Portal de Periódicos da CAPES

Rapid detection of single bacteria in unprocessed blood using Integrated Comprehensive Droplet Digital Detection

2014; Nature Portfolio; Volume: 5; Issue: 1 Linguagem: Inglês

10.1038/ncomms6427

2041-1723

Dong‐Ku Kang, M. Monsur Ali, Kaixiang Zhang, Susan S. Huang, Ellena M. Peterson, Michelle A. Digman, Enrico Gratton, Weian Zhao,

Microfluidic and Capillary Electrophoresis Applications

Blood stream infection or sepsis is a major health problem worldwide, with extremely high mortality, which is partly due to the inability to rapidly detect and identify bacteria in the early stages of infection. Here we present a new technology termed 'Integrated Comprehensive Droplet Digital Detection' (IC 3D) that can selectively detect bacteria directly from milliliters of diluted blood at single-cell sensitivity in a one-step, culture- and amplification-free process within 1.5-4 h. The IC 3D integrates real-time, DNAzyme-based sensors, droplet microencapsulation and a high-throughput 3D particle counter system. Using Escherichia coli as a target, we demonstrate that the IC 3D can provide absolute quantification of both stock and clinical isolates of E. coli in spiked blood within a broad range of extremely low concentration from 1 to 10,000 bacteria per ml with exceptional robustness and limit of detection in the single digit regime.