Ultra-fast detection and quantification of nucleic acids by amplification-free fluorescence assay

Publikation: Bidrag til tidsskrift/Konferencebidrag i tidsskrift /Bidrag til avisTidsskriftartikelForskningpeer review

DOI

  • Jesper Uhd, Danmarks Tekniske Universitet
  • ,
  • Laura Miotke, Stanford University
  • ,
  • Hanlee P. Ji, Stanford University
  • ,
  • Marina Dunaeva, Radboud University Nijmegen
  • ,
  • Ger J.M. Pruijn, Radboud University Nijmegen
  • ,
  • Christian Damsgaard Jørgensen, Department ofMathematics and Computer Science, Syddansk Universitet
  • ,
  • Emil Laust Kristoffersen
  • Victoria Birkedal
  • Christina Westmose Yde, Københavns Universitet
  • ,
  • Finn Cilius Nielsen
  • Jonas Hansen, Institute of Molecular Medicine, Sechenov First Moscow State Medical University
  • ,
  • Kira Astakhova, Danmarks Tekniske Universitet

Two types of clinically important nucleic acid biomarkers, microRNA (miRNA) and circulating tumor DNA (ctDNA) were detected and quantified from human serum using an amplification-free fluorescence hybridization assay. Specifically, miRNAs hsa-miR-223-3p and hsa-miR-486-5p with relevance for rheumatoid arthritis and cancer related mutations BRAF and KRAS of ctDNA were directly measured. The required oligonucleotide probes for the assay were rationally designed and synthesized through a novel "clickable"approach which is time and cost-effective. With no need for isolating nucleic acid components from serum, the fluoresence-based assay took only 1 hour. Detection and absolute quantification of targets was successfully achieved despite their notoriously low abundance, with a precision down to individual nucleotides. Obtained miRNA and ctDNA amounts showed overall a good correlation with current techniques. With appropriate probes, our novel assay and signal boosting approach could become a useful tool for point-of-care measuring other low abundance nucleic acid biomarkers.

OriginalsprogEngelsk
TidsskriftAnalyst
Vol/bind145
Nummer17
Sider (fra-til)5836-5844
ISSN0003-2654
DOI
StatusUdgivet - sep. 2020

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