tetano
Editor, Senior Moderator
Biosens Bioelectron
. 2022 Mar 31;208:114221.
doi: 10.1016/j.bios.2022.114221. Online ahead of print.
Split T7 promoter-based isothermal transcription amplification for one-step fluorescence detection of SARS-CoV-2 and emerging variants
Taehwi Yoon[SUP] 1 [/SUP], Jiye Shin[SUP] 1 [/SUP], Hyun-Jung Choi[SUP] 2 [/SUP], Ki Soo Park[SUP] 3 [/SUP]
Affiliations
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.
. 2022 Mar 31;208:114221.
doi: 10.1016/j.bios.2022.114221. Online ahead of print.
Split T7 promoter-based isothermal transcription amplification for one-step fluorescence detection of SARS-CoV-2 and emerging variants
Taehwi Yoon[SUP] 1 [/SUP], Jiye Shin[SUP] 1 [/SUP], Hyun-Jung Choi[SUP] 2 [/SUP], Ki Soo Park[SUP] 3 [/SUP]
Affiliations
- PMID: 35421842
- DOI: 10.1016/j.bios.2022.114221
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.