Abstract
Nucleic acid-based detection of RNA viruses requires an annealing procedure to obtain RNA/probe or RNA/primer complexes for unwinding stable structures of folded viral RNA. In this study, we designed a protein-enzyme-free nano-construction, named four-armed DNA machine (4DNM), that requires neither an amplification stage nor a high-temperature annealing step for SARS-CoV-2 detection. It uses a binary deoxyribozyme (BiDz) sensor incorporated in a DNA nanostructure equipped with a total of four RNA-binding arms. Additional arms were found to improve the limit of detection at least 10-fold. The sensor distinguished SARS-CoV-2 from other respiratory viruses and correctly identified five positive and six negative clinical samples verified by quantitative polymerase chain reaction (RT-qPCR). The strategy reported here can be used for the detection of long natural RNA and can become a basis for a point-of-care or home diagnostic test.
Keywords: 10-23 DNAzyme; DNA machine; binary hybridization probe; deoxyribozyme sensor; detection of folded RNA.
【저자키워드】 detection of folded RNA., deoxyribozyme sensor, binary hybridization probe, DNA machine, 10-23 DNAzyme, 【초록키워드】 Structure, SARS-CoV-2, diagnostic, RNA, point-of-care, amplification, DNA, SARS-CoV-2 detection, RT-qPCR, limit of detection, Viral RNA, Quantitative, ARMS, positive, Arm, FIVE, polymerase chain, annealing, IMPROVE, clinical sample, reported, can be used, RNA virus, complexes, other respiratory virus, 【제목키워드】 COVID-19, SARS-CoV-2, Test, detection, DNA, clinical, machine, home, Sample,