BioAssays SL. Parque Científico de Madrid, Madrid, Spain.
Research Group of "Animal Viruses" of Complutense University of Madrid, Madrid, Spain.
Appl Microbiol Biotechnol. 2024 Jul 10;108(1):414. doi: 10.1007/s00253-024-13243-x.
Airborne animal viral pathogens can rapidly spread and become a global threat, resulting in substantial socioeconomic and health consequences. To prevent and control potential epidemic outbreaks, accurate, fast, and affordable point-of-care (POC) tests are essential. As a proof-of-concept, we have developed a molecular system based on the loop-mediated isothermal amplification (LAMP) technique for avian metapneumovirus (aMPV) detection, an airborne communicable agent mainly infecting turkeys and chickens. For this purpose, a colorimetric system was obtained by coupling the LAMP technique with specific DNA-functionalized AuNPs (gold nanoparticles). The system was validated using 50 different samples (pharyngeal swabs and tracheal tissue) collected from aMPV-infected and non-infected chickens and turkeys. Viral detection can be achieved in about 60 min with the naked eye, with 100% specificity and 87.88% sensitivity for aMPV. In summary, this novel molecular detection system allows suitable virus testing in the field, with accuracy and limit of detection (LOD) values highly close to qRT-PCR-based diagnosis. Furthermore, this system can be easily scalable to a platform for the detection of other viruses, addressing the current gap in the availability of POC tests for viral detection in poultry farming. KEY POINTS: •aMPV diagnosis using RT-LAMP is achieved with high sensitivity and specificity. •Fifty field samples have been visualized using DNA-nanoprobe validation. •The developed system is a reliable, fast, and cost-effective option for POCT.
空气传播的动物病毒病原体传播迅速，可能成为全球性威胁，给社会经济和健康带来重大影响。为了预防和控制潜在的疫情爆发，准确、快速且经济实惠的即时检测（POC）至关重要。作为概念验证，我们开发了一种基于环介导等温扩增（LAMP）技术的分子系统，用于检测禽偏肺病毒（aMPV），这是一种主要感染火鸡和鸡的空气传播传染性病原体。为此，我们通过将 LAMP 技术与特定的 DNA 功能化金纳米颗粒（AuNPs）偶联，获得了比色系统。该系统使用 50 个来自感染和未感染的鸡和火鸡的咽拭子和气管组织的不同样本进行了验证。用肉眼观察，病毒检测可在大约 60 分钟内完成，对 aMPV 的特异性为 100%，敏感性为 87.88%。总之，这种新型分子检测系统可用于现场进行适当的病毒检测，其准确性和检测限（LOD）值与基于 qRT-PCR 的诊断非常接近。此外，该系统可以很容易地扩展到用于检测家禽养殖中其他病毒的平台，解决了目前在 POC 检测方面缺乏病毒检测的问题。关键点： • 使用 RT-LAMP 进行 aMPV 诊断具有高灵敏度和特异性。 • 通过 DNA-纳米探针验证可视化了 50 个现场样本。 • 开发的系统是一种可靠、快速且具有成本效益的 POCT 选择。
