Split T7 promoter-based isothermal transcription amplification for one-step fluorescence detection of SARS-CoV-2 and emerging variants

• Main Authors: Choi, H.-J (Author), Park, K.S (Author), Shin, J. (Author), Yoon, T. (Author)
• Format: Article
• Language: English
• Published: Elsevier Ltd 2022
• Subjects: Amplification; Coronaviruses; Diseases; Fluorescence detection; Global impacts; Isothermal amplification; Isothermal amplifications; Isotherms; Light-up RNA aptamer; RNA; RNA aptamer; SARS; SARS-CoV-2; Split t7 promoter; Split T7 promoter; Stars; Three-way junction; Transcription; Viral RNA
• Online Access: View Fulltext in Publisher

 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. © 2022 Elsevier B.V.