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通过非对称的聚合酶链反应-Cas12a平台,从尿样直接进行快速细菌识别和抗菌敏感性测试

Laibao Zheng¹, Xinxi Zhou¹, Yan Zhang¹

¹Wenzhou Key Laboratory of Sanitary Microbiology, Key Laboratory of Laboratory Medicine, Ministry of Education, School of Laboratory Medicine and Life Sciences, Wenzhou Medical University, Wenzhou, Zhejiang 325035, China.

Analytical chemistry

|November 19, 2025

概括

一个新的诊断平台结合了不对称的PCR和CRISPR/Cas12a,用于快速细菌识别和直接从尿液中测试抗微生物敏感性. 这大大缩短了诊断时间,有助于感染控制和抗菌药物管理.

科学领域:

临床微生物学临床微生物学
分子诊断学分子诊断学
抗微生物耐药性抗微生物耐药性

背景情况:

抗菌素耐药性 (AMR) 是一个全球性的健康危机,由缓慢的常规诊断加剧.
目前用于病原体识别 (ID) 和抗微生物敏感性测试 (AST) 的方法超过48小时.
快速诊断对于有效的治疗和抗菌药物管理至关重要.

研究的目的:

开发和验证从尿样中直接检测细菌ID和表型AST的快速诊断平台.
显著减少诊断尿路感染的周转时间.
为临床微生物学提供具有成本效益和可扩展的解决方案.

主要方法:

不对称的聚合酶链反应 (aPCR) 与CRISPR/Cas12a技术的整合.
单复式aPCR针对16SrDNAV3-V4区域进行DNA放大.
CRISPR/Cas12a系统利用PCR产生的ssDNA进行多重病原体检测和表型AST.

主要成果:

在aPCR-Cas12a平台检测到六个常见的尿病原体在10^3CFU/mL.
现型AST在60分钟内完成,准确区分了敏感和耐药菌株.
通过86个临床尿样进行验证,与ID的培养完全一致,AST的高精度.
总工作流程时间从标准方法的>48小时减少到5.5小时.

结论:

aPCR-CRISPR/Cas12a平台为诊断尿路感染提供了一个快速,可靠和高效的方法.
这项技术显著缩短了诊断周转时间,支持及时的临床决策.
该平台显示了将其整合到常规临床微生物学和照顾点设置中的强大潜力,以改善抗菌药物管理.

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