Field-Deployable Detection of Genetically Modified Organisms with an Integrated Method of Loop-Mediated Isothermal Amplification and CRISPR/FnCas12a
文献类型： 外文期刊
作者： Chen, Lu 1 ; Wang, Chen 2 ; Zhu, Zaobing 3 ; Yang, Litao 1 ;
作者机构： 1.Shanghai Jiao Tong Univ, Yazhou Bay Inst Deepsea Sci Tech, Sanya 572024, Peoples R China
2.Shanghai Jiao Tong Univ, Sch Life Sci & Biotechnol, Joint Int Res Lab Metab & Dev Sci, Shanghai 200240, Peoples R China
3.Jiangsu Acad Agr Sci, Inst Plant Protect, Nanjing 210014, Peoples R China
关键词： Cas-pfLAMP; DBN9936; MON810; GTS 40-3-2; on-site detection; LAMP; CRISPR/Cas12a; lateral flow strip
期刊名称：JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY （ 影响因子：6.2； 五年影响因子：6.4 ）
ISSN： 0021-8561
年卷期： 2025 年 73 卷 9 期
页码：
收录情况： SCI
摘要： The detection of genetically modified organisms (GMOs) is crucial for regulatory compliance and consumer safety. This study presents a novel method combining loop-mediated isothermal amplification (LAMP) with CRISPR/Cas12a cleavage, termed Cas-pfLAMP, for sensitive and specific GMO detection. We developed assays for three GM events: maize DBN9936 and MON810 and soybean GTS40-3-2. By incorporating a universal protospacer adjacent motif (PAM) sequence into LAMP primers, we overcame the limitations of PAM site dependence. The Cas-pfLAMP assays demonstrated high specificity and sensitivity, with limits of detection as low as 10-12 copies per reaction. Furthermore, we developed a point-of-care testing platform integrating rapid DNA extraction, Cas-pfLAMP, and lateral flow strips for on-site GMO detection. This platform achieved comparable sensitivity to qPCR, detecting GM contents as low as 0.1% in simulated samples within 40 min. The Cas-pfLAMP method offers the advantages of PAM site independence, high specificity and sensitivity, and suitability for field testing without specialized equipment. This approach represents a promising new generation of GMO detection methods with potential applications in various scenarios.
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