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一种基于微流体卡的电化学试验,用于检测硫胺耐药基因

Patrick Severin Sfragano¹, Eduardo Canek Reynoso², Norma Elena Rojas-Ruíz³

¹Department of Chemistry "Ugo Schiff", University of Florence, 50019, Sesto Fiorentino, Italy.

|February 1, 2024

概括

本研究介绍了一种用于检测抗生素耐药性基因 (sul1和sul4) 的微流体卡上的快速,低成本的电化学测定方法. 该平台需要最小的专业知识和耗材,使抗菌素耐药性监测更快.

科学领域:

电化学电化学电化学
微流体学微流体学
分子生物学分子生物学
抗微生物耐药性抗微生物耐药性

背景情况:

传统的DNA标记物检测是耗时的,需要专家人员.
抗菌素耐药性 (AMR) 是一个全球性的健康威胁,需要快速的分析方法.
现有的检测AMR基因的方法往往复杂且资源密集.

研究的目的:

开发一种易于使用,低成本的电化学试验,用于检测硫胺抗性基因 (sul1和sul4).
整合微流体操纵和双放大策略,以提高检测灵敏度.
在临床和环境样本中提供一个快速有效的AMR监测平台.

主要方法:

开发基于卡片的微流体平台,用于自动化样本处理.
利用DNA修饰的磁粒子来捕获和分离放大产品.
在37°C使用酶辅助的异热放大用于DNA放大.
合样本/信号双放大,以改善检测.

主要成果:

达到的检测极限为硫1的44.2 pmol/L和硫4.5的48.5 pmol/L.
在大约1小时内成功检测到来自大肠杆菌细胞的硫胺耐药基因.
证明能够检测到≥500倍稀释的sul1.1的放大产品.
该平台需要最小的手动干预和小样本量.

结论:

开发的微流体电化学试验为检测AMR基因提供了快速,灵敏和经济有效的解决方案.
这个平台大大减少了DNA分析所需的实践时间和专业知识.
该系统有望实现分散的AMR监测和护理点诊断.

关键词:

抗微生物耐药性抗微生物耐药性基因传感器是一种基因传感器.异热放大器的同热放大方式磁性颗粒是一种磁性粒子.微流体学微流体学复合酶聚合酶放大复合酶的放大.

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