PDF(Pubmed)
Abstract：
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.
摘要：
空气传播的动物病毒病原体可以迅速传播并成为全球威胁，造成重大的社会经济和健康后果。预防和控制潜在的疫情暴发,准确,快,负担得起的即时护理（POC）测试至关重要。作为一个概念证明,我们已经开发了一种基于环介导等温扩增（LAMP）技术的分子系统，用于禽偏肺病毒（aMPV）检测，主要感染火鸡和鸡的空气传播剂。为此,通过将LAMP技术与特定的DNA官能化的AuNP（金纳米颗粒）偶联获得比色系统。使用从aMPV感染和未感染的鸡和火鸡收集的50个不同样品(咽拭子和气管组织)验证该系统。病毒检测可在约60分钟内用肉眼完成，对aMPV具有100%的特异性和87.88%的敏感性。总之,这种新颖的分子检测系统允许在现场进行合适的病毒检测，准确性和检测限（LOD）值高度接近基于qRT-PCR的诊断。此外，该系统可以轻松扩展到用于检测其他病毒的平台，解决当前在家禽养殖中用于病毒检测的POC测试可用性方面的差距。关键点:•使用RT-LAMP的aMPV诊断以高灵敏度和特异性实现。•使用DNA-纳米探针验证已经可视化了50个现场样品。•开发的系统是一个可靠的，快,和POCT的成本效益选择。
