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Ultrasensitive and visual identification of lethal mushroom Russula subnigricans by LAMP-based CRISPR/Cas12a assay

文献类型： 外文期刊

作者： Zhao, Zhiyong 1 ; Zhao, Lanxin 1 ; Tian, Enjing 2 ; Zheng, Fenshuang 3 ; Hengchao, E. 1 ; Zhao, Xiaoyan 1 ; Zhou, Changyan 1 ;

作者机构： 1.Shanghai Acad Agr Sci, Inst Agrofood Stand & Testing Technol, Lab Qual & Safety Risk Assessment Agroprod Shangha, Minist Agr & Rural Affairs, 1000 Jinqi Rd, Shanghai 201403, Peoples R China

2.Jilin Agr Univ, Engn Res Ctr, Chinese Minist Educ Edible & Med Fungi, Jilin 130118, Peoples R China

3.Yunnan Univ, Yunnan Peoples Hosp 2, Nucl Med, Affiliated Hosp, 176 Qingnian Rd, Kunming 650021, Peoples R China

4.Shanghai Guosen Biotechnol Co Ltd, Shanghai 201400, Peoples R China

关键词： Rapid detection; LAMP; CRISPR/Cas12a; Russula subnigricans; High sensitivity

期刊名称：MICROCHEMICAL JOURNAL （ 影响因子：5.1； 五年影响因子：4.7 ）

ISSN： 0026-265X

年卷期： 2025 年 211 卷

页码：

收录情况： SCI

摘要： Russula subnigricans is a lethal mushroom that causes rhabdomyolysis. Misidentification of the mushroom species can lead to serious foodborne poisoning with a mortality rate exceeding 50 %, posing a serious threat to human health. A sensitive and rapid detection strategy is urgently needed. In this study, we developed a loop-mediated isothermal amplification (LAMP) coupled with CRISPR/Cas12a assay for the accurate identification of lethal R. subnigricans. The assay could be completed in 70 min without the need for specialized equipment, and the results could be directly observed using the naked eye under UV light. The method exhibited high specificity with no cross-reaction in 22 other mushroom species. Sensitivity evaluation showed that the detection limits were as low as 1 fg/mu L of target DNA or 0.01 % mushroom admixture, presenting 500-1000 times higher sensitivity compared to the previously reported methods. The developed assay provides a valuable tool for rapid identification of R. subnigricans in remote, resource-limited rural areas to prevent the mushroom poisoning occurrence.

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