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

• Published in: Analytical chemistry (Washington) Vol. 96; no. 16; pp. 6282 - 6291
• Main Authors: Xiang, Xinran, Ren, Xiaoqing, Wen, Qianyu, Xing, Gaowa, Liu, Yuting, Xu, Xiaowei, Wei, Yuhuan, Ji, Yuhan, Liu, Tingting, Song, Huwei, Zhang, Shenghang, Shang, Yuting, Song, Minghui
• Format: Journal Article
• Language: English
• Published: United States American Chemical Society 23.04.2024
• Subjects: Automation; Bacteria; Bacteria - genetics; Bacteria - isolation & purification; Bacterial Infections - diagnosis; Clustered Regularly Interspaced Short Palindromic Repeats - genetics; CRISPR; Diagnostic systems; Humans; Lab-On-A-Chip Devices; Microfluidic Analytical Techniques - instrumentation; Microfluidics; Nucleic Acid Amplification Techniques; Pathogens; Recombinase; Recombinases - metabolism; Respiratory diseases; Respiratory disorders; Respiratory tract; Respiratory tract infection; Respiratory Tract Infections - diagnosis; Respiratory Tract Infections - microbiology; System effectiveness
• ISSN: 0003-2700; 1520-6882; 1520-6882
• DOI: 10.1021/acs.analchem.3c05682

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.