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超敏感的CRISPR/Cas12a基系统用于检测抗生素耐药微生物中的OXA-1基因

Marina Tyumentseva¹, Aleksandr Tyumentsev¹, Anna Prelovskaya¹

¹Central Research Institute of Epidemiology, Novogireevskaya Str., 3a, 111123 Moscow, Russia.

Current issues in molecular biology

|July 23, 2025

概括

一种新的CRISPR/Cas12a试验与PCR相结合,可以快速检测OXA-1基因,这对于识别抗生素耐药细菌至关重要. 这种快速检测方法有助于监测和打击危险抗菌素耐药性的传播.

科学领域:

微生物学微生物学
分子生物学分子生物学
生物技术是生物技术.

背景情况:

bla_OXA-1基因在扩展光谱β-乳糖酶 (ESBL) 生产微生物中赋予了对西林的耐药性.
这种基因在各种细菌家族中普遍存在,包括Enterobacteriaceae和Pseudomonadaceae.
目前对OXA-1的检测方法通常是缓慢且资源密集的.

研究的目的:

开发一种快速,灵敏的检测方法,用于检测携带bla_OXA-1基因的微生物.
利用CRISPR/Cas12a技术,加强对抗菌素耐药性的监测.

主要方法:

开发了一种结合聚合酶链反应 (PCR) 与CRISPR/Cas12a技术的新型检测方法.
该试验涉及使用PCR进行65分钟的预放大步骤,随后进行5分钟的基于CRISPR/Cas12a的检测步骤.
检测依赖于Cas12a酶的水解单链DNA光记者的能力.

主要成果:

该测试显示出高灵敏度,能够检测到bla_OXA-1基因的1.25个副本.
在模型系统和实际bla_OXA-1-阳性微生物中都实现了有效的检测.
整个检测过程比现有方法快得多.

结论:

开发的PCR合CRISPR/Cas12a试验提供了一种快速,灵敏和高效的方法来检测含有bla_OXA-1的微生物.
这种测试有可能改善对抗生素耐药细菌的监测.
加强监测可以有助于减轻抗菌素耐药性的全球威胁.

关键词:

这就是CRISPR/Cas12a.这就是ESBL.这是一个PCRPCR.抗生素耐药的微生物 blaOXA-1-1 的使用情况.检测检测检测检测检测

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