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在CRISPR/Cas9中介的T4菌基因组编辑中进行高通量抗菌感应性测试

Yawen He¹, Juhong Chen^1,2

¹Department of Biological Systems Engineering, Virginia Tech, Blacksburg, Virginia 24061, United States.

Analytical chemistry

|October 30, 2024

概括

一种基于菌体的新型色度检测策略可以准确地确定抗菌素耐药性. 该方法使用工程T4菌体进行快速抗微生物敏感性测试 (AST),为临床诊断提供视觉和高通量替代方案.

科学领域:

微生物学微生物学
生物技术是生物技术.
分子生物学分子生物学

背景情况:

准确的抗菌素耐药性测定对于控制传染病至关重要.
传统的抗微生物敏感性测试 (AST) 方法可能耗时.
在临床环境中需要快速,高吞吐率的AST方法.

研究的目的:

为AST开发一种简单,准确和高通量基色度感测策略.
使用CRISPR/Cas9系统为生物感知应用设计T4菌体.
提供一种可视和快速的方法来确定细菌的抗菌素敏感性.

主要方法:

T4菌体基因组的模块化工程,以携带lacZ-α (lacZa) 标记基因.
使用CRISPR/Cas9系统进行菌体基因组修改.
采用色度基质检测β-银酸酶活性,表明细菌生长和易感性.

主要成果:

成功开发和验证了一种基于菌体的色度测量策略,用于AST.
在尿样中检测出多抗药性大肠杆菌 (E. coli) 的100%特异性.
与传统的磁盘扩散测试相比,检测时间至少缩短了一半,而不会影响灵敏度.

结论:

基于菌体的工程传感策略为AST提供了一个快速,可视和高通量替代方案.
这种方法可以显著减少临床诊断中的检测时间.
该战略在识别多药耐药细菌方面表现出高度准确性和特异性.

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