Digital genotyping of single pathogenic bacteria directly from nasal swab samples


20.-22. February | Ilmenau, Elgersburg
Talk by Felix von Stetten at the 9th Workshop of Chemical and Biological Micro Laboratory Technology in Ilmenau, Elgersburg (Thuringia, Germany)


The centrifugal microfluidic platform has demonstrated huge potential in automation of molecular diagnostic assays at the point-of-need, ranging from infectious disease and cancer diagnostics, over veterinary diagnostics to food analysis. A novel unit operation, designated centrifugal step emulsification, augments the platform with the ability to perform digital assays. Compared to the state-of-the-art the novel approach excels through ultimate process integration, its compatibility to standard laboratory devices, and its amenability to sample-to-result testing at the point-of-need. As proof of concept, we demonstrate the digital analysis of antibiotic resistant pathogenic bacteria directly from nasal swabs. Efficiencies of the various steps are discussed, comprising (1) the elution of the bacteria from the nasal swab, (2) the encapsulation of single bacterial cells into droplets, (3) the droplet integrated cell lysis, and (4) the isothermal multiplex amplification with simultaneous detection of species and resistance genes. The single cell approach is suggested to improve target oriented antibiotic therapy. Our future research will extend to the further digital multistep and multianalyte assays with applications ranging from metabiome screening to allergen testing.

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