◻ Development of a bivalent vaccine for effective protection against red sea bream iridoviral disease (RSIVD)

 • Predicting and comparing the antigenic determinants of candidate vaccine strains by targeting antigen-related genes to select strains capable of inducing strong immune responses.

 • MCP plays an important role in virus attachment and penetration, and is known as a key antigen site in vaccine development for iridovirus due to its high antigenicity.

◻ Development of an oral and immersion vaccine system based on natural polymers 

 • The largest proportion of commercially available vaccines are injectable formulations. However, none are targeted at ornamental fish, and injectable formulations used in aquaculture are difficult to apply to smaller ornamental fish.

 • Based on natural polymers with the advantages of biodegradability and non-toxicity, research is being conducted on the development of oral and immersion vaccines for viral diseases by utilizing nanotechnology with a high absorption rate. 

◻ Development of In silico-based epitope vaccine system

 • Fish vaccines have traditionally relied on inactivated vaccines due to safety concerns; however, this approach is ineffective in rapidly responding to highly mutagenic viruses.

 • Epitope-based vaccine design and efficacy assessment through in silico analysis can significantly reduce the time required for vaccine development and enhance efficiency, thereby laying the foundation for a more effective vaccination strategy in aquaculture.

◻ Study evaluating various antiviral drugs against fish virus

• Viral diseases affect numerous fish species every year, leading to significant damage. However, due to the lack of effective treatment and prevention measures for fish viruses, it is crucial to develop strategies for controlling and treating these diseases. To address this, we explored various substances and confirmed that they not only reduced the viral titer in vitro but also improved the survival rates in fish. This research will enable the preparation of rapid response plans, and we are currently developing an antiviral screening tool using genetic engineering technology.

◻ Establishing aquatic animal disease scenarios based on epidemiological modeling

 • Disease risk factors refer to any biological or environmental factors that increase the probability of a specific disease outbreak in an individual or population.

 • A disease forecasting model is a system for predicting diseases based on past incidence and epidemiological data, as well  as risk-associated factors of diseases.

◻ Development of eDNA(environmental DNA) diagnostic method

 • Development and application of environmental DNA tools

◻ Development of a rapid molecular diagnostic method combining POCT-based LFA with isothermal amplification

 • Molecular diagnostic methods, such as PCR, are widely studied for virus detection. While these methods offer high specificity and sensitivity, they have drawbacks, such as long analysis times and the need for sophisticated equipment. Consequently, applying them for rapid diagnostics in non-specialized laboratories or in the field can be challenging. Therefore, there is a growing need for the development of faster and simpler diagnostic methods.

◻ Development of Recombinase polymerase amplification (RPA) diagnostic method 

• Recombinase polymerase amplification (RPA) is a technique for rapidly amplifying targeted DNA fragments in as little as 30 minutes at a constant temperature of 25 °C to 42 °C, using three main enzymes: recombinase, single-strand binding protein (SSB), and strand substitution polymerase.

• Recombinase polymerase amplification (RPA) is a highly suitable isothermal amplification assay for multiplexing because, like PCR, it uses two opposing primers, making it more advantageous compared to other isothermal nucleic acid amplification techniques such as LAMP (Loop-mediated Isothermal Amplification) and CPA (Cross Priming Amplification).

• LAMP and CPA each require 4 to 5 primers, whereas RPA uses only two primers, which reduces complexity and minimizes the risk of non-specific amplification. This makes RPA more suitable for detecting multiple targets simultaneously in a single reaction.

◻ Development of viability-quantitative PCR with propidium monoazide for selective detection of white spot syndrome virus

• Ordinary PCR and quantitative PCR (qPCR) amplify the DNA of viruses without regard to the integrity of capsid and envelope that is an important criterion for viability and infectivty of virus.

• Viability dye like propidium monoazide used in viability-quantitative PCR (v-qPCR) selectively penetrate the compromised capsid and envelope. Upon reaction with propidium monoazide, the DNA is cross-linked with the azide group of propidium monoazide by exposure to the light. This photolysis causes a structural change in nucleic acid to inhibit elongation by polymerase and renders the DNA insoluble in water to be lost during DNA extraction.

• viability-quantitative PCR (v-qPCR) v-qPCR is an efficient and rapid diagnostic method to assess infectivity of viruses in the absence of a reliable cell culture method through selective detection.