Illegal, unreported and unregulated (IUU) fishing violates the rights of Indigenous peoples to traditional fishing grounds; compromises food security of legitimate fishers and coastal populations; and facilitates human labor trafficking. Once caught, tails, fins, and heads are cut off for storage and transportation, which makes identification by phenotype impossible. Polymerase Chain Reaction (PCR) and electrophoresis can detect different tuna species but are time-consuming and require access to a fully equipped laboratory and trained personnel. Thus, I developed a novel biological assay and a 3D printable portable device to detect Ahi tuna, one of the species of interest among three tuna species. The biological assay consists of three steps: DNA extraction by crude lysis method; DNA amplification by Recombinase Polymerase Amplification (RPA); and detection by fluorescence using Clustered Regularly Interspaced Short Palindromic Repeats associated (CRISPR Cas) 12a. qPCR verified Ahi tuna DNA extraction by crude lysis method, generating 10⁶ copies of DNA per reaction comparable to the standard silica-based capture method. Nested qPCR and Tapestation verified RPA’s successful amplification of Ahi tuna DNA extracted by crude lysis method. Quantification of fluorescence by qPCR verified that one-pot RPA and CRISPR Cas12a reactions could generate up to 6000 Relative Fluorescence Units above the negative control within 15 minutes. The isothermal device keeps the samples at a constant temperature (37-42℃) for the RPA and CRISPR Cas12a reactions. I successfully demonstrated a <50 min sample-to-result assay to detect Ahi tuna DNA, and this protocol will be further adapted for testing other types of tuna including Bluefin tuna. By developing such a rapid and affordable isothermal biological assay and a point-of-need 3D printable portable device for tuna identification, I aim to contribute in helping individuals and ecosystems impacted by IUU fishing.

In this research project, our multidisciplinary team is developing environmental forensic technology to identify illegally caught seafood, with a focus on high-value species such as bluefin tuna. To tackle the difficult task of immediately detecting illegal, unregulated, and unreported (IUU) fishing, we are developing a rapid, affordable, and portable detection device that uses isothermal amplification and blue LEDs to detect resultant green fluorescent products indicating the presence of tuna DNA. As the lead Electrical Engineer, I designed an electrical circuit to precisely and stably control the temperature of a resistive heater using a microcontroller, thermistor, and PID algorithm along with an LED circuit. The requirement for the process involves the utilization of isothermal amplification technique at a fixed temperature of 37°C (33-42°C range) and green fluorescence detection (at 550 nm) in DNA using blue LEDs (at 470 nm) and an orange acrylic filter to filter the blue wavelength. The result is a circuit that meets the requirements for the biochemical process and enables real-time feedback without the need for shipping samples to a laboratory. With the development of the device finished, the next step is to streamline the user experience. I am leading the software development effort to create a phone application that facilitates the assay setup process and automates image capturing and analysis. With a capable research team, including colleagues shaping the user interface, a junior researcher connecting the device to the phone application, and a Ph.D. candidate in Computer Science and Engineering developing a proprietary camera software, our team's effort culminates in a user-friendly phone app that streamlines the assay workflow and provides real-time sample analysis. With the completion of this project, we will be able to make a difference by helping communities and marine ecosystems.