| Title: |
Amplification-free nucleic acid detection with a fluorescence-based waveguide biosensor |
| Authors: |
Philip A. Kocheril, Kiersten D. Lenz, Daniel E. Jacobsen, Jessica Z. Kubicek-Sutherland |
| Source: |
Frontiers in Sensors, Vol 3 (2022) |
| Publisher Information: |
Frontiers Media S.A., 2022. |
| Publication Year: |
2022 |
| Collection: |
LCC:Biotechnology |
| Subject Terms: |
waveguide, fluorescence spectroscopy, nucleic acid, amplification-free, molecular beacon, langmuir adsorption, Biotechnology, TP248.13-248.65 |
| More Details: |
Early detection of pathogens using nucleic acids in clinical samples often requires sensitivity at the single-copy level, which currently necessitates time-consuming and expensive nucleic acid amplification. Here, we describe 1) a redesigned flow cell in the shape of a trapezoid-subtracted geometric stadium, and 2) modified experimental procedures that allow for the measurement of sub-attomolar analytes in microliter quantities on a fluorescence-based waveguide biosensor. We verified our instrumental sensitivity with a 200-μL sample of a fluorescent streptavidin conjugate at 100 zM (100 zeptomolar, or 100·10−21 mol L−1) and theoretically explored the applicability of this modified sensing platform in a sandwich immunoassay format using a Langmuir adsorption model. We present assays that demonstrate specific detection of synthetic influenza A DNA (in buffer) and RNA (in saliva) oligonucleotides at the single-copy level (200 μL at 10 zM) using a fluorescent molecular beacon. Lastly, we demonstrate detection of isolated genomic influenza A RNA at a clinically relevant concentration. This work constitutes a sensitivity improvement of over twelve orders of magnitude compared to our previous nucleic acid detection work, illustrating the significant enhancements that can be gained with optimized experimental design. |
| Document Type: |
article |
| File Description: |
electronic resource |
| Language: |
English |
| ISSN: |
2673-5067 |
| Relation: |
https://www.frontiersin.org/articles/10.3389/fsens.2022.948466/full; https://doaj.org/toc/2673-5067 |
| DOI: |
10.3389/fsens.2022.948466 |
| Access URL: |
https://doaj.org/article/eafc6470e6b9457986314d517358fb58 |
| Accession Number: |
edsdoj.fc6470e6b9457986314d517358fb58 |
| Database: |
Directory of Open Access Journals |
