Developing a rapid, adaptable point-of-care diagnostic pipeline and device for the fast-mutating PRRSV

猪繁殖与呼吸综合征（PRRS），或称蓝耳病，给全球养猪业造成了相当大的经济损失。该病毒的高突变率导致新变种的出现和成年猪的再感染，使当前的控制措施具有挑战性。疫苗对新毒株提供的保护有限，因此迫切需要制定有效的控制规划，以减轻病毒的影响。然而，市售的用于检测病毒的聚合酶链反应试剂盒灵敏度有限，而且每年只更新一次，因此不足以检测新出现的毒株。迫切需要一种能够快速准确地检测出所有PRRS菌株的新的诊断检测方法。我们提出的解决方案利用高通量测序、机器学习辅助引物设计、快速响应验证和定制的护理点检测（POCT）来彻底改变PRRS监测。这种方法提高了控制措施的效率，并能够适应当地出现的快速变异病毒株，如PRRS。我们现有的基于环介导等温扩增技术的POCT设备和检测试剂盒对非洲猪瘟病毒具有较高的敏感性和特异性。我们计划修改我们目前的POCT产品，设计一个PRRS面板，以检测西班牙流行菌株并进行基因分型。我们将与我们的西班牙合作伙伴合作，从我们定期从猪场收集的样本中生成PRRSV全基因组序列数据库，使我们能够快速应对新出现的菌株。实施快速和可靠的PRRS诊断系统将在猪的家畜管理方面带来前所未有的效率和可靠性。这反过来又会提高农业生产率，从而促进经济发展。此外，我们的技术将与消费者价值观保持一致，促进道德和可持续的农业实践，并显著改善动物福利。促进生猪养殖的社会影响是我们创新生物技术研究的重要成果。

Porcine reproductive and respiratory syndrome (PRRS), or blue ear disease, causes considerable economic losses in the global pig farming industry. The virus's high mutation rate leads to the emergence of new variants and reinfections in adult pigs, making current control measures challenging. Vaccines provide limited protection against new strains, underscoring the critical need for effective control programs to mitigate the virus's impact.However, commercially available PCR kits used to detect the virus have limited sensitivity and are update only once a year, making them inadequate for detecting emerging strains. A new diagnostic test that can quickly and accurately detect all PRRS strains is urgently required.Our proposed solution utilizes high-throughput sequencing, machine-learning-aided primer design, fast response validation, and customized point-of-care testing (POCT) to revolutionize PRRS surveillance. This approach improves control measures' efficiency and enables adaptation to locally emerging strains of fast-mutating viruses like PRRS.Our existing POCT device and testing kit based on loop-mediated isothermal amplification technology have demonstrated high sensitivity and specificity against the African swine fever virus. We plan to modify our current POCT products to design a PRRS panel to detect and genotype prevalent strains in Spain. We will build a database of whole-genome PRRSV sequences generated from samples we regularly collect from pig farms in collaboration with our Spanish partners, enabling us to respond to emerging strains quickly.Implementing fast and reliable diagnosis systems against PRRS will lead to unprecedented efficiency and reliability in managing pig livestock. This, in turn, will improve agricultural productivity, resulting in economic progress. Moreover, our technology will align with consumer values, fostering ethical and sustainable farming practices and significantly improving animal welfare. Advancing the social impact of pig farming is an essential outcome of our innovative biotechnology research.