Multiplex Aptamer-Based Fluorescence Assay Using Magnetism-Encoded Nanoparticles for Simultaneous Detection of Multiple Pathogenic Bacteria

Abstract

Multiplex assay has emerged as a robust and versatile method for the simultaneous detection of multiple analytes in a single test. However, challenges in terms of poor accuracy and complexity remained. In this work, we developed a multiplex aptamer-based fluorescence assay using magnetism-encoded nanoparticles for the simultaneous detection of multiple pathogenic bacteria. The encapsulation of different amounts of Fe 3 O 4 nanoparticles in zeolitic imidazolate framework-90 (ZIF-90) leads to the formation of Fe 3 O 4 @ZIF-90 (FZ) composites with distinct magnetism strengths. By functionalizing a specific aptamer on the surface of the FZ composites, target bacteria can be specifically and precisely separated from a mixed sample in a sequential manner. This property allows for the simultaneous quantitative analysis of multiple target bacteria by using a single-color fluorescence label, thereby resulting in minimal spectral crosstalk interference and improved accuracy. The successful determination of multiple bacteria in contaminated milk samples demonstrates the applicability of this multiplex assay in complex biological matrices. Compared to conventional multiplex fluorescence assays, this approach offers distinct advantages of simplicity, efficiency, and implementation. We believe that this study can provide valuable insights into the development of the multiplex assay while introducing a new method for the simultaneous detection of multiple bacteria.

Photoelectrochemical/Colorimetric Dual-Mode Specific Detection of Staphylococcus aureus Based on the Enzymatic Reaction Triggered by Catalase from Lysed Bacteria

“…Until now, most of the reported bacterial sensors were based on the biorecognition elements, such as aptamers, antibodies, vancomycin, bacteriophages, etc. Among them, bacteriophages are a class of viruses that specifically target bacteria without posing any harm to the human body, offer distinct advantages due to their abundance in nature, robust proliferative capabilities, cost-effective, and greater tolerance to pH and temperature variations, resulting in superior stability .…”