Wang Zhipan, Cheng Xinrui, Ma Aimin, Jiang Feng, Chen Yiping
College of Food Science and Technology, Huazhong Agricultural University, Wuhan, China.
Key Laboratory of Detection Technology of Focus Chemical Hazards in Animal-Derived Food for State Market Regulation, Wuhan, China.
Nat Food. 2025 Feb;6(2):170-181. doi: 10.1038/s43016-024-01082-y. Epub 2025 Jan 2.
Accurate, sensitive and multiplexed detection of food-borne pathogens is crucial for assessing food safety risks. Here we present a digital DNA-amplification-free nucleic acid detection assay to achieve multiplexed and ultrasensitive detection of three food-borne pathogens. We used mesophilic Clostridium butyricum argonaute and magnetic beads in a digital carrier system (d-MAGIC). Clostridium butyricum argonaute, with its two-guide accurate cleavage activity, precisely targets and cleaves fluorescence-quencher reporters corresponding to different bacteria through a two-step process. The system uses fluorescence-encoded magnetic beads as programmable multi-probes, allowing the simultaneous detection of multiple pathogens and easy data interpretation via artificial intelligence. The method showed a wide detection range (10 to 10 CFU ml) and a low limit of detection of 6 CFU ml for food-borne pathogens without DNA amplification. Digital nucleic acid testing using d-MAGIC can become a next-generation strategy for accurate and convenient pathogen detection.
准确、灵敏且能进行多重检测的食源性病原体检测对于评估食品安全风险至关重要。在此，我们展示了一种无数字DNA扩增的核酸检测方法，以实现对三种食源性病原体的多重超灵敏检测。我们在数字载体系统（d-MAGIC）中使用了嗜温丁酸梭菌Argonaute和磁珠。丁酸梭菌Argonaute具有双向导精确切割活性，通过两步过程精确靶向并切割对应不同细菌的荧光淬灭报告分子。该系统使用荧光编码磁珠作为可编程多探针，能够同时检测多种病原体，并通过人工智能实现数据的轻松解读。该方法显示出较宽的检测范围（10至10 CFU/ml），对于未进行DNA扩增的食源性病原体，检测下限低至6 CFU/ml。使用d-MAGIC的数字核酸检测可成为准确便捷的病原体检测的下一代策略。
