Elsevier

Talanta

Volume 252, 15 January 2023, 123835
Talanta

Multiple ligation–Assisted recombinase polymerase amplification for highly sensitive and selective colorimetric detection of SARS-CoV-2

https://doi.org/10.1016/j.talanta.2022.123835Get rights and content

Abstract

In this paper we present a new method for the detection of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), targeting a specific region “N gene.” Under isothermal reaction conditions, we integrated ligation (Lig; high selectivity) and recombinase polymerase amplification (RPA; high sensitivity) processes, obtaining a robust method of detection. For point-of-care testing, we incorporated our laboratory-produced pyrophosphate ion (PPi)–sensing probe (PK-probe) for colorimetric analysis of the reaction. The total detection system was efficient and effective at diagnosing this RNA virus–mediated disease rapidly (30 min). In a full-genome SARS-CoV-2 study, our PK-probe/Lig-RPA system functioned with a limit of detection of 1160 copies/ml, with a single-mismatch level of selectively, and it was highly selective even in the presence of bacterial genomes commonly found in the human mouth and nose. This robust, straightforward, selective, efficient, and ultrasensitive colorimetric detection method, with potential for point-of-care analysis, should also be effective in detecting a diverse range of other RNA-based diseases.

Graphical abstract

We developed a new method named as PK-probe/Lig-RPA assay for the detection of SARS-CoV-2 targeting a specific “N gene” region.

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Keywords

Pyrophosphate
Colorimetric sensing
PK-probe
Isothermal amplification
Recombinase polymerase amplification (RPA)
SARS-CoV-2

Data availability

The data that has been used is confidential.

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