tetano
Editor, Senior Moderator
Biopolymers
. 2022 Jan 13;e23485.
doi: 10.1002/bip.23485. Online ahead of print.
Combining recombinase polymerase amplification and DNA-templated reaction for SARS-CoV-2 sensing with dual fluorescence and lateral flow assay output
Lluc Farrera-Soler[SUP] 1 [/SUP], Arthur Gonse[SUP] 1 [/SUP], Ki Tae Kim[SUP] 1 [/SUP], Sofia Barluenga[SUP] 1 [/SUP], Nicolas Winssinger[SUP] 1 [/SUP]
Affiliations
Abstract
The early phase of the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) pandemic was exacerbated by a diagnostic challenge of unprecedented magnitude. In the absence of effective therapeutics or vaccines, breaking the chain of transmission through early disease detection and patient isolation was the only means to control the growing pandemic. While polymerase chain reaction (PCR)-based methods and rapid-antigen tests rose to the occasion, the analytical challenge of rapid and sequence-specific nucleic acid-sensing at a point-of-care or home setting stimulated intense developments. Herein we report a method that combines recombinase polymerase amplification and a DNA-templated reaction to achieve a dual readout with either fluorescence (microtiter plate) or naked eye (lateral flow assay: LFA) detection. The nucleic acid templated reaction is based on an S[SUB]N[/SUB] Ar that simultaneously transfers biotin from one Peptide Nucleic Acid (PNA) strand to another PNA strand, enabling LFA detection while uncaging a coumarin for fluorescence readout. This methodology has been applied to the detection of a DNA or RNA sequence uniquely attributed to the SARS-CoV-2.
Keywords: DNA templated reaction; PNA; RPA; lateral flow assay; nucleic acid sensing.
. 2022 Jan 13;e23485.
doi: 10.1002/bip.23485. Online ahead of print.
Combining recombinase polymerase amplification and DNA-templated reaction for SARS-CoV-2 sensing with dual fluorescence and lateral flow assay output
Lluc Farrera-Soler[SUP] 1 [/SUP], Arthur Gonse[SUP] 1 [/SUP], Ki Tae Kim[SUP] 1 [/SUP], Sofia Barluenga[SUP] 1 [/SUP], Nicolas Winssinger[SUP] 1 [/SUP]
Affiliations
- PMID: 35023571
- DOI: 10.1002/bip.23485
Abstract
The early phase of the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) pandemic was exacerbated by a diagnostic challenge of unprecedented magnitude. In the absence of effective therapeutics or vaccines, breaking the chain of transmission through early disease detection and patient isolation was the only means to control the growing pandemic. While polymerase chain reaction (PCR)-based methods and rapid-antigen tests rose to the occasion, the analytical challenge of rapid and sequence-specific nucleic acid-sensing at a point-of-care or home setting stimulated intense developments. Herein we report a method that combines recombinase polymerase amplification and a DNA-templated reaction to achieve a dual readout with either fluorescence (microtiter plate) or naked eye (lateral flow assay: LFA) detection. The nucleic acid templated reaction is based on an S[SUB]N[/SUB] Ar that simultaneously transfers biotin from one Peptide Nucleic Acid (PNA) strand to another PNA strand, enabling LFA detection while uncaging a coumarin for fluorescence readout. This methodology has been applied to the detection of a DNA or RNA sequence uniquely attributed to the SARS-CoV-2.
Keywords: DNA templated reaction; PNA; RPA; lateral flow assay; nucleic acid sensing.