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从实验室到田间:用于早期检测短喙和侏儒综合征病毒核酸的创新型RPA-CRISPR/Cas12a平台

From lab to field: Innovative RPA‒CRISPR/Cas12a platform for early short-beak and dwarfism syndrome virus nucleic acids detection.

作者信息

Chen Xiuqin, Zhang Shizhong, Lin Su, Huang Meiqing, Chen Shilong, Wang Shao

机构信息

Institute of Animal Husbandry and Veterinary Medicine, Fujian Academy of Agricultural Science, Fuzhou, Fujian 350013, PR China; Fujian Animal Diseases Control Technology Development Center, Fuzhou, Fujian 350013, PR China.

Institute of Animal Husbandry and Veterinary Medicine, Fujian Academy of Agricultural Science, Fuzhou, Fujian 350013, PR China.

出版信息

Poult Sci. 2025 Apr 19;104(7):105191. doi: 10.1016/j.psj.2025.105191.

Abstract

Short beak and dwarfism syndrome virus (SBDSV) is the causative agent of short beak and dwarfism syndrome (SBDS), which is characterized by beak atrophy and dwarfism. SBDS has caused substantial economic losses for waterfowl husbandry industries. To address the urgent need for rapid and accurate SBDSV diagnostics, the study developed a dual-mode detection platform integrating recombinase-aided amplification (RPA) with CRISPR/Cas12a-mediated fluorescence and lateral flow strip readouts. The optimized assay achieved a detection limit of 10 copies/μL. Notably, the platform demonstrated superior specificity to distinguish SBDSV from genetically related Muscovy duck-origin goose parvovirus (GPV) and classical GPV, a distinction unachievable by qPCR. Clinical validation using 36 field samples confirmed 100% concordance with qPCR and indirect immunofluorescence assays, with no cross-reactivity against other common duck pathogens. This innovative detection system provides a robust toolkit for field-deployable SBDSV surveillance and lays a solid foundation for the development of novel diagnostic methodologies applicable to various waterfowl-related pathogens.

摘要

短喙侏儒症综合征病毒(SBDSV)是短喙侏儒症综合征(SBDS)的病原体,其特征为喙萎缩和侏儒症。SBDS给水禽养殖业造成了巨大经济损失。为满足快速准确诊断SBDSV的迫切需求,该研究开发了一种双模式检测平台,将重组酶辅助扩增(RPA)与CRISPR/Cas12a介导的荧光和侧流条读数相结合。优化后的检测方法实现了10拷贝/μL的检测限。值得注意的是,该平台在区分SBDSV与基因相关的番鸭源鹅细小病毒(GPV)和经典GPV方面表现出卓越的特异性,这是定量聚合酶链反应(qPCR)无法实现的区分。使用36份现场样本进行的临床验证证实,与qPCR和间接免疫荧光检测的一致性为100%,且与其他常见鸭病原体无交叉反应。这种创新的检测系统为可在现场部署的SBDSV监测提供了强大的工具包,并为开发适用于各种水禽相关病原体的新型诊断方法奠定了坚实基础。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9055/12051591/ca2599c2167f/gr1.jpg

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