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An Argonaute-mediated bio-barcode bioassay for one-tube and on-site detection of Staphylococcus aureus

2024; Elsevier BV; Volume: 410; Linguagem: Inglês

10.1016/j.snb.2024.135713

1873-3077

Yaru Li, Yajie Liu, Qiang Zhang, Xiaoqin Tang, Shuli Man, Shengying Ye, Long Ma,

Innovative Microfluidic and Catalytic Techniques Innovation

Isothermal amplification method has enabled major advancement toward rapid and deployable detection for nucleic acid. While exciting, there are still a series of deficiencies facing their practical implementation, such as carryover cross-contamination, which are major bottlenecks that must be addressed. In addition, there is a lack of high-throughput detection for foodborne pathogens in foods. Here, we reported an Argonaute-mediated bio-barcode bioassay for one-step cleavage, on-site and high-throughput detection of S. aureus (Staphylococcus aureus) through introducing Tag sequence and recombinase aided amplification (RAA). Furthermore, we designed a single vessel via tube-in-tube manner for one-tube contamination-free and portable detection without opening the lid. With this strategy, the same detection limit as fluorescence detection could be obtained, 1 CFU/mL with high specificity for S. aureus. We further evaluated the performance of bio-barcode biosensing strategy using S. aureus-contaminated clinical and food samples. Argonaute-mediated bio-barcode bioassay had the advantages of modular, universal plug-and-play and high-throughput scanning. This work not only satisfied the requirement for convenient detection but also fulfilled the need for portable on-site nucleic acid detection with portability, adaptability, transferability and practicality.