CRISPR-Cas12a/RPA Dual-Readout Assay for Rapid Field Detection of Porcine Rotavirus with Visualization
文献类型： 外文期刊
作者： Jiang, Xinjie 1 ; Huang, Yun 2 ; Jiang, Yi 1 ; Yang, Guang 3 ; Zheng, Xiaocong 2 ; Gao, Shuai 1 ;
作者机构： 1.Hainan Acad Agr Sci, Sanya Inst, Hainan Lab Anim Res Ctr, Sanya 572000, Peoples R China
2.Shenzhen Customs, Anim & Plant Inspect & Quarantine Technol Ctr, Shenzhen 518045, Peoples R China
3.China Agr Univ, Coll Anim Sci & Technol, Sanya Inst, Sanya 572025, Peoples R China
关键词： porcine rotavirus; CRISPR/Cas12a; RPA; visual detection; field-deployable diagnostics
期刊名称：VIRUSES-BASEL （ 影响因子：3.5； 五年影响因子：3.7 ）
ISSN：
年卷期： 2025 年 17 卷 7 期
页码：
收录情况： SCI
摘要： PoRV is a significant etiological agent of neonatal diarrhea in piglets, resulting in substantial economic losses within the global swine industry due to elevated mortality rates and reduced productivity. To address the urgent need for accessible and rapid diagnostics in resource-limited settings, we have developed a CRISPR/Cas12a-based assay integrated with recombinase polymerase amplification (RPA) for the visual detection of PoRV. This platform specifically targets the conserved VP6 gene using optimized RPA primers and crRNA, harnessing Cas12a's collateral cleavage activity to enable dual-readout via fluorescence or lateral flow dipsticks (LFDs). The assay demonstrates a detection limit of 102 copies/mu L within 1 h, exhibiting no cross-reactivity with phylogenetically related pathogens such as Transmissible Gastroenteritis Virus (TGEV). By eliminating reliance on thermal cyclers or specialized equipment, this method is fully deployable in swine farms, veterinary clinics, or field environments. The lateral flow format provides immediate colorimetric results that require minimal technical expertise, while the fluorescence mode allows for semi-quantitative analysis. This study presents a robust and cost-effective platform for decentralized PoRV surveillance in swine populations, addressing the critical need for portable diagnostics in resource-limited settings and enhancing veterinary health management.
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