Abstract
The negative global impact of the coronavirus disease pandemic has highlighted the crucial need for a rapid and convenient method of viral RNA detection. In this study, we report a novel method, termed as the split T7 promoter-based isothermal transcription amplification with light-up RNA aptamer (STAR), for one-pot detection of viral RNA. STAR uses a split T7 promoter that is applied to a three-way junction to mediate the selective transcription by the T7 RNA polymerase in the presence of target RNA. In addition, a light-up RNA aptamer is used for signal amplification. STAR can detect viral RNA in less than 30 min with high specificity and sensitivity. By testing of 60 nasopharyngeal SARS-CoV-2 samples, the STAR assay demonstrates an excellent sensitivity and specificity of 96.7% and 100%, respectively. Moreover, we provide experimental evidence of the broad applicability of this assay through the multiplex detection of SARS-CoV-2 variants (D614G mutation) and direct detection of bacterial 16S rRNA.
Keywords: Isothermal amplification; Light-up RNA aptamer; SARS-CoV-2; Split T7 promoter; Three-way junction; Transcription.
【저자키워드】 SARS-CoV-2, isothermal amplification, Light-up RNA aptamer, Split T7 promoter, Three-way junction, Transcription., 【초록키워드】 coronavirus disease, pandemic, Transcription, SARS-CoV-2 variant, RNA, 16S rRNA, amplification, sensitivity, specificity, D614G mutation, Sensitivity and specificity, SARS-CoV-2 variants, nasopharyngeal, isothermal amplification, Viral RNA, multiplex, rRNA, Bacterial, isothermal, evidence of, STAR, selective, applicability, split, promoter, viral RNA detection, experimental evidence, T7 RNA polymerase, detect, addition, applied, less, 【제목키워드】 variant, amplification, fluorescence detection, isothermal, split,