SBIR Phase I: Development of a field diagnostic for the rapid detection of White Spot Syndrome Virus (WSSV) in shrimp aquaculture

这个小企业创新研究（SBIR）第一阶段项目的更广泛影响/商业潜力是通过减少疾病影响来提高水产养殖生产力。疾病每年给水产养殖业造成数十亿美元的损失，并威胁到全球粮食供应。这种由白斑综合征病毒（WSSV）引起的疾病可以摧毁一个养虾场，每年造成超过10亿美元的损失。农民通常不知道疾病问题，直到动物出现疾病的晚期迹象或有死亡。实验室诊断是昂贵的，结果需要几天到几周的时间，这往往太长，结果无法采取行动。白斑病的早期预警可以使农民立即采取行动，减少损失，降低成本，提高工业稳健性。本项目提出了一种新的诊断方法，通过一种新的易于使用的现场测试，快速，轻松，可靠地检测养殖虾中的WSSV。这个SBIR一期项目将推进一种新的WSSV诊断技术的翻译，该技术将带有特定靶标的工程蛋白整合到基于纸张的生物传感器中。先前的研究证实了基于纸张的免疫分析的概念验证，该方法具有基于聚合扩增的方法，可用于检测基于蛋白质的hrp2生物标志物（疟疾的主要蛋白质生物标志物），灵敏度低至70pM。本项目将对耐热性rcSso7d dna结合蛋白进行生物工程改造，使其与WSSV靶点结合，并将其纳入纸质检测格式。分析灵敏度，试剂组成和动态范围将被探索作为努力的一部分，以了解开发的分析的生物学和功能上有意义的限制。任务包括WSSV挑战研究，以评估样品在实验室养殖虾中的采样参数和检出限。由此产生的原型将快速、选择性地结合WSSV靶标，并产生稳定、易于解释的比色结果。该奖项反映了美国国家科学基金会的法定使命，并通过使用基金会的知识价值和更广泛的影响审查标准进行评估，被认为值得支持。

The broader impact/commercial potential of this Small Business Innovation Research (SBIR) Phase I project is to increase aquaculture productivity by reducing disease impact. Disease costs billions in losses each year to the aquaculture industry and threatens the global food supply. The disease caused by White Spot Syndrome Virus (WSSV) can devastate a shrimp farm and costs over $1 billion in losses each year. Farmers are often unaware of a disease issue until animals show advanced signs of disease or there are mortalities. Laboratory diagnosis is expensive and results take days-to-weeks, which is often far too long for results to be actionable. Early warning of white spot disease can enable farmers to take immediate action to reduce losses, reducing costs and increasing industrial robustness. This project advances a new diagnostic to quickly, easily, and reliably detect WSSV in cultivated shrimp in a new easy-to-use, on-site test.This SBIR Phase I project will advance translation of a novel diagnostic for WSSV in a technology incorporating engineered proteins with specific targets into a paper-based biosensor. Prior studies demonstrated proof-of-concept with a paper-based immunoassay with polymerization-based amplification for detection of protein-based biomarkers for HRP2–the primary protein biomarker of malaria–with sensitivity as low as 70pM. This project will bioengineer thermostable rcSso7d DNA-binding proteins to bind the WSSV target and incorporate them into the paper testing format. Assay sensitivity, reagent composition, and dynamic range will be explored as part of an effort to understand the biological and functionally meaningful limits of the developed assay. Tasks include a WSSV challenge study to evaluate the sampling parameters and detection limits of the prototype in laboratory cultured shrimp. The resulting prototype will rapidly and selectively bind the WSSV target and yield stable, easily interpretable, colorimetric results.This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.