ORIGINAL RESEARCH article
Front. Microbiol.
Sec. Virology
Volume 16 - 2025 | doi: 10.3389/fmicb.2025.1617955
This article is part of the Research TopicViral Diagnostics: Advancements for Rapid and Enhanced DetectionView all articles
Rapid, sensitive, and visual detection of Porcine Rotavirus with RPA-CRISPR/Cas13
Provisionally accepted
- College of Veterinary Medicine, Sichuan Agricultural University, Chengdu, China
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Porcine rotavirus (PoRV) is one of the major pathogens causing viral enteritis in piglets, posing serious threats to the pig industry and public health. Existing pathogen detection methods, such as RT-qPCR, suffer from complex procedures and strong reliance on equipment, making them difficult to meet the needs of grassroots laboratories or field detection. Therefore, in this study, a novel rapid and visual detection platform, was developed based on the CRISPR/Cas13 system. This platform integrates Recombinase Polymerase Amplification (RPA), T7 transcription, CRISPR trans-cleavage, and fluorescence signal output. Targeting the PoRV VP6 gene, it achieves highly sensitive and specific detection of PoRV without the need for large equipment. The optimized system has a minimum detection limit of 10¹ copies/μL for standard plasmids, and it was validated with 62 clinical diarrheal samples, yielding results consistent with RT-qPCR, demonstrating good clinical applicability. This platform is simple to operate, intuitive in detection, and cost-effective, making it suitable for rapid field screening of PoRV in resource-limited areas. It provides robust technical support for early diagnosis and epidemiological investigation of porcine rotavirus.
Keywords: Porcine rotavirus, RPA, CRISPR/Cas13, on-site detection, One-tube
Received: 25 Apr 2025; Accepted: 17 Jul 2025.
Copyright: © 2025 Mi, Dai, Xing, Zhang, Xu and Zhu. This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) or licensor are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
* Correspondence:
Zhiwen Xu, College of Veterinary Medicine, Sichuan Agricultural University, Chengdu, China
Ling Zhu, College of Veterinary Medicine, Sichuan Agricultural University, Chengdu, China
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