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RPA-CRISPR/Cas12a Combined with Rolling Circle Amplification- Enriched DNAzyme: A Homogeneous Photothermal Sensing Strategy for Plant Pathogens

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文献类型：外文期刊

作者： Liu, Yanlin ¹ ; Ma, Lanrui ¹ ; Liu, Wenjing ³ ; Xie, Longyingzi ¹ ; Wu, Qi ¹ ; Wang, Yiwen ¹ ; Zhou, Yan ¹ ; Zhang, Yaohai ¹ ; Jiao, Bining ¹ ; He, Yue ¹ ;

作者机构： 1.Southwest Univ, Key Lab Qual & Safety Control Citrus Fruits, Minist Agr & Rural Affairs, Chongqing 400712, Peoples R China

2.Southwest Univ, Citrus Res Inst, Natl Citrus Engn Res Ctr, Chongqing 400712, Peoples R China

3.Fujian Acad Agr Sci, Inst Agr Qual Stand & Testing Technol Res, Fujian Key Lab Agroprod Qualitiy & Safety, Fuzhou 350003, Peoples R China

关键词： recombinase polymerase amplification; CRISPR; Cas12a; rolling circle amplification; G-quadruplex; portable detection; plant pathogens

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： 2023 年 71 卷 11 期

页码：

收录情况： SCI

摘要： Alternaria is an endemic fungus associated with brown spot disease, which is one of the most serious citrus diseases. In addition, the mycotoxins metabolized by Alternaria threaten human health seriously. Herein, a novel homogeneous and portable qualitative photothermal method based on recombinase polymerase amplification (RPA), CRISPR/Cas12a, and rolling circle amplification (RCA) for the detection of Alternaria is described. Using RCA primers as substrates for CRISPR/Cas12a trans- cleavage, the two systems, RPA-CRISPR/Cas12a and RCA-enriched G-quadruplex/hemin DNAzyme, are intelligently combined. Target DNA at fg/mu L levels can be detected with high specificity. Additionally, the practicability of the proposed method is demonstrated by analyzing cultured Alternaria from different fruit and vegetable samples, as well as citrus fruit samples collected in the field. Furthermore, the implementation of this method does not require any sophisticated equipment and complicated washing steps. Therefore, it has great potential to screen Alternaria in poor laboratories.

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RPA-CRISPR/Cas12a Combined with Rolling Circle Amplification- Enriched DNAzyme: A Homogeneous Photothermal Sensing Strategy for Plant Pathogens

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