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. The reliance on enzymes in SARS-CoV-2 RNA detection imposes limits on transport and storage conditions. Here the authors use non-enzymatic isothermal amplification to detect RNA with no need for reverse transcription.
【저자키워드】 SARS-CoV-2, nucleic acids, Assay systems, 【초록키워드】 hospital, Laboratory, RNA, amplification, nucleic acid, nucleic acids, temperature, reverse transcription, isothermal amplification, SARS-CoV-2 RNA, Oropharyngeal, Enzymes, function, SARS-CoV-2 RNA detection, isothermal, room temperature, Transport, Supply shortages, enzyme, RNA samples, limits, functions, whole genome, storage conditions, approach, limit, robust, detect, 【제목키워드】 Amplification assay,