Li Hao, Wang Guochun, Wu Xin, Ying Xinyu, Luo Zhidan, Lu Chen, Zhang Ping
College of Life Sciences, Hunan Normal University, Changsha, China.
Jiangsu Key Laboratory of Marine Biological Resources and Environment, Jiangsu Key Laboratory of Marine Pharmaceutical Compound Screening, Jiangsu Ocean University, Lianyungang, China.
Curr Microbiol. 2025 Aug 13;82(10):450. doi: 10.1007/s00284-025-04375-y.
Amanita fuliginea is a common lethal mushroom with high mortality rates due to its ease of ingestion and concealed early symptoms. Rapid and specific identification of this species is crucial for clinical diagnosis and treatment. However, the existing rapid detection methods for nucleic acids of A. fuliginea were still plagued by specificity issues. Herein, we developed a recombinase polymerase amplification (RPA) coupled with CRISPR/Cas12b and lateral flow strip (LFS) assay for the rapid and specific detection of A. fuliginea. The RPA-CRISPR/Cas12b-LFS assay provides visual results within 35 min and does not rely on expensive equipment. High specificity was demonstrated against other Amanita species and non-Amanita species, with a sensitivity of 31 pg of genomic DNA per reaction. In conclusion, the RPA-CRISPR/Cas12b-LFS assay presents a rapid, specific, sensitive, and convenient tool for identifying A. fuliginea, particularly suitable for primary healthcare institutions or remote areas.
灰花纹鹅膏是一种常见的致命蘑菇，因其易于误食且早期症状隐匿，致死率很高。对该物种进行快速、特异性鉴定对于临床诊断和治疗至关重要。然而，现有的灰花纹鹅膏核酸快速检测方法仍受特异性问题困扰。在此，我们开发了一种重组酶聚合酶扩增（RPA）结合CRISPR/Cas12b和侧向流动试纸条（LFS）的检测方法，用于快速、特异性检测灰花纹鹅膏。RPA-CRISPR/Cas12b-LFS检测方法在35分钟内即可提供可视化结果，且不依赖昂贵设备。该方法对其他鹅膏物种和非鹅膏物种具有高度特异性，每个反应的基因组DNA灵敏度为31 pg。总之，RPA-CRISPR/Cas12b-LFS检测方法是一种快速、特异、灵敏且便捷的灰花纹鹅膏鉴定工具，特别适用于基层医疗机构或偏远地区。
