Portable, single nucleotide polymorphism-specific duplex assay for virus surveillance in wastewater

Portable, single nucleotide polymorphism-specific duplex assay for virus surveillance in wastewater

The Argonaute protein from the archaeon Pyrococcus furiosus (PfAgo) is a DNA-guided nuclease that targets DNA with any sequence. We designed a virus detection assay in which the PfAgo enzyme cleaves the reporter probe, thus generating fluorescent signals when amplicons from a reverse transcriptase l...

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Veröffentlicht in:The Science of the total environment 2024-02, Vol.912, p.168701-168701, Article 168701
Hauptverfasser: Oh, Chamteut, Xun, Guanhua, Lane, Stephan Thomas, Petrov, Vassily Andrew, Zhao, Huimin, Nguyen, Thanh H.
Format: Artikel
Sprache:eng
Schlagworte:
confidence interval
detection limit
Diagnostic assay
DNA
environment
enzymes
fluorescence
monitoring
Pepper mild mottle virus
PfAgo
Portable
Pyrococcus furiosus
reverse transcription loop-mediated isothermal amplification
Severe acute respiratory syndrome coronavirus 2
single nucleotide polymorphism
Single nucleotide polymorphism detection
Virus
viruses
Wastewater
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Beschreibung
Zusammenfassung:The Argonaute protein from the archaeon Pyrococcus furiosus (PfAgo) is a DNA-guided nuclease that targets DNA with any sequence. We designed a virus detection assay in which the PfAgo enzyme cleaves the reporter probe, thus generating fluorescent signals when amplicons from a reverse transcriptase loop-mediated isothermal amplification (RT-LAMP) assay contain target sequences. We confirmed that the RT-LAMP-PfAgo assay for the SARS-CoV-2 Delta variant produced significantly higher fluorescent signals (p 
ISSN:0048-9697
1879-1026
DOI:10.1016/j.scitotenv.2023.168701