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同热放大和CRISPR/Cas12a基于系统的测试用于快速,灵敏和视觉检测金黄色葡萄球菌

Danhong Xu¹, Haijuan Zeng^2,3, Wenhui Wu¹

¹School of Food Science and Technology, Shanghai Ocean University, Shanghai 201499, China.

Foods (Basel, Switzerland)

|December 23, 2023

概括

使用异热放大和CRISPR/Cas12a技术的新平台能够快速,灵敏地检测金黄色葡萄球菌. 这种方法在食品安全方面是有效的,在约40分钟内提供视觉或基于光的结果.

科学领域:

食品微生物学食品微生物学
分子诊断学分子诊断学
生物技术是生物技术.

背景情况:

金黄色葡萄球菌 (Staphylococcus aureus) 是一种常见的食源性病原体,会引起细菌病.
目前的S. aureus检测方法缺乏有效的食品安全管理所需的速度,特异性和敏感性.

研究的目的:

开发一种快速,高特异性和灵敏的金黄色葡萄球菌 (nuc基因) 检测平台.
将同热放大 (LAMP,RPA) 与CRISPR/Cas12a技术相结合,以有效检测细菌.

主要方法:

与CRISPR/Cas12a系统结合的循环介导同热放大 (LAMP) 或再组合聚合酶放大 (RPA) 的开发.
集成CRISPR/Cas12a平台与免疫染色学测试条 (ICS) 进行视觉检测.
对光检测平台 (LAMP,RPA-CRISPR/Cas12a-flu) 进行调整,以提高灵敏度.

主要成果:

核DNA的视觉检测极限为57.8 fg/μL,黄金菌的视觉检测极限为6.7 × 10^2 CFU/mL.
基因组DNA的光检测极限为5.78 fg/μL,黄金菌的CFU/mL为67 CFU/mL.
在大约40分钟内成功检测出牛奶产品中的金黄色细菌.

结论:

异热放大CRISPR/Cas12a平台提供了一个快速而灵敏的工具,用于检测食品中的金黄色细菌.
这项技术提供了一种低成本,简单,可视化的方法,适合食品安全应用.
该平台显示了广泛用于监测和控制食品中黄金菌污染的潜力.

关键词:

一个灯的灯光.在 RPA RPA 中使用.黄金色的S. 黄金色的光检测的检测方法免疫染色学试验带测试带.

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