Rapid detection of feline parvovirus using RAA-CRISPR/Cas12a-based lateral flow strip and fluorescence

文献类型： 外文期刊

第一作者： Chen, Han

作者： Chen, Han;Guo, Jie;Meng, Xiangshu;Yao, Mengfan;He, Longbin;Nie, Xiaoxuan;Xu, Han;Liu, Chao;Sun, Jian;Zhang, Jianlou;Wang, Jianke;Zhang, Hailing;Sun, Jian;Wang, Fei;Sun, Yuelong;Jiang, Zhong;He, Yanliang

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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

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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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