Rapid detection of feline parvovirus using RAA-CRISPR/Cas12a-based lateral flow strip and fluorescence
文献类型： 外文期刊
作者： Chen, Han 1 ; Zhang, Hailing 2 ; Guo, Jie 1 ; Meng, Xiangshu 1 ; Yao, Mengfan 1 ; He, Longbin 1 ; Nie, Xiaoxuan 1 ; Xu, Han 1 ; Liu, Chao 1 ; Sun, Jian 1 ; Wang, Fei 4 ; Sun, Yuelong 4 ; Jiang, Zhong 5 ; He, Yanliang 6 ; Zhang, Jianlou 1 ; Wang, Jianke 1 ;
作者机构： 1.Hebei Agr Univ, Coll Vet Med, Hebei Vet Biotechnol Innovat Ctr, Baoding, Peoples R China
2.Chinese Acad Agr Sci, Inst Special Anim & Plant Sci, Key Lab Special Anim Epidem Dis, Minist Agr, Changchun, Peoples R China
3.Weihai Ocean Vocat Coll, Weihai, Peoples R China
4.Chongqing Three Gorges Vocat Coll, Chongqing, Peoples R China
5.Agr Bureau Zhuozhou City, Zhuozhou, Peoples R China
6.Sinovet Jiangsu Biopharmaceut Co Ltd, Taizhou, Peoples R China
关键词： CRISPR/Cas12a; detection; feline; parvovirus; RAA; lateral flow strip
期刊名称：FRONTIERS IN MICROBIOLOGY （ 影响因子：4.5； 五年影响因子：5.2 ）
ISSN：
年卷期： 2025 年 16 卷
页码：
收录情况： SCI
摘要： 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 x 100 copies of recombinant plasmids per reaction, compared with 2.1 x 103 copies for conventional PCR analysis. Furthermore, no cross-reactivity was observed for the RAA-CRISPR/Cas12a assay with feline coronavirus, feline herpesvirus, and feline calicivirus, demonstrating reasonable specificity. Additionally, 43 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.
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