA-to-DNA conversion and amplification stages of the RTF-EXPAR assay in a single step results in the detection, via a fluorescence read-out, of single figure copy numbers per microliter of SARS-CoV-2 RNA in under 10 min. In direct three-way comparison studies the assay has been found to be faster than both PCR and loop-mediated isothermal amplification (LAMP), while being just as sensitive. The assay protocol involves the use of standard laboratory equipment and is readily adaptable for the detection of other RNA-based agents.

Methods

Fluorescence data was collected and exported as a plain text file for data processing (primary data files). Normalised data was determined using normalise function in GraphPad Prism 9 (normalised data files). To analyze the data, a program in C# was developed (cs format files). Each run time was calculated to be the point at which the fluorescence signal was greater than 10 SDs from the baseline signal (10-sigma time).

Usage Notes

No additional information is required but corresponding authors (Dafforn and Tucker) can be contacted if neccesary.

Funding

Biotechnology and Biological Sciences Research Council, Award: BB/R506175/1