Abstract
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.
Highlights
Due to the use of β-lactam antibiotics in medicine, agriculture, and veterinary medicine, bacteria have evolved to survive in the presence of antibiotics by mutating and encoding extended-spectrumSensors 2020, 20, 4667; doi:10.3390/s20174667 www.mdpi.com/journal/sensorsSensors 2020, 20, 4667 β-lactamases (ESBLs)
This study demonstrates a promising strategy for the detection of β-lactamase-producing bacteria using the recently developed IC 3D system
We recently reported a technology termed Integrated Comprehensive Droplet Digital Detection (IC 3D), which is capable of analyzing milliliters of droplets within minutes to identify target pathogens
Summary
Due to the use of β-lactam antibiotics in medicine, agriculture, and veterinary medicine, bacteria have evolved to survive in the presence of antibiotics by mutating and encoding extended-spectrumSensors 2020, 20, 4667; doi:10.3390/s20174667 www.mdpi.com/journal/sensorsSensors 2020, 20, 4667 β-lactamases (ESBLs). BSI can lead to sepsis and result in a very high mortality rate. 1 million individuals are hospitalized due to sepsis annually in the US alone with at least 30,000 of those patients dying in the hospital, and the numbers are rising every year [2]. The high mortality rate of sepsis can be attributed to the lack of rapid diagnostics tools to direct the proper administration of antibiotics by healthcare providers. Until the results of the culture are confirmed, physicians are generally directed to administer a cocktail of broad-spectrum antibiotics in the hopes that one or more of the antibiotics will be effective in curbing the infection [4,5]
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