Design of Hydrogel Silk-Based Microarrays and Molecular Beacons for Reagentless Point-of-Care Diagnostics
We have developed a novel microarray system based on three technologies: 1) molecular beacons designed to interact with DNA targets at room temperature (25–27°C), 2) tridimensional silk-based microarrays containing the molecular beacons immersed in the silk hydrogel, and 3) shallow angle illuminatio...
| Main Authors: | , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Frontiers Media S.A.
2022-07-01
|
| Series: | Frontiers in Bioengineering and Biotechnology |
| Subjects: | |
| Online Access: | https://www.frontiersin.org/articles/10.3389/fbioe.2022.881679/full |
| _version_ | 1811222738213273600 |
|---|---|
| author | Alicia Sampieri Ricardo Monroy-Contreras Alexander Asanov Luis Vaca |
| author_facet | Alicia Sampieri Ricardo Monroy-Contreras Alexander Asanov Luis Vaca |
| author_sort | Alicia Sampieri |
| collection | DOAJ |
| description | We have developed a novel microarray system based on three technologies: 1) molecular beacons designed to interact with DNA targets at room temperature (25–27°C), 2) tridimensional silk-based microarrays containing the molecular beacons immersed in the silk hydrogel, and 3) shallow angle illumination, which uses separated optical pathways for excitation and emission. Unlike conventional microarrays that exhibit reduced signal-to-background ratio, require several stages of incubation, rinsing, and stringency control, and measure only end-point results, our microarray technology provides enhanced signal-to-background ratio (achieved by separating the optical pathways for excitation and emission, resulting in reduced stray light), performs analysis rapidly in one step without the need for labeling DNA targets, and measures the entire course of association kinetics between target DNA and the molecular beacons. To illustrate the benefits of our technology, we conducted microarray assays designed for the identification of influenza viruses. We show that in a single microarray slide, we can identify the virus subtype according to the molecular beacons designed for hemagglutinin (H1, H2, and H3) and neuraminidase (N1, N2). We also show the identification of human and swine influenza using sequence-specific molecular beacons. This microarray technology can be easily implemented for reagentless point-of-care diagnostics of several contagious diseases, including coronavirus variants responsible for the current pandemic. |
| first_indexed | 2024-04-12T08:20:03Z |
| format | Article |
| id | doaj.art-9fa2cbf8cbfc4373bd55d0a772a51147 |
| institution | Directory Open Access Journal |
| issn | 2296-4185 |
| language | English |
| last_indexed | 2024-04-12T08:20:03Z |
| publishDate | 2022-07-01 |
| publisher | Frontiers Media S.A. |
| record_format | Article |
| series | Frontiers in Bioengineering and Biotechnology |
| spelling | doaj.art-9fa2cbf8cbfc4373bd55d0a772a511472022-12-22T03:40:36ZengFrontiers Media S.A.Frontiers in Bioengineering and Biotechnology2296-41852022-07-011010.3389/fbioe.2022.881679881679Design of Hydrogel Silk-Based Microarrays and Molecular Beacons for Reagentless Point-of-Care DiagnosticsAlicia Sampieri0Ricardo Monroy-Contreras1Alexander Asanov2Luis Vaca3Departamento de Biología Celular y del Desarrollo, Instituto de Fisiología Celular, UNAM, Ciudad Universitaria, Mexico, MexicoDepartamento de Biología Celular y del Desarrollo, Instituto de Fisiología Celular, UNAM, Ciudad Universitaria, Mexico, MexicoTIRF Labs, Cary, NC, United StatesDepartamento de Biología Celular y del Desarrollo, Instituto de Fisiología Celular, UNAM, Ciudad Universitaria, Mexico, MexicoWe have developed a novel microarray system based on three technologies: 1) molecular beacons designed to interact with DNA targets at room temperature (25–27°C), 2) tridimensional silk-based microarrays containing the molecular beacons immersed in the silk hydrogel, and 3) shallow angle illumination, which uses separated optical pathways for excitation and emission. Unlike conventional microarrays that exhibit reduced signal-to-background ratio, require several stages of incubation, rinsing, and stringency control, and measure only end-point results, our microarray technology provides enhanced signal-to-background ratio (achieved by separating the optical pathways for excitation and emission, resulting in reduced stray light), performs analysis rapidly in one step without the need for labeling DNA targets, and measures the entire course of association kinetics between target DNA and the molecular beacons. To illustrate the benefits of our technology, we conducted microarray assays designed for the identification of influenza viruses. We show that in a single microarray slide, we can identify the virus subtype according to the molecular beacons designed for hemagglutinin (H1, H2, and H3) and neuraminidase (N1, N2). We also show the identification of human and swine influenza using sequence-specific molecular beacons. This microarray technology can be easily implemented for reagentless point-of-care diagnostics of several contagious diseases, including coronavirus variants responsible for the current pandemic.https://www.frontiersin.org/articles/10.3389/fbioe.2022.881679/fullmolecular beaconsmicroarrayssilkpoint-of-care diagnosticsinfluenza virus |
| spellingShingle | Alicia Sampieri Ricardo Monroy-Contreras Alexander Asanov Luis Vaca Design of Hydrogel Silk-Based Microarrays and Molecular Beacons for Reagentless Point-of-Care Diagnostics Frontiers in Bioengineering and Biotechnology molecular beacons microarrays silk point-of-care diagnostics influenza virus |
| title | Design of Hydrogel Silk-Based Microarrays and Molecular Beacons for Reagentless Point-of-Care Diagnostics |
| title_full | Design of Hydrogel Silk-Based Microarrays and Molecular Beacons for Reagentless Point-of-Care Diagnostics |
| title_fullStr | Design of Hydrogel Silk-Based Microarrays and Molecular Beacons for Reagentless Point-of-Care Diagnostics |
| title_full_unstemmed | Design of Hydrogel Silk-Based Microarrays and Molecular Beacons for Reagentless Point-of-Care Diagnostics |
| title_short | Design of Hydrogel Silk-Based Microarrays and Molecular Beacons for Reagentless Point-of-Care Diagnostics |
| title_sort | design of hydrogel silk based microarrays and molecular beacons for reagentless point of care diagnostics |
| topic | molecular beacons microarrays silk point-of-care diagnostics influenza virus |
| url | https://www.frontiersin.org/articles/10.3389/fbioe.2022.881679/full |
| work_keys_str_mv | AT aliciasampieri designofhydrogelsilkbasedmicroarraysandmolecularbeaconsforreagentlesspointofcarediagnostics AT ricardomonroycontreras designofhydrogelsilkbasedmicroarraysandmolecularbeaconsforreagentlesspointofcarediagnostics AT alexanderasanov designofhydrogelsilkbasedmicroarraysandmolecularbeaconsforreagentlesspointofcarediagnostics AT luisvaca designofhydrogelsilkbasedmicroarraysandmolecularbeaconsforreagentlesspointofcarediagnostics |