Cea-Callejo Pablo, Trenado Claudia, El Mansouri Elías, Gomez-Lucia Esperanza, Doménech Ana, Biarnés Mar, Cuenca J Marco, Sánchez-Llatas Christian J, Madrid Ricardo, Benítez Laura
Department of Genetics, Physiology, and Microbiology, School of Biology, Complutense University of Madrid (UCM), 28040 Madrid, Spain.
Research Group of "Animal Viruses", Complutense University of Madrid, 28040 Madrid, Spain.
Int J Mol Sci. 2025 Feb 25;26(5):1971. doi: 10.3390/ijms26051971.
Infectious laryngotracheitis virus (ILTV), a DNA virus classified as , causes a highly contagious respiratory disease in chickens, leading to significant economic losses and health risks for the poultry industry. The rapid detection of ILTV is essential to control its spread and prevent outbreaks. Traditional diagnostic methods like PCR are costly, require specialized personnel, and delay response efforts. To address this, we developed a point-of-care diagnostic test combining loop-mediated isothermal amplification (LAMP) with DNA nanoprobes on respiratory swabs. LAMP targets the ILTV-glycoprotein E (gE) gene, enabling rapid nucleic acid amplification at 65 °C without extraction, making it suitable for on-site detection. DNA nanoprobes provide a colorimetric readout visible to the naked eye. Gold nanoparticles drive this readout, as their red color, based on localized surface plasmon resonance, persists in the presence of ILTV DNA through DNA-DNA hybridization, ensuring reliable detection. The assay achieved 100% sensitivity and specificity for ILTV-gE, with a detection limit of 200 copies per reaction, allowing for the early identification of infections. The results are available within 45 min, enabling prompt measures to control ILTV spread. Cost-effective and user-friendly, this method enhances disease management and biosecurity in poultry farms.
传染性喉气管炎病毒（ILTV）是一种归类为 的DNA病毒，可在鸡群中引发高度传染性呼吸道疾病，给家禽业带来重大经济损失和健康风险。快速检测ILTV对于控制其传播和预防疫情爆发至关重要。像PCR这样的传统诊断方法成本高昂，需要专业人员操作，且会延误应对工作。为解决这一问题，我们开发了一种即时诊断测试，将环介导等温扩增（LAMP）与用于呼吸道拭子的DNA纳米探针相结合。LAMP靶向ILTV糖蛋白E（gE）基因，能够在65°C下无需提取即可快速进行核酸扩增，适用于现场检测。DNA纳米探针提供肉眼可见的比色读数。金纳米颗粒驱动这一读数，因为基于局域表面等离子体共振，其红色在存在ILTV DNA时通过DNA-DNA杂交得以保留，确保可靠检测。该检测方法对ILTV-gE的灵敏度和特异性均达到100%，检测限为每个反应200个拷贝，能够早期识别感染情况。结果可在45分钟内获得，从而能够迅速采取措施控制ILTV传播。这种方法经济高效且用户友好，可加强家禽养殖场的疾病管理和生物安全。
