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Rapid detection of single bacteria in unprocessed blood using Integrated Comprehensive Droplet Digital Detection
Dong-Ku Kang1, M Monsur Ali1, Kaixiang Zhang2
11] Department of Pharmaceutical Sciences, University of California-Irvine, 845 Health Sciences Road, Irvine, California 92697, USA [2] Sue and Bill Gross Stem Cell Research Center, University of California-Irvine, 845 Health Sciences Road, Irvine, California 92697, USA [3] Chao Family Comprehensive Cancer Center, University of California-Irvine, 845 Health Sciences Road, Irvine, California 92697, USA [4] Edwards Lifesciences Center for Advanced Cardiovascular Technology, University of California-Irvine, 845 Health Sciences Road, Irvine, California 92697, USA [5] Department of Biomedical Engineering, University of California-Irvine, 845 Health Sciences Road, Irvine, California 92697, USA.
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.
