A simple magnetic-assisted microfluidic method for rapid detection and phenotypic characterization of ultralow concentrations of bacteria

Abstract

Isolation and enumeration of bacteria at ultralow concentrations and antibiotic resistance profiling are of great importance for early diagnosis and treatment of bacteremia. In this work, we describe a simple, rapid, and versatile magnetic-assisted microfluidic method for rapid bacterial detection. The developed method enables magnetophoretic loading of bead-captured bacteria into the microfluidic chamber under external static and dynamic magnetic fields in 4 min. A shallow microfluidic chamber design that enables the monolayer orientation and transportation of the beads and a glass substrate with a thickness of 0.17 mm was utilized to allow high-resolution fluorescence imaging for quantitative detection. Escherichia coli (E. coli) with green fluorescent protein (GFP)-expressing gene and streptavidin-modified superparamagnetic microbeads were used as model bacteria and capturing beads, respectively. The specificity of the method was validated using Lactobacillus gasseri as a negative control group. The limit of detection and limit of quantification values were determined as 2 CFU/ml and 10 CFU/ml of E. coli, respectively. The magnetic-assisted microfluidic method is a versatile tool for the detection of ultralow concentrations of viable bacteria with the linear range of 5–5000 CFU/ml E. coli in 1 h, and providing growth curves and phenotypic characterization bead-captured E. coli in the following 5 h of incubation. Our results are promising for future rapid and sensitive antibiotic susceptibility testing of ultralow numbers of viable cells.