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

被引：39

作者：

Yoon, Taehwi ^{[1
]}

Shin, Jiye ^{[1
]}

Choi, Hyun-Jung ^{[2
,3
]}

Park, Ki Soo ^{[1
]}

机构：

[1] Konkuk Univ, Coll Engn, Dept Biol Engn, Seoul, South Korea

[2] Chonnam Natl Univ, Dept Lab Med, Med Sch, Gwangju, South Korea

[3] Chonnam Natl Univ Hosp, Gwangju, South Korea

来源：

BIOSENSORS & BIOELECTRONICS | 2022年 / 208卷

基金：

新加坡国家研究基金会;

关键词：

SARS-CoV-2; Isothermal amplification; Split T7 promoter; Three-way junction; Transcription; Light-up RNA aptamer; DNA;

D O I：

10.1016/j.bios.2022.114221

中图分类号：

Q6 [生物物理学];

学科分类号：

071011 ;

摘要：

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.

引用

页数：8

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