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An Immiscible-Phase Filtration Device for Isolation, Amplification, and Detection of Nucleic Acids for Clinical Diagnostics
Stockholm University, Faculty of Science, Department of Materials and Environmental Chemistry (MMK).ORCID iD: 0000-0002-7102-5024
Stockholm University, Faculty of Science, Department of Materials and Environmental Chemistry (MMK).ORCID iD: 0000-0002-9391-9387
Number of Authors: 32024 (English)In: Microfluidics Diagnostics: Methods and Protocols / [ed] Valérie Taly, Stéphanie Descroix, Karla Perez-Toralla, Humana Press, 2024, p. 195-206Chapter in book (Refereed)
Abstract [en]

Clinical diagnostics of infectious diseases via nucleic acid amplification tests (NAATs) depend on a separate step of isolation of nucleic acids from cells/viruses embedded in complex biological matrices. The most recent example has been reverse transcription polymerase chain reaction (RT-PCR) for amplification and detection of SARS-CoV-2 RNA for COVID-19 diagnostics. Kits for RNA extraction and purification are commercially available; however, their integration with amplification systems is generally lacking, resulting in two separate steps, i.e., sample preparation and amplification. This makes NAATs more time-consuming, requiring skilled personnel, and can increase the likelihood of contamination. Here, we describe a setup and methodology to perform the quick extraction and detection of nucleic acids in an integrated manner. In particular, we focus on the use of an immiscible filtration device for capture, isolation, concentration, amplification, and colorimetric detection of SARS-CoV-2 RNA.

Place, publisher, year, edition, pages
Humana Press, 2024. p. 195-206
Series
Methods in Molecular Biology, ISSN 1064-3745, E-ISSN 1940-6029 ; 2804
Keywords [en]
COVID-19, Diagnostics, Extraction, IFAST, Immiscible filtration, Isothermal amplification
National Category
Biochemistry Molecular Biology
Identifiers
URN: urn:nbn:se:su:diva-236113DOI: 10.1007/978-1-0716-3850-7_12PubMedID: 38753149Scopus ID: 2-s2.0-85193384705ISBN: 9781071638491 (print)OAI: oai:DiVA.org:su-236113DiVA, id: diva2:1917401
Available from: 2024-12-02 Created: 2024-12-02 Last updated: 2025-02-20Bibliographically approved

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Rodriguez Mateos, PabloPamme, Nicole

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