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Microfluidic toolbox using padlock probes and rolling circle amplification for direct detection and genotyping of viral RNA
KTH, School of Engineering Sciences in Chemistry, Biotechnology and Health (CBH), Protein Science, Nano Biotechnology. KTH, Centres, Science for Life Laboratory, SciLifeLab. AIMES (Center for the Advancement of Integrated Medical and Engineering Sciences), Karolinska Institutet, Stockholm, Sweden.
ICBAS-School of Medicine and Biomedical Sciences, University of Porto, Porto, Portugal; Epidemiology Research Unit (EPIunit), Institute of Public Health, University of Porto, Porto, Portugal; Laboratório para a Investigação Integrativa e Translacional em Saúde Populacional (ITR), Porto, Portugal.ORCID iD: 0000-0002-8384-0640
Department of Biochemistry and Biophysics, Science for Life Laboratory, Stockholm University, Solna, Sweden.
ICBAS-School of Medicine and Biomedical Sciences, University of Porto, Porto, Portugal; Epidemiology Research Unit (EPIunit), Institute of Public Health, University of Porto, Porto, Portugal; Laboratório para a Investigação Integrativa e Translacional em Saúde Populacional (ITR), Porto, Portugal.
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2026 (English)In: RSC Advances, E-ISSN 2046-2069, Vol. 16, no 21, p. 19119-19127Article in journal (Refereed) Published
Abstract [en]

Rolling circle amplification (RCA) combined with padlock probes presents a promising tool for direct detection and genotyping of viral RNA, offering advantages over conventional methods like RT-PCR. This isothermal process enables highly sensitive and specific amplification of nucleic acids without the need for thermal cycling, making it suitable for point-of-care applications. In this study, we demonstrate a microfluidic and RCA-based method for the direct detection of SARS-CoV-2 RNA and variant profiling, bypassing the reverse transcription step. Our approach allows for the identification of single nucleotide polymorphisms (SNPs) specific to viral variants, enhancing the detection sensitivity through the circle-to-circle amplification (C2CA) technique. This methodology shows potential as a robust, cost-effective platform for viral diagnostics, capable of being fully automated and integrated with miniaturized detection systems for efficient use in both resource-rich and resource-limited settings.

Place, publisher, year, edition, pages
Royal Society of Chemistry (RSC) , 2026. Vol. 16, no 21, p. 19119-19127
National Category
Molecular Biology Medical Genetics and Genomics Infectious Medicine
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URN: urn:nbn:se:kth:diva-381094DOI: 10.1039/d6ra00912cISI: 001736682700001PubMedID: 41969386Scopus ID: 2-s2.0-105035880604OAI: oai:DiVA.org:kth-381094DiVA, id: diva2:2059095
Note

QC 20260511

Available from: 2026-05-11 Created: 2026-05-11 Last updated: 2026-05-11Bibliographically approved

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Varela, João C.Russom, Aman

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Varela, João C.Gomes da Silva, PriscillaRussom, AmanSoares, Ruben R.G.
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