Development of a Gold Nanoparticle-Based Sensor for Authentication of Organic Milk Based on Differential Levels of miRNA
Abstract
:1. Introduction
2. Materials and Methods
2.1. Materials and Reagents
2.2. Synthesis of Gold Nanoparticles
2.3. Functionalization of AuNPs
2.4. Method Development
2.5. Sample Pre-Treatment
3. Results and Discussion
3.1. Method Development
3.1.1. Assay Optimization
Agg Probe-AuNP Ratio Optimization
MgCl2 Concentration, Hybridization Temperature, and Time Optimization
3.1.2. Analytical Characterization of the Method
3.2. Application of the Developed Sensor
4. Conclusions
Author Contributions
Funding
Data Availability Statement
Conflicts of Interest
References
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Conformation | Oligo Name | Sequence (5′→3′) |
---|---|---|
- | bta-miR-103 | AGC AGC AUU GUA CAG GGG UAU GA |
H-H | Agg/A m103 | Thiol-C6-AAA AAA AAA AAA CAA TGC TGC T |
Agg/B m103 | TCA TAG CCC TGT AAA AAA AAA-C3-Thiol | |
T-T | Agg/C m103 | Thiol-C6-AAA AAA AAA ATC ATA GCC CTG |
Agg/D m103 | TAC AAT GCT GCT AAA AAA AAA AA-C3-Thiol | |
- | bta-miR-155 | UUA AUG CUA AUC GUG AUA GGG GU |
H-H | Agg/A m155 | Thiol-C6-AAA AAA AAA AGA TTA GCA TTA A |
Agg/B m155 | ACC CCT ATC ACA AAA AAA AAA-C3-Thiol | |
T-T | Agg/C m155 | Thiol-C6-AAA AAA AAA ACC CCT ATC ACG |
Agg/D m155 | ATT AGC ATT AAA AAA AAA AAA-C3-Thiol | |
- | bta-miR-215 | AUG ACC UAU GAA UUG ACA GAC A |
H-H | Agg/A m215 | Thiol-C6-AAA AAA AAA ATC ATA GGT CAT |
Agg/B m215 | TGT CTG TCA ATA AAA AAA AAA-C3-Thiol | |
T-T | Agg/C m215 | Thiol-C6-AAA AAA AAA ATG TCT GTC AAT |
Agg/D m215 | TCA TAG GTC ATA AAA AAA AAA A-C3-Thiol |
Oligo Name | (Probe:AuNPs) Ratio |
---|---|
Agg/A m103 | 250:1 |
Agg/B m103 | 350:1 |
Agg/C m103 | 250:1 |
Agg/D m103 | 500:1 |
Agg/A m155 | 300:1 |
Agg/B m155 | 350:1 |
Agg/C m155 | 350:1 |
Agg/D m155 | 400:1 |
Agg/A m215 | 400:1 |
Agg/B m215 | 350:1 |
Agg/C m215 | 400:1 |
Agg/D m215 | 400:1 |
miRNA | Sample 1 (Ct) | Sample 2 (Ct) | Sample 3 (Ct) | Sample 4 (Ct) | ||||
---|---|---|---|---|---|---|---|---|
Original | Spiked | Original | Spiked | Original | Spiked | Original | Spiked | |
103 | 33.7 | 22 | 28.7 | 27.5 | 24.5 | 16 | 27.3 | 27.5 |
155 | 29.1 | 22 | 20.1 | 21.2 | 27.3 | 28.5 | 25.7 | 18.5 |
215 | 32.7 | 15.8 | 28.5 | 13.8 | 27.6 | 27.9 | 28.6 | 27.6 |
30a-5p | 30.9 | 27.4 | 23.7 | 22.9 | 22.7 | 25.5 | 23.5 | 22.7 |
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Lopez-Benitez, K.; Alcazar-Gonzalez, P.; el qassim, L.A.; Fernandez-Argüelles, M.T.; Vicente, F.; Royo, L.J.; Menendez-Miranda, M. Development of a Gold Nanoparticle-Based Sensor for Authentication of Organic Milk Based on Differential Levels of miRNA. Nanomaterials 2024, 14, 1364. https://doi.org/10.3390/nano14161364
Lopez-Benitez K, Alcazar-Gonzalez P, el qassim LA, Fernandez-Argüelles MT, Vicente F, Royo LJ, Menendez-Miranda M. Development of a Gold Nanoparticle-Based Sensor for Authentication of Organic Milk Based on Differential Levels of miRNA. Nanomaterials. 2024; 14(16):1364. https://doi.org/10.3390/nano14161364
Chicago/Turabian StyleLopez-Benitez, Karelmar, Patricia Alcazar-Gonzalez, Loubna Abou el qassim, Mª Teresa Fernandez-Argüelles, Fernando Vicente, Luis J. Royo, and Mario Menendez-Miranda. 2024. "Development of a Gold Nanoparticle-Based Sensor for Authentication of Organic Milk Based on Differential Levels of miRNA" Nanomaterials 14, no. 16: 1364. https://doi.org/10.3390/nano14161364