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Open AccessArticle

Rapid Detection of β-Lactamase-Producing Bacteria Using the Integrated Comprehensive Droplet Digital Detection (IC 3D) System

by Yiyan Li 1,†, Hemanth Cherukury 2,3,†, Louai Labanieh 4, Weian Zhao 2,3,5,6,7,8 and Dong-Ku Kang 9,10,*
1
Department of Physics and Engineering, Fort Lewis College, Durango, CO 81301, USA
2
Department of Pharmaceutical Sciences, University of California Irvine, Irvine, CA 92697, USA
3
Sue and Bill Gross Stem Cell Research Center, University of California Irvine, Irvine, CA 92697, USA
4
Department of Bioengineering, Stanford University, Stanford, CA 94305, USA
5
Chao Family Comprehensive Cancer Center, University of California Irvine, Irvine, CA 92697, USA
6
Edwards Life Sciences Center for Advanced Cardiovascular Technology, University of California Irvine, Irvine, CA 92697, USA
7
Department of Biomedical Engineering, University of California Irvine, Irvine, CA 92697, USA
8
Department of Biological Chemistry, University of California Irvine, Irvine, CA 92697, USA
9
Department of Chemistry, Incheon National University, Incheon 22012, Korea
10
Research Institute of Basic Sciences, Incheon National University, Incheon 22012, Korea
*
Author to whom correspondence should be addressed.
These authors contributed equally to this work.
Sensors 2020, 20(17), 4667; https://doi.org/10.3390/s20174667
Received: 24 July 2020 / Revised: 15 August 2020 / Accepted: 17 August 2020 / Published: 19 August 2020
(This article belongs to the Special Issue Lab-on-a-Chip and Microfluidic Sensors)
Antibiotic-resistant bacteria have emerged as an imminent global threat. The lack of rapid and sensitive diagnostic techniques leaves health care providers with inadequate resources for guiding therapy and risks the lives of patients. The traditional plate culturing methods for identifying antibiotic-resistant bacteria is laborious and time-consuming. Bulk PCR (Polymerase Chain Reaction) and qPCR are limited by poor detection sensitivity, which is critical for the early-stage detection of bloodstream infections. In this study, we introduce a technique for detecting β-lactamase-producing bacteria at single-cell sensitivity based on a commercial β-lactamase sensor (Fluorocillin), droplet microfluidics, and a custom 3D particle counter. Bacteria-containing samples were encapsulated within picoliter-sized droplets at the single-cell level and cultured within water-in-oil droplets containing antibiotics and the Fluorocillin sensor. Then, fluorescent droplets were digitally quantified with the 3D particle counter, which is capable of analyzing milliliter-scale volumes of collected droplets within 10 min. The fluorescence signal from single-colony droplets was detectable in less than 5 h, and the 3D scanning was performed in less than 10 min, which was significantly faster than conventional culture-based methods. In this approach, the limit of detection achieved was about 10 bacterial cells per mL of sample, and the turnaround time from sample to result was less than 6 h. This study demonstrates a promising strategy for the detection of β-lactamase-producing bacteria using the recently developed IC 3D system. View Full-Text
Keywords: IC 3D; droplet microfluidics; digital quantification IC 3D; droplet microfluidics; digital quantification
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MDPI and ACS Style

Li, Y.; Cherukury, H.; Labanieh, L.; Zhao, W.; Kang, D.-K. Rapid Detection of β-Lactamase-Producing Bacteria Using the Integrated Comprehensive Droplet Digital Detection (IC 3D) System. Sensors 2020, 20, 4667.

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