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A Thermostable Cas12b-Powered Bioassay Coupled with Loop-Mediated Isothermal Amplification in a Customized "One-Pot" Vessel for Visual, Rapid, Sensitive, and On-Site Detection of Genetically Modified Crops

文献类型： 外文期刊

作者： Han, Xiao 1 ; Lu, Minghui 1 ; Zhang, Yaru 1 ; Liu, Xinru 1 ; Zhang, Qiang 1 ; Bai, Xue 3 ; Man, Shuli 1 ; Zhao, Liangjuan 4 ; Ma, Long 1 ;

作者机构： 1.Tianjin Univ Sci & Technol, Coll Biotechnol, State Key Lab Food Nutr & Safety, Key Lab Ind Microbiol,Minist Educ,Tianjin Key Lab, Tianjin 300457, Peoples R China

2.Branch Tianjin Third Cent Hosp, Tianjin 300457, Peoples R China

3.Chinese Acad Agr Sci, Inst Food Sci & Technol, Key Lab Agroprod Proc, Minist Agr, Beijing 100193, Peoples R China

4.Tianjin Normal Univ, Tianjin Key Lab Anim & Plant Resistance, Tianjin 300387, Peoples R China

关键词： BrCas12b; genetically modified crops (GMCs); CaMV35S promoter; one-pot; portable detection; food detection

期刊名称：JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY （ 影响因子：6.1； 五年影响因子：6.3 ）

ISSN： 0021-8561

年卷期： 2024 年 72 卷 19 期

页码：

收录情况： SCI

摘要： Genetically modified crops (GMCs) have been discussed due to unknown safety, and thus, it is imperative to develop an effective detection technology. CRISPR/Cas is deemed a burgeoning technology for nucleic acid detection. Herein, we developed a novel detection method for the first time, which combined thermostable Cas12b with loop-mediated isothermal amplification (LAMP), to detect genetically modified (GM) soybeans in a customized one-pot vessel. In our method, LAMP-specific primers were used to amplify the cauliflower mosaic virus 35S promoter (CaMV35S) of the GM soybean samples. The corresponding amplicons activated the trans-cleavage activity of Cas12b, which resulted in the change of fluorescence intensity. The proposed bioassay was capable of detecting synthetic plasmid DNA samples down to 10 copies/mu L, and as few as 0.05% transgenic contents could be detected in less than 40 min. This work presented an original detection method for GMCs, which performed rapid, on-site, and deployable detection.

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