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

Dong-Ku Kang, M. Monsur Ali, Kaixiang Zhang, Susan S. Huang, Ellena Peterson, Michelle A. Digman, Enrico Gratton and Weian Zhao ()
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Dong-Ku Kang: University of California–Irvine
M. Monsur Ali: University of California–Irvine
Kaixiang Zhang: University of California–Irvine
Susan S. Huang: School of Medicine, University of California–Irvine
Ellena Peterson: University of California
Michelle A. Digman: University of California–Irvine
Enrico Gratton: University of California–Irvine
Weian Zhao: University of California–Irvine

Nature Communications, 2014, vol. 5, issue 1, 1-10

Abstract: Abstract 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.

Date: 2014
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DOI: 10.1038/ncomms6427

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