Rapid Prototyping of Electrochemical DNA Detection Sensors
Bibliographic record
Abstract
Molecular diagnostic technologies can greatly improve the management of infectious diseases by offering reduced infection to detection window periods, rapid sample-to-answer times, and strain-specific pathogen identification1. In spite of great promise, these technologies have not made the predicted impact in resource poor regions with a high infectious disease burden. Handheld and chip-based nucleic acid detection systems are simple and inexpensive solutions that are envisioned to highly impact the diagnostic needs of these areas; however, their translation from the research lab to the marketplace has often been a lengthy process. To overcome this hurdle, we have developed a rapid prototyping method for fabricating integrated biosensing and sample preparation devices in a matter of hours. The fabrication method is focused on using multiple benchtop processes for creating hierarchical materials that are tunable in multiple lengthscales. Tunability in macroscale is achieved by using a CAD-driven craft cutter to create a shadow mask on a self-adhesive vinyl layer immobilized on a shrinkable polymer substrate. Gold thin films are deposited on the electrode by sputtering, and the gold-modified polymer substrate is heated in order to induce micro/nanostructuring on the gold substrate by shrinking the underlying substrate. For the purpose of DNA detection, these wrinkled gold electrodes are modified with a self-assembled monolayer of thiolated DNA probes, which is used for capturing specific DNA targets. In this work, we used the newly developed rapid prototyping method to create a multiplexed electrochemical DNA detection sensor. Through electrochemical measurements we demonstrated the surface area of these wrinkled electrodes to be six times larger than the surface area of planar electrodes of the same footprint. This allows more probe molecules to be immobilized on wrinkled electrodes compared to planar electrodes as indicated by fluorescence measurements. Furthermore, we combined these probe modified wrinkled electrodes with an electrocatalytic reporter system2, and successfully demonstrated that large signal changes (~100 % increase) are achieved when complementary targets are introduced, while negligible signal changes are observed in case of non-complementary targets. In addition, wrinkled electrodes transduce larger signal changes when compared to their planar electrode counterparts. In summary, we have developed a rapid prototyping method that is ideal for developing multi-scale electrodes for biosensing applications. In this work, we have demonstrated its application to electrochemical DNA sensing; however, this method can be used for creating systems that integrate sensing devices with sample preparation devices including bacterial lysis and magnetic separation 3components. (1) Gallarda, J.; Dragon, E. Blood Screening by Nucleic Acid Amplification Technology: Current Issues, Future Challenges. Mol. Diagnosis 2000, 5. (2) Lapierre, M. A.; Keefe, M. O.; Taft, B. J.; Kelley, S. O.; Merkert, E. F.; College, B.; Hill, C. Electrocatalytic Detection of Pathogenic DNA Sequences and Antibiotic Resistance Markers. 2003, 75, 6327–6333. (3) Hosseini, a.; Soleymani, L. Benchtop Fabrication of Multi-Scale Micro-Electromagnets for Capturing Magnetic Particles. Appl. Phys. Lett. 2014, 105, 074102. Figure 1
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How this classification was reachedexpand
Full frame machine prediction
Teacher imitationNot calibrated prevalence, not ground truth. Human validation pending. The Gemma side is a direct model label for every work in the frame, read from the title-only record. The Codex side is a classifier learned from the 10,348 direct Codex labels and calibrated to design-weighted sample rates; fields without enough sample support carry no Codex call. Candidate is the union of the two sides; consensus is their intersection. These outputs are machine_predicted_unvalidated and are not human labels.
Distilled classifier scores by category (both heads)
| Category | Codex | Gemma |
|---|---|---|
| Metaresearch | 0.001 | 0.002 |
| Meta-epidemiology (narrow) | 0.001 | 0.001 |
| Meta-epidemiology (broad) | 0.001 | 0.001 |
| Bibliometrics | 0.001 | 0.000 |
| Science and technology studies | 0.000 | 0.000 |
| Scholarly communication | 0.001 | 0.001 |
| Open science | 0.002 | 0.001 |
| Research integrity | 0.001 | 0.001 |
| Insufficient payload (model declined to judge) | 0.002 | 0.001 |
Machine scores (provisional)
The two teacher heads of the student model, read on this work. A score orders the frame for review; it never asserts a category, and the validation status ships verbatim with every row.
Baseline scores from an immature model (maturity gate not passed, 7 training rounds). Scores rank; they never assert a category.
score_only:v0-immature-baseline · verbatim from the scoring run: score_only means the number may rank works, and no category label ships from itClassification
machine, unvalidatedMachine predicted; a candidate call from one source (direct Gemma or distilled Codex), not a consensus.
How this classification was reached, model by model and score by score, is at the end of the page under "How this classification was reached".