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

doi:10.1039/d0an00676a

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 af dette arbejde

Publikation: Bidrag til tidsskrift/Konferencebidrag i tidsskrift /Bidrag til avis › Tidsskriftartikel › Forskning › peer review

9 Citationer (Scopus)

Abstract

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.

Originalsprog	Engelsk
Tidsskrift	Analyst
Vol/bind	145
Nummer	17
Sider (fra-til)	5836-5844
ISSN	0003-2654
DOI	https://doi.org/10.1039/d0an00676a
Status	Udgivet - sep. 2020

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Citationsformater

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abstract = "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.",

author = "Jesper Uhd and Laura Miotke and Ji, \{Hanlee P.\} and Marina Dunaeva and Pruijn, \{Ger J.M.\} and J{\o}rgensen, \{Christian Damsgaard\} and Kristoffersen, \{Emil Laust\} and Victoria Birkedal and Yde, \{Christina Westmose\} and Nielsen, \{Finn Cilius\} and Jonas Hansen and Kira Astakhova",

year = "2020",

month = sep,

doi = "10.1039/d0an00676a",

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volume = "145",

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T1 - Ultra-fast detection and quantification of nucleic acids by amplification-free fluorescence assay

AU - Uhd, Jesper

AU - Miotke, Laura

AU - Ji, Hanlee P.

AU - Dunaeva, Marina

AU - Pruijn, Ger J.M.

AU - Jørgensen, Christian Damsgaard

AU - Kristoffersen, Emil Laust

AU - Birkedal, Victoria

AU - Yde, Christina Westmose

AU - Nielsen, Finn Cilius

AU - Hansen, Jonas

AU - Astakhova, Kira

PY - 2020/9

Y1 - 2020/9

N2 - 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.

AB - 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.

UR - http://www.scopus.com/inward/record.url?scp=85089815572&partnerID=8YFLogxK

U2 - 10.1039/d0an00676a

DO - 10.1039/d0an00676a

M3 - Journal article

C2 - 32648858

AN - SCOPUS:85089815572

SN - 0003-2654

VL - 145

SP - 5836

EP - 5844

JO - Analyst

JF - Analyst

IS - 17

ER -

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

Abstract

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