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基于金磁纳米颗粒的表面增强拉曼散射免疫层析法对猪流行性腹泻病毒和猪轮状病毒共感染的快速现场诊断

Rapid on-site diagnosis of PEDV and PoRV co-infection by gold magnetic nanoparticles-based SERS immunochromatography.

作者信息

He Zhaoyuan, Duan Xiaoge, Zhao Zhi, Chen Yingkai, Fu Chengxiang, Zhang Fuxin, Wang Jinzi, Feng Jianyuan, Lin Nanxin, Chen Hailan

机构信息

College of Animal Science and Technology, Guangxi Key Laboratory of Animal Breeding, Disease Control and Prevention, Guangxi University, Nanning, 530004, China.

School of Marine Sciences and Biotechnology, Guangxi Minzu University, Nanning, 530006, Guangxi, China.

出版信息

Talanta. 2025 Apr 1;285:127428. doi: 10.1016/j.talanta.2024.127428. Epub 2024 Dec 19.

Abstract

Porcine epidemic diarrhea virus (PEDV) and porcine rotavirus (PoRV) are the two main pathogens causing porcine diarrhea, which are characterized by high morbidity and mortality. Most of the diagnostic methods available are limited to the laboratory or fail to highlight their advantages in terms of target species, detection time, sensitivity, and stability. To meet the demand for rapid on-site diagnosis of PEDV and PoRV co-infection, a surface-enhanced Raman scattering (SERS) immunochromatographic sensor based on gold magnetic nanoparticles (MNPs) was developed. The sensor is dual-mode, detecting on the basis of color signals by the naked eye and Raman signals. After a series of optimizations, the constructed sensor could perform simultaneous qualitative and quantitative detection of PEDV and PoRV in just 18 min, with visualized (color signals observed by the naked eye) limits of detection (LOD) of 3.13 × 10 TCID/mL and 4.69 × 10 copies/μL, respectively. The LOD based on Raman signal analysis was as low as 4.63 × 10 TCID/mL and 3.30 × 10 copies/μL for PEDV and PoRV, respectively. In addition, the sensor exhibited excellent specificity without cross-reactivity with common pathogens. The overall compliance rate with RT-PCR was 92.1 % (35/38) for 38 clinical samples. Therefore, the sensor is characterized by high sensitivity, specificity, reproducibility, and accuracy, making it suitable for the simultaneous rapid on-site detection of PEDV and PoRV.

摘要

猪流行性腹泻病毒(PEDV)和猪轮状病毒(PoRV)是引起猪腹泻的两种主要病原体,其特点是发病率和死亡率高。现有的大多数诊断方法仅限于实验室使用,或者在目标物种、检测时间、灵敏度和稳定性方面未能突出其优势。为了满足对PEDV和PoRV混合感染进行快速现场诊断的需求,开发了一种基于金磁纳米颗粒(MNPs)的表面增强拉曼散射(SERS)免疫层析传感器。该传感器具有双模式,可基于肉眼可见的颜色信号和拉曼信号进行检测。经过一系列优化后,构建的传感器能够在短短18分钟内同时对PEDV和PoRV进行定性和定量检测,可视化(肉眼观察颜色信号)检测限(LOD)分别为3.13×10 TCID/mL和4.69×10拷贝/μL。基于拉曼信号分析的PEDV和PoRV的LOD分别低至4.63×10 TCID/mL和3.30×10拷贝/μL。此外,该传感器表现出优异的特异性,与常见病原体无交叉反应。对于38份临床样本,与RT-PCR的总体符合率为92.1%(35/38)。因此,该传感器具有高灵敏度、特异性、重现性和准确性的特点,适用于PEDV和PoRV的同时快速现场检测。

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