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

小型企业创新研究(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率。这种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。这种由白斑综合症病毒(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。农民通常不会意识到疾病的问题，直到动物表现出疾病的晚期迹象或死亡。实验室诊断费用昂贵，结果需要几天到几周的时间，这往往太长了，结果无法付诸实施。对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，增加产业健壮性。该项目提出了一种新的诊断方法，在一种新的易于使用的现场测试中快速、容易和可靠地检测养殖对虾中的WSSV。这项SBIR第一阶段项目将推进一种新的WSSV诊断方法的转化，该技术将具有特定靶点的工程蛋白整合到纸基生物传感器中。以前的研究证明了纸基免疫分析的概念验证，聚合放大用于检测基于蛋白质的HRP2生物标记物-疟疾的主要蛋白质生物标记物-灵敏度低至70pM该项目将对耐热rcSso7d DNA结合蛋白进行生物工程，以结合WSSV靶标并将其整合到纸质测试格式中。将探索分析灵敏度、试剂组成和动态范围，作为了解所开发的分析的生物学和功能有意义的限度的一部分。任务包括一项WSSV挑战研究，以评估实验室养殖对虾样品的采样参数和检测下限。最终的原型将快速和选择性地绑定WSSV目标，并产生稳定、易于解释的比色结果。该奖项反映了NSF的法定使命，并通过使用基金会的智力优势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.