摘要

A PCR-free assay for rapid pathogen diagnosis was implemented on an integrated microfluidic system in this study. Vancomycin-conjugated magnetic beads were used to capture multiple strains of bacteria and nucleotide probes labeled gold nanoparticles were used to specify and detect a specific strain by hybridization-induced color change. The assay was entirely automated within an integrated microfluidic device that was composed of suction-type micropumps, microvalves, microchannels, and microchambers that fabricated by microfluidic technology. Multiple strains of bacteria could be captured simultaneously by vancomycin-conjugated magnetic beads, with capturing efficiency exceeding 80%. Subsequently, sensitive and strain-specific detection against target bacteria could be achieved by using nanogold labeled specific nucleotide probes. The limit of detection of 10(2) CFU bacteria was achieved. Importantly, nucleic acid amplification was not involved in the diagnostic procedures; the entire analytic process required only 25 min. The developed platform may provide a promising tool for rapid diagnosis of bacterial infections. From the Clinical Editor: In this novel study, a PCR-free pathogen detection method is demonstrated. After vancomycin-conjugated magnetic beads captured bacteria, nucleotide probes-labeled gold nanoparticles were employed to specify and detect specific strains via hybridization-induced color change. Multiple strains of bacteria could be captured simultaneously with an efficiency exceeding 80%, enabling the detection of as low as 10(2) CFU of bacteria.