College of Veterinary Medicine, Institute of Comparative Medicine, Yangzhou University, Yangzhou 225009, China.
Jiangsu Co-Innovation Center for Prevention and Control of Important Animal Infectious Diseases and Zoonoses, Yangzhou University, Yangzhou 225009, China.
Biosensors (Basel). 2022 Dec 8;12(12):1146. doi: 10.3390/bios12121146.
Porcine epidemic diarrhea virus (PEDV), a coronavirus that causes highly infectious intestinal diarrhea in piglets, has led to severe economic losses worldwide. Rapid diagnosis and timely supervision are significant in the prophylaxis of PEDV. Herein, we proposed a gold-nanorod (GNR) probe-assisted counting method using dark field microscopy (DFM). The antibody-functionalized silicon chips were prepared to capture PEDV to form sandwich structures with GNR probes for imaging under DFM. Results show that our DFM-based assay for PEDV has a sensitivity of 23.80 copies/μL for simulated real samples, which is very close to that of qPCR in this study. This method of GNR probes combined with DFM for quantitative detection of PEDV not only has strong specificity, good repeatability, and a low detection limit, but it also can be implemented for rapid on-site detection of the pathogens.
猪流行性腹泻病毒（PEDV）是一种冠状病毒，可导致仔猪高度传染性肠道腹泻，已在全球范围内造成严重的经济损失。快速诊断和及时监测对于预防 PEDV 至关重要。在此，我们提出了一种基于金纳米棒（GNR）探针的计数方法，该方法使用暗场显微镜（DFM）进行检测。制备了抗体功能化的硅片以捕获 PEDV，然后与 GNR 探针形成夹心结构，用于 DFM 下成像。结果表明，我们基于 DFM 的 PEDV 检测方法对模拟实际样本的灵敏度为 23.80 拷贝/μL，与本研究中的 qPCR 非常接近。该方法结合了 GNR 探针和 DFM 用于定量检测 PEDV，不仅具有很强的特异性、良好的重复性和较低的检测限，而且还可以实现对病原体的快速现场检测。
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