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高负载辣根过氧化物酶和生物识别抗体超分子自组装成金/聚多巴胺纳米复合材料用于牛奶中大肠杆菌O157:H7的灵敏免疫测定

Supramolecular self-assembly of high-loaded horseradish peroxidase and biorecognized antibody into Au/polydopamine nanocomposites for sensitive immunoassay of Escherichia coli O157:H7 in milk.

作者信息

Fang Bolong, Li Yuzhi, Zhao Xiaole, Liu Yan, Xu Lin, Wang Qiao, Liu Xin, Gong Zhiyong, Lai Weihua

机构信息

Hubei Key Laboratory for Processing and Transformation of Agricultural Products, College of Food Science and Engineering, Wuhan Polytechnic University, Wuhan 430023, China.

Hubei Key Laboratory for Processing and Transformation of Agricultural Products, College of Food Science and Engineering, Wuhan Polytechnic University, Wuhan 430023, China.

出版信息

J Dairy Sci. 2025 Apr;108(4):3419-3427. doi: 10.3168/jds.2024-25518. Epub 2025 Jan 29.

Abstract

There is an urgent need for a rapid and sensitive method for the detection of Escherichia coli O157:H7, a class of hazardous foodborne pathogens in food safety. The traditional ELISA, a dominant rapid detection technique, has the disadvantage of low test sensitivity due to the insufficient enzyme loading capacity. In this study, we successfully synthesized self-assembled Au/polydopamine (PDA)/horseradish peroxide (HRP) nanocomposites with high enzyme loading on the outer surface and in the inner space. The high catalytic activity of Au/PDA/HRP was maintained by virtue of its hyperbranched flexible structure. For E. coli O157:H7 detection in milk samples, the proposed immunoassay achieved a visual cut-off value of 10 cfu mL and a low limit of detection of 2.8 × 10 cfu mL, which are 33 and 46 times more sensitive than the traditional ELISA, respectively. The tremendous advantages of high sensitivity, excellent specificity, and adequate recovery make it promising for monitoring various kinds of pathogenic bacteria in food safety with greater sensitivity.

摘要

迫切需要一种快速灵敏的方法来检测大肠杆菌O157:H7,这是食品安全中一类有害的食源性病原体。传统的酶联免疫吸附测定法(ELISA)作为一种占主导地位的快速检测技术,由于酶负载能力不足而存在检测灵敏度低的缺点。在本研究中,我们成功合成了自组装的金/聚多巴胺(PDA)/辣根过氧化物酶(HRP)纳米复合材料,其外表面和内部空间均具有高酶负载量。Au/PDA/HRP因其超支化柔性结构而保持了高催化活性。对于牛奶样品中的大肠杆菌O157:H7检测,所提出的免疫测定法的目视截断值为10 cfu/mL,检测下限低至2.8×10 cfu/mL,分别比传统ELISA灵敏33倍和46倍。高灵敏度、优异特异性和足够回收率的巨大优势使其有望以更高的灵敏度监测食品安全中的各种病原菌。

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