A fluorescence ELISA based on dsDNA-CuNCs for the detection of E. coli O157:H7 in milk

Authors

CAO Wenkai, College of Food Science, Nanchang University, Nanchang, Jiangxi 330047, China
SHAN Shan, College of Life Science, Jiangxi Normal University, Nanchang, Jiangxi 330022, China
LIU Daofeng, Jiangxi Provincial Center for Disease Control and Prevention, Nanchang, Jiangxi 330029, China;Jiangxi Provincial Key Laboratory of Diagnosing and Tracing of Foodborne Disease, Nanchang, Jiangxi 330029, China
PENG Juan, College of Food Science, Nanchang University, Nanchang, Jiangxi 330047, China
XING Keyu, School of Food Science and Bioengineering, Changsha University of Science & Technology, Changsha, Hunan 410114, China
LAI Weihua, College of Food Science, Nanchang University, Nanchang, Jiangxi 330047, ChinaFollow

Corresponding Author(s)

赖卫华（1968—），男，南昌大学教授，博士。E-mail：talktolaiwh@163.com

Abstract

Objective: This study aimed to developed a novel fluorescent enzyme-linked immunosorbent assay method for the sensitive detection of E. coli O157:H7 in milk samples. Methods: The pyrophosphate-Cu²⁺ coordination complex was hydrolyzed by alkaline phosphatase, and Cu²⁺ was released from copper nanoclusters as signal reporting probes. Key factors such as Cu²⁺ concentration, pyrophosphate concentration, and DNA template concentration and length were optimized. Results: Under the control of the optimized conditions, the method exhibited a good linear relationship (R²=0.980 8) with the target E. coli O157:H7 in the range of 5×10⁴~1×10⁸ CFU/mL. The limit of detection was 2.4×10⁴ CFU/mL. Conclusion: The method was successfully applied to the determination of E. coli O157:H7 in samples of low-fat milk and skim milk. The average recovery was 92.2% ~ 108.5% with relative standard deviation 4.9% ~ 10.4%.

Publication Date

10-30-2023

First Page

38

Last Page

43,94

DOI

10.13652/j.spjx.1003.5788.2023.80734

Recommended Citation

Wenkai, CAO; Shan, SHAN; Daofeng, LIU; Juan, PENG; Keyu, XING; and Weihua, LAI (2023) "A fluorescence ELISA based on dsDNA-CuNCs for the detection of E. coli O157:H7 in milk," Food and Machinery: Vol. 39: Iss. 9, Article 6.
DOI: 10.13652/j.spjx.1003.5788.2023.80734
Available at: https://www.ifoodmm.cn/journal/vol39/iss9/6

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