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

2021; National Academy of Sciences; Volume: 118; Issue: 35 Linguagem: Inglês

10.1073/pnas.2100347118

1091-6490

Jake G. Carter, Lorea Orueta Iturbe, Jean‐Louis H. A. Duprey, Ian R. Carter, Craig D. Southern, Marium Rana, Celina Whalley, Andrew Bosworth, Andrew D. Beggs, Matthew R. Hicks, James H. R. Tucker, Timothy R. Dafforn,

Advanced biosensing and bioanalysis techniques

Significance We report a rapid COVID-19 assay that gives a sample-to-signal time of under 10 min. The current gold-standard COVID-19 assay uses PCR, where strands of DNA are copied (amplified) many times to generate a read-out signal. However, as the virus genome is RNA, first conversion into DNA is required using reverse transcription (RT) before amplification. While just as sensitive, our assay is faster because 1) we have designed a method for generating DNA (the trigger strand) from RNA, bypassing the lengthy RT step, and 2) a quicker amplification process than PCR, called exponential amplification reaction (EXPAR), is used to amplify the trigger. This methodology could ultimately be applied to any RNA-based assay, including the detection of other infectious agents.