用于猪流行性腹泻病毒即时检测的纸基微流控芯片的研制

Development of a Paper-Based Microfluidic Chip for Point-of-Care Detection of PEDV.

作者信息

Li Renfeng, Tian Xiangqin, Cao Wenyan, Jiang Jiaxin, Yuan Jiakang, Li Linyue, You Yonghe, Zhou Yanlin, Wang Ziliang, Wang Fangyu

机构信息

College of Animal Science and Veterinary Medicine, Henan Institute of Science and Technology, Xinxiang 453003, China.

Henan Key Laboratory of Medical Tissue Regeneration, Xinxiang Medical University, Xinxiang 453003, China.

出版信息

Vet Sci. 2025 Apr 30;12(5):427. doi: 10.3390/vetsci12050427.

Abstract

PEDV poses a significant threat to the global swine industry, necessitating rapid and accurate diagnostic methods for effective disease management. In this study, we developed a foldable, easy-to-use paper-based microfluidic analytical device (PAD) for on-site detection of PEDV. The device seamlessly integrates paper-based nucleic acid enrichment, LAMP reaction, and visual lateral flow detection into a single platform. Key parameters, including nucleic acid extraction protocols, chromatographic channel configurations, colorimetric indicators, and reaction temperature and duration, were systematically optimized. The resulting LAMP-PAD assay detects PEDV within 30 min at 60 °C, achieving a limit of detection of 4.82 × 10 copies/μL with no cross-reactivity against other viruses. When evaluated against RT-PCR using clinical specimens, the assay demonstrated a specificity of 100%, a sensitivity of 95.3%, and an overall concordance of 98.5%. This paper-based sensor offers a promising alternative for the rapid, on-site detection of PEDV and other highly transmissible pathogens.

摘要

猪流行性腹泻病毒(PEDV)对全球养猪业构成重大威胁,因此需要快速准确的诊断方法来进行有效的疾病管理。在本研究中,我们开发了一种可折叠、易于使用的纸质微流控分析装置(PAD),用于现场检测PEDV。该装置将基于纸的核酸富集、环介导等温扩增(LAMP)反应和可视化侧向流动检测无缝集成到一个平台中。对包括核酸提取方案、色谱通道配置、比色指示剂以及反应温度和持续时间在内的关键参数进行了系统优化。所得的LAMP-PAD检测法在60℃下30分钟内即可检测到PEDV,检测限达到4.82×10拷贝/μL,且与其他病毒无交叉反应。当使用临床样本与逆转录聚合酶链反应(RT-PCR)进行评估时,该检测法的特异性为100%,灵敏度为95.3%,总体一致性为98.5%。这种基于纸的传感器为快速现场检测PEDV和其他高传播性病原体提供了一种有前景的替代方法。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0535/12115601/c153597d7294/vetsci-12-00427-g001.jpg

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