Rapid detection of single bacteria in unprocessed blood using Integrated Comprehensive Droplet Digital Detection.
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.
1] 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.
Dong-Ku
DK
Kang
1] 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.
M Monsur
MM
Ali
1] 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 [6] Department of Chemistry, Beijing Key Laboratory for Analytical Methods and Instrumentation, Tsinghua University, Beijing 100084, China.
Kaixiang
K
Zhang
Division of Infectious Diseases and Health Policy Research Institute, School of Medicine, University of California-Irvine, Irvine, California 92697, USA.
Susan S
SS
Huang
Department of Pathology and Laboratory Medicine, University of California, Irvine, California 92697, USA.
Ellena
E
Peterson
1] Department of Biomedical Engineering, University of California-Irvine, 845 Health Sciences Road, Irvine, California 92697, USA [2] Laboratory for Fluorescence Dynamics, University of California, Irvine, California 92697, USA [3] Centre for Bioactive Discovery in Health and Ageing, School of Science and Technology, University of New England, Armidale, New South Wales 2351, Australia.
Michelle A
MA
Digman
1] Department of Biomedical Engineering, University of California-Irvine, 845 Health Sciences Road, Irvine, California 92697, USA [2] Laboratory for Fluorescence Dynamics, University of California, Irvine, California 92697, USA.
Enrico
E
Gratton
1] 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.
Weian
W
Zhao
D001769:Blood / Q000382:microbiology*; D004269:DNA, Bacterial / Q000097:blood; D021881:DNA, Catalytic / Q000378:metabolism; D004339:Drug Compounding / Q000379:methods; D004926:Escherichia coli / Q000235:genetics; D006801:Humans; D057230:Limit of Detection; D046210:Microfluidic Analytical Techniques / Q000379:methods; D012680:Sensitivity and Specificity; D018805:Sepsis / Q000097:blood / Q000175:diagnosis* / Q000382:microbiology
D004269:DNA, Bacterial; D021881:DNA, Catalytic