研究目的
To introduce a fluorescent, microfluidic assay that can characterize β-lactamase derived antibiotic susceptibility in 20 min with a sensitivity suitable for direct human specimens, providing point of care information for treatment guidance.
研究成果
The microfluidic β-LEAF assay provides a rapid, cost-effective, and simple diagnostic tool for characterizing β-lactamase derived antibiotic susceptibility, offering immediate treatment guidance at the point of care.
研究不足
The sensitivity of the assay may be limited by the background drift of the measurement, particularly with different β-lactamase enzymes having varying catalytic rates against the β-LEAF probe.
1：Experimental Design and Method Selection:
The study utilized a fluorescent, microfluidic assay (β-lactamase enzyme activated fluorophore or β-LEAF) designed to phenotypically detect β-lactamase derived antibiotic susceptibility.
2：Sample Selection and Data Sources:
Bacterial strains known to express β-lactamase and not express β-lactamase were used.
3：List of Experimental Equipment and Materials:
Included polymethyl methacrylate, nuclear track etched polycarbonate filters, and a vacuum aspirator for fluid flow.
4：Experimental Procedures and Operational Workflow:
The assay involved trapping bacteria directly from unprocessed patient samples using disposable microfluidic cartridges and monitoring fluorescent output.
5：Data Analysis Methods:
Fluorescence was measured to determine the rate of β-LEAF cleavage, indicating antibiotic susceptibility.
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