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超敏感的双重ELONA/SERS-RPA多重诊断对抗生素耐药性的诊断

Waleed A Hassanain¹, Christopher L Johnson², Karen Faulds¹

¹Department of Pure and Applied Chemistry, Technology and Innovation Centre, University of Strathclyde, Glasgow G1 1RD, U.K.

Analytical chemistry

|July 8, 2024

概括

本研究引入了一种基于磁珠的快速测定方法,使用复合酶聚合酶放大 (RPA) 来检测抗菌素耐药性 (AMR) 基因. 这种新的方法在两个小时内提供了灵敏的临床检测,有助于对抗抗性感染.

科学领域:

分子生物学分子生物学
生物技术是生物技术.
医学诊断医学诊断医学诊断

背景情况:

抗微生物药物耐药性 (AMR) 构成一个关键的全球健康挑战,需要精确的抗微生物药物处方和有效的诊断工具.
目前的诊断方法往往缺乏及时临床决策和AMR监测所需的速度和灵敏度.
快速识别耐药病原体对于改善患者的治疗结果和制AMR的传播至关重要.

研究的目的:

开发和验证一种新的,快速和敏感的诊断平台,用于检测与AMR相关的关键碳酶基因 (VIM,KPC,IMP).
整合基于磁珠的丰富,重组酶聚合酶放大 (RPA) 和双检测方法 (ELONA和SERS) 进行增强的DNA分析.
为了展示该平台在AMR标志物的多重检测和护理点检测方面的潜力.

主要方法:

用生物化原料为目标基因放大而涂上斯特雷普塔维丁的磁珠的功能化.
在磁支架上的异热再组合酶聚合酶放大 (RPA) 产生固定,基因特异的amplicons.
通过使用HRP探针和使用选择性探针的表面增强拉曼散射 (SERS) 检测用酶链 oligonucleotide assay (ELONA) 检测安普利康.
在单个反应管中多重检测VIM,KPC和IMP基因.

主要成果:

在ELONA和SERS检测试验中实现了亚皮科莫拉量化极限,使得早期诊断成为可能.
在单个管中成功检测了三种碳酶基因 (VIM,KPC,IMP) 的多重检测.
整个测定过程,从样品到结果,在不到2小时内完成.
该平台表现出高灵敏度,便携性和速度,适合于护理点应用.

结论:

开发的磁性支架ELONA/SERS-RPA平台为快速DNA检测和AMR监测提供了一个强大的工具.
该试验的速度,同热性和灵敏度使其成为传统PCR对临床诊断的有希望的替代品.
这项技术有潜力显著帮助医疗保健从业人员在护理点对抗抗菌素耐药性.

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