数字农科院2.0

A one-pot and one-step RPA-CRISPR/Cas12b platform applied for streamlined detection of Salmonella spp.

文献类型： 外文期刊

作者： Jiansen Gong;Linlin Zhuang;Lixia Fu;Huifang Yin;Xinhong Dou;Cuiqin Huang;Xiangan Han

作者机构：

关键词： Cas12b;invA;One-pot;One-step;RPA;Salmonella

期刊名称： Journal of Agriculture and Food Research

ISSN： 2666-1543

年卷期： 2025 年 23 卷

页码：

收录情况： ESCI（2025版）

摘要： Salmonella spp. remains a primary culprit behind foodborne illnesses, posing significant risks to human health and the economy. Existing detection methods for Salmonella are often hindered by several drawbacks. Here, a streamlined and precise detection approach was introduced for Salmonella, leveraging the integration of RPA technology with the CRISPR/Cas12b system. This combined RPA-CRISPR/Cas12b method operates in a single PCR tube, simplifying the workflow to a one-step process. Our findings indicate that this assay has a limit of detection (LOD) of 60 copies per test and can be completed within just 1 h. Importantly, the assay demonstrated 100 % specificity for Salmonella, without any cross-reactivity with other bacterial strains. In practical tests, positive amplification was achieved even in the case that Salmonella contamination level reached 19.5 CFU/ml. Furthermore, an equal number of positive poultry samples were detected by using traditional culture method and the present RPA-CRISPR/Cas12b method, including chicken (52/483, 28.42 %), duck (19/40, 47.50 %), goose (14/31, 45.16 %), and pigeon (10/52, 19.23 %), respectively. Overall, this research introduces a novel detection system for Salmonella and underscores the promising potential of this method in curbing foodborne diseases.

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