食品科学 ›› 2023, Vol. 44 ›› Issue (24): 323-328.doi: 10.7506/spkx1002-6630-20230404-029

• 安全检测 • 上一篇    下一篇

基于裁剪适配体的纳米金比色法用于金黄色葡萄球菌肠毒素A的可视化检测

崔丽伟, 魏荣, 常惟丹, 岳晓禹, 许文涛   

  1. (1.河南牧业经济学院食品与生物工程学院,河南 郑州 450046;2.中国农业大学营养与健康系,食品精准营养与质量控制教育部重点实验室,北京 100191)
  • 出版日期:2023-12-25 发布日期:2024-01-02
  • 基金资助:
    “十四五”国家重点研发计划重点专项(2022YFF0607900);国家自然科学基金面上项目(32372437); 河南省科技攻关项目(212102310259;232102320281;232102320285);河南省高等学校重点科研项目(22A550012)

Tailored Aptamer-Based Visual Colorimetric Detection of Staphylococcal Enterotoxin A Using Gold Nanoparticles

CUI Liwei, WEI Rong, CHANG Weidan, YUE Xiaoyu, XU Wentao   

  1. (1. School of Food and Biological Engineering, Henan University of Animal Husbandry and Economy, Zhengzhou 450046, China; 2. Key Laboratory of Precision Nutrition and Food Quality, Ministry of Education, Department of Nutrition and Health, China Agricultural University, Beijing 100191, China)
  • Online:2023-12-25 Published:2024-01-02

摘要: 目的:筛选适配体作为分子识别原件,基于纳米金的盐效应构建生物传感器,实现金黄色葡萄球菌肠毒素A(staphylococcal enterotoxin A,SEA)的快速定量检测。方法:采用分子对接和分子动力学模拟对SEA适配体进行截短指导和结果预测,利用纳米金显色法验证模拟结果,对SEA适配体优化并将其作为分子识别原件,优化反应体系,探讨SEA质量浓度与吸光度比值间的关系,构建一种可视化的SEA快速检测分析方法,并对方法的准确度、精密度和特异性进行评价。结果:筛选出一条序列短、亲和力高、特异性强且稳定的SEA适配体,建立了一种基于纳米金比色的SEA可视化检测方法,检出限低,加标回收结果满意。结论:采用分子模拟能有效提高适配体筛选效率,本方法可用于SEA快速检测。

关键词: 金黄色葡萄球菌肠毒素A;可视化;适配体;裁剪;分子模拟;纳米金比色

Abstract: Objective: To select and use aptamers as molecular recognition elements to construct a biosensor based on the salt effect of gold nanoparticles for the rapid quantitative detection of staphylococcal enterotoxin A (SEA). Methods: Molecular docking and molecular dynamics simulation were used to truncate the SEA aptamer and predict the results. The simulation results were verified colorimetrically using gold nanoparticles. The optimal SEA aptamer was used as the molecular recognition element. The reaction system was optimized, the relationship between SEA mass concentration and absorbance ratio was explored, and the accuracy, precision and specificity of the developed method were evaluated. Results: An SEA aptamer with short sequence, high affinity, strong specificity and stability was selected. This method was characterized by low limit of detection (LOD) and satisfactory recoveries for spiked samples. Conclusion: Molecular simulation can effectively improve the screening efficiency of aptamers, and the proposed method can be used for rapid detection of SEA.

Key words: staphylococcal enterotoxin A; visualization; aptamer; truncation; molecular dynamics simulation; gold nanoparticles-based colorimetry

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