Self-Interference Digital Optofluidic Genotyping for Integrated and Automated Label-Free Pathogen Detection

• DOI: 10.1021/acssensors.4c01520
• Publication Date: 2024-11-19T20:38:49Z
• Authors: Tianqi Zhou; Rongxin Fu; Jialu Hou; Fan Yang; Fengli Chai; Zeyin Mao; Anni Deng; Fenggang Li; Yanfang Guan; Hanqi Hu; Hang Li; Yao Lu; Guoliang Huang; Shuailong Zhang; Huikai Xie
• Keywords: Recombinase Polymerase Amplification; Molecular diagnostics

Pathogen, prevalent in both natural and human environments, cause approximately 4.95 million deaths annually, ranking them among the top contributors to global mortality. Traditional pathogen detection methods, reliant on microscopy cultivation, are slow labor-intensive often produce subjective results. While nucleic acid amplification techniques such as polymerase chain reaction offer genetic accuracy, they necessitate costly laboratory equipment skilled personnel. Consequently, isothermal methods like recombinase (RPA) have attracted interest for their rapid straightforward operations. However, these face challenges specificity automated sample processing. In this study, we introduce a self-interferometric digital optofluidic platform incorporating asymmetric direct solid-phase RPA real-time, label-free, genotyping. By integration of microfluidics with DNA monolayer method using hyperspectral interferometry, enables rapid, specific, sensitive without need exogenous labeling or complex procedures. The system demonstrated high sensitivity (10 CFU·mL–1), (differentiating four Candida species), efficiency (fully within 50 min Gram-negative bacteria), throughput (simultaneous indices). This integrated approach quantitation single microfluidic chip represents significant advancement diagnostics, providing practical solution timely analysis.