Automatic Microfluidic Harmonized RAA-CRISPR Diagnostic System for Rapid and Accurate Identification of Bacterial Respiratory Tract Infections

清脆的 计算生物学 微流控 微流控芯片 重组酶聚合酶扩增 纳米技术 聚合酶链反应 生物化学 基因 材料科学 生物
作者
Yi Hu,Xiaoqing Ren,Qianyu Wen,Gaowa Xing,Yuting Liu,Xiaowei Xu,Yuxi Wei,Yuhan Ji,Tingting Liu,Huwei Song,Shenghang Zhang,Yuting Shang,Minghui Song
出处
期刊:Analytical Chemistry [American Chemical Society]
卷期号:96 (16): 6282-6291
标识
DOI:10.1021/acs.analchem.3c05682
摘要

Respiratory tract infections (RTIs) pose a grave threat to human health, with bacterial pathogens being the primary culprits behind severe illness and mortality. In response to the pressing issue, we developed a centrifugal microfluidic chip integrated with a recombinase-aided amplification (RAA)-clustered regularly interspaced short palindromic repeats (CRISPR) system to achieve rapid detection of respiratory pathogens. The limitations of conventional two-step CRISPR-mediated systems were effectively addressed by employing the all-in-one RAA-CRISPR detection method, thereby enhancing the accuracy and sensitivity of bacterial detection. Moreover, the integration of a centrifugal microfluidic chip led to reduced sample consumption and significantly improved the detection throughput, enabling the simultaneous detection of multiple respiratory pathogens. Furthermore, the incorporation of Chelex-100 in the sample pretreatment enabled a sample-to-answer capability. This pivotal addition facilitated the deployment of the system in real clinical sample testing, enabling the accurate detection of 12 common respiratory bacteria within a set of 60 clinical samples. The system offers rapid and reliable results that are crucial for clinical diagnosis, enabling healthcare professionals to administer timely and accurate treatment interventions to patients.
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