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ORIGINAL RESEARCH article
Front. Microbiol.
Sec. Virology
Volume 16 - 2025 | doi: 10.3389/fmicb.2025.1501635
Rapid Detection of Feline Parvovirus Using RAA-CRISPR/Cas12a-Based Lateral Flow Strip and Fluorescence
Provisionally accepted
Han Chen 1
Hailing Zhang 2 Jie Guo 3 Xiangshu Meng 3 Mengfan Yao 3 Longbin He 3 Xiaoxuan Nie 3 Han Xu 3 Chao Liu 3 Jian Sun 3 Fei Wang 3 Yuelong Sun 4 Zhong Jiang 5
He Yanliang 6
Jianke Wang 1*- 1 School of Animal Medicine, Hebei Agricultural University, Baoding, China
- 2 Institute of Special Animal and Plant Sciences, Chinese Academy of Agricultural Sciences, Changchun, Jilin Province, China
- 3 College of Veterinary Medicine, Hebei Agricultural University, Baoding, China
- 4 New Ruipeng Pet Group Inc (Beijing) Co., Ltd., Beijing, China
- 5 Agriculture Bureau of Zhuozhou City, Zhuozhou, China
- 6 Sinovet (Jiangsu) Biopharmaceuticals Co., Ltd., Taizhou, China
Feline parvovirus (FPV) causes severe gastroenteritis and leukopenia in cats, with high morbidity and mortality, necessitating a rapid and effective antigen diagnostic test with high sensitivity and specificity. In this study, a diagnostic platform based on a combination of Recombinase-Aided Amplification (RAA) and CRISPR/Cas12a was established for detecting FPV. Cas12a recombinant protein was purified using Nickel-Nitriloacetic Acid resin after heterologous expression in Escherichia coli. The results of RAA-CRISPR/Cas12a can be detected with a fluorescence reader or lateral flow strips (LFS) for on-site detection. The RAA-CRISPR/Cas12a-LFS had a detection limit of 2.1×10 0 copies of recombinant plasmids per reaction, compared with 2.1×10 3 copies for conventional PCR analysis. Furthermore, no crossreactivity was observed for the RAA-CRISPR/Cas12a assay with feline coronavirus, feline herpesvirus, and feline calicivirus, demonstrating reasonable specificity. Additionally, fortythree cat fecal samples with suspected clinical signs were assayed with RAA-CRISPR/Cas12a-LFS and conventional PCR in parallel. The RAA-CRISPR/Cas12a-LFS showed a 100% coincident rate with PCR. In summary, a novel, visual, sensitive, and specific detection assay based on RAA and CRISPR/Cas12a was developed for FPV.
Keywords: CRISPR/Cas12a, detection, feline, Parvovirus, RAA, Lateral flow strip
Received: 25 Sep 2024; Accepted: 13 Feb 2025.
Copyright: © 2025 Chen, Zhang, Guo, Meng, Yao, He, Nie, Xu, Liu, Sun, Wang, Sun, Jiang, Yanliang and Wang. 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:
Jianke Wang, School of Animal Medicine, Hebei Agricultural University, Baoding, China
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