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RH-RPA:基于RNase HII与重组酶聚合酶放大功能的结合,用于核酸检测的快速和高度敏感的测试

Yuting Jin¹, Yangnan Fu¹, Qingyang Liu¹

¹State Key Laboratory of Cellular Stress Biology, Innovation Center for Cell Signaling Network, Engineering Research Centre of Molecular Diagnostics of the Ministry of Education, National Institute for Data Science in Health and Medicine Engineering, School of Life Sciences, Faculty of Medicine and Life Sciences, Shenzhen Research Institute of Xiamen University, Xiamen University, Xiamen, Fujian 361102, China.

Analytical chemistry

|April 24, 2025

概括

一种新的基于RNase HII的重组酶聚合酶放大 (RH-RPA) 试验提供了灵敏,快速和无设备的核酸检测. 这种分子诊断平台在临床应用中表现出高准确度.

科学领域:

分子生物学分子生物学
生物技术是生物技术.
诊断检测试验诊断检测试验

背景情况:

复合聚合酶放大 (RPA) 探针测定,包括RPA-exo和RPA-nfo,是目标检测的标准.
与RPA-exo相比,RPA-nfo的灵敏度较低,而RPA-exo缺乏无设备可视化,限制了更广泛的应用.
现有的方法在敏感度,可视化和核酸检测的广泛适用性方面存在局限性.

研究的目的:

开发一种与RNase HII (RH-RPA) 系统相结合的新型重组酶聚合酶放大 (RPA) 试验.
创建一个多功能分子诊断平台,支持光和侧流带检测.
克服现有的基于RPA的探针测定对增强核酸检测的局限性.

主要方法:

开发了一种新的RH-RPA测定方法,集成RNase HII用于目标识别和分离.
实现的双检测模式:光信号检测和侧流带读数.
使用DNA和RNA目标验证了测试的性能,包括用于检测流感A病毒的临床样本.

主要成果:

RH-RPA在20分钟内 (光模式) 实现了低至10个DNA/RNA拷贝/反应的检测极限.
侧流条形模式在20分钟内检测到只有5个拷贝/核酸反应.
RT RH-RPA在临床样本中检测流感A病毒的准确性达到100%.

结论:

RH-RPA提供了一个快速,高度敏感和特定的分子诊断平台.
该方法具有成本效益,适合于护理点核酸检测.
RH-RPA解决了以前的RPA分析的局限性,提高了诊断能力.

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