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

Jesper Uhd, Laura Miotke, Hanlee P Ji, Marina Dunaeva, Ger J M Pruijn, Christian Damsgaard Jørgensen, Emil Laust Kristoffersen, Victoria Birkedal, Christina Westmose Yde, Finn Cilius Nielsen, Jonas Hansen, Kira Astakhova*

*Corresponding author for this work

Research output: Contribution to journalJournal articleResearchpeer-review


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.

Original languageEnglish
JournalThe Analyst
Pages (from-to)5836-5844
Publication statusPublished - 2020


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