逆转录酶
核糖核酸
异源双工
逆转录环介导等温扩增
环介导等温扩增
分子生物学
重组酶聚合酶扩增
DNA
多重位移放大
生物
抄写(语言学)
化学
病毒学
聚合酶链反应
基因
遗传学
DNA提取
哲学
语言学
作者
Jake G. Carter,Lorea Orueta Iturbe,Jean‐Louis H. A. Duprey,Ian R. Carter,Craig D. Southern,Marium Rana,Celina M. Whalley,Andrew Bosworth,Andrew D Beggs,Matthew R. Hicks,James H. R. Tucker,TR Dafforn
标识
DOI:10.1073/pnas.2100347118
摘要
A rapid isothermal method for detecting severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), the virus responsible for COVID-19, is reported. 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 RT-qPCR and reverse transcription loop-mediated isothermal amplification (RT-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 pathogens.
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