ORIGINAL RESEARCH article
Front. Microbiol.
Sec. Virology
This article is part of the Research TopicViral Diagnostics: Advancements for Rapid and Enhanced DetectionView all 5 articles
Rapid Visual Detection and Differentiation of Canine and Feline Parvovirus via a One-tube RPA-CRISPR/Cas13a Assay
Provisionally accepted
Zhurui Shao
Wenkai ZhangChangbin JinZhibo YangJingyi YangXiaohu ZhangXintong Huang
Yufei YangRuizi RenYiwen ZhangJieen Weng
Yueping Zhang*
Hao Shi*- China Agricultural University, Beijing, China
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Canine parvovirus type 2 (CPV-2) and feline parvovirus (FPV) are highly contagious pathogens that pose significant threats to domestic and wild carnivores. Rapid and accurate diagnosis is crucial for outbreak control and wildlife conservation, especially in resource-limited settings. In this study, we developed a one-tube RPA-CRISPR/Cas13a platform for the rapid detection and differentiation of CPV-2 and FPV. Two systems were established: a universal detection system for simultaneous identification of both viruses, and a differential detection system to distinguish between them. By targeting conserved regions of the VP2 gene and optimizing reaction conditions, we achieved high sensitivity and specificity. The universal system exhibited a limit of detection (LOD) of 102 copies/μL, while the differential system reached 103 copies/μL, with both assays completed within 60 minutes. Clinical validation using 50 samples showed 100% concordance with q-PCR and sequencing results. This study established a dual detection system that is sensitive, rapid, and suitable for use in primary-level settings and field conditions. It holds significant application value in enhancing the early diagnosis and differentiation of canine and feline parvoviruses, reducing the risk of transmission, and protecting the health of wildlife.
Keywords: CPV-2, CRISPR/Cas13a, detection, FPV, RPA, VP2 gene
Received: 30 Oct 2025; Accepted: 08 Dec 2025.
Copyright: © 2025 Shao, Zhang, Jin, Yang, Yang, Zhang, Huang, Yang, Ren, Zhang, Weng, Zhang and Shi. 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:
Yueping Zhang
Hao Shi
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