Antibiotic susceptibility test based on the dielectrophoretic behavior of elongated <i>Escherichia coli</i> with cephalexin treatment

Chung, Ching-Che; Cheng, I-Fang; Yang, Wen-Horng; Chang, Hsien-Chang

doi:10.1063/1.3600650

Cited by 29 publications

(31 citation statements)

References 20 publications

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“…Heat-killed cells were used as a model system to study DEP response in most cases [9][10][11][12]. Since extensive heat could change the electric properties of cell wall and cytoplasm, the DEP behavior of cells could differ greatly between heat-and antibiotic-inactivated bacterial cells.…”

Section: Introductionmentioning

confidence: 99%

Dielectrophoretic characterization of antibiotic-treated Mycobacterium tuberculosis complex cells

et al. 2015

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Multi-drug resistant tuberculosis (MDR-TB) has become a serious concern for proper treatment of patients. As a phenotypic method, dielectrophoresis can be useful but is yet to be attempted to evaluate Mycobacterium tuberculosis complex cells. This paper investigates the dielectrophoretic behavior of Mycobacterium bovis (Bacillus Calmette-Guérin, BCG) cells that are treated with heat or antibiotics rifampin (RIF) or isoniazid (INH). The experimental parameters are designed on the basis of our sensitivity analysis. The medium conductivity (σ(m)) and the frequency (f) for a crossover frequency (f(xo1)) test are decided to detect the change of σ(m)-f(xo1) in conjunction with the drug mechanism. Statistical modeling is conducted to estimate the distributions of viable and nonviable cells from the discrete measurement of f (xo1). Finally, the parameters of the electrophysiology of BCG cells, C(envelope) and σ(cyto), are extracted through a sampling algorithm. This is the first evaluation of the dielectrophoresis (DEP) approach as a means to assess the effects of antimicrobial drugs on M. tuberculosis complex cells.

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Section: Introductionmentioning

confidence: 99%

Dielectrophoretic characterization of antibiotic-treated Mycobacterium tuberculosis complex cells

et al. 2015

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“…(1b), (3), (6), and (7) can be replaced by its effective value, 4,20 which takes into account the additional surface conductance associated with the double layer effect outside the particle (cell). Equation (3) or (7) shows a linear relationship between f pk R or f co R and r 1 .…”

Section: Theory a Current Theorymentioning

confidence: 99%

“…Dielectrophoresis (DEP) and electrorotation (ER) were employed extensively in particle=cell manipulation [1][2][3] and characterization [4][5][6][7][8] in the literature. DEP (or ER) is a phenomenon associated with the movement (or rotation) of a particle or cell under the action of a non-uniform (or uniformly rotating) electric field.…”

Section: Introductionmentioning

confidence: 99%

Refinement of the theory for extracting cell dielectric properties from dielectrophoresis and electrorotation experiments

2011

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A modified theory is proposed for extracting cell dielectric properties from the peak frequency measurement of electrorotation (ER) and the crossover frequency measurement of dielectrophoresis (DEP). Current theory in the literature is based on the low frequency (DC) approximations for the equivalent cell permittivity and conductivity, which are valid when the measurements are performed in a medium with conductivity less than 1 mS=m. The present theory extracts the cell properties through optimizing an expression for the medium conductivity in terms of the peak ER, or DEP crossover, frequency according to its definition using full expressions of equivalent cell permittivity and conductivity. Various levels of approximation of the theory are proposed and discussed through a scaling analysis. The present theory can extract both membrane and interior properties from the low and the high peak ER, or DEP crossover, frequencies for any medium conductivity provided the peak ER, or DEP crossover, frequency exists. It can be reduced to the linear theory for the low peak ER and DEP crossover frequencies in the literature when the medium conductivity is less than 10 mS=m. However, we can determine the membrane capacitance and conductance via the slope and intercept, respectively, of the straight line fitting of the ER peak and DEP frequency against medium conductivity data according to the linear theory only when the intercept dominates the experimental uncertainty, which occurs when the medium conductivity is less than 1 mS=m in practice.

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“…Bacteria has also been separated from other bacteria based on differing genera [61,62,71,72,75,76], species [57], and serotypes [77]. Other researchers have used DEP as a means to separate or identify bacteria according to viability [68,78,79] and antibiotic resistance [80][81][82][83][84]. In some cases, DEP has been used to concentrate bacteria to a specific area for subsequent analysis or identification [60,61,71,[85][86][87].…”

Section: Raman Studies Of Mycobacteriummentioning

confidence: 99%

The use of microfluidics and dielectrophoresis for separation, concentration, and identification of bacteria

et al. 2016

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Traditional bacterial analyses take one to two days under favorable conditions where the bulk of the time is spent waiting for bacteria to divide and grow until visual colonies can be observed for identification. In the case of bacteria with slow doubling times, this process can take weeks. This delay in analysis is unacceptable, especially in cases of life threatening diseases or emergencies. It is clear that in order to decrease the analysis time of the bacteria, the culturing and growth step must be circumvented. The goal of this research is to design, build, and test a device that could decrease the analysis time of bacteria using label-free methods of dielectrophoresis and Raman spectroscopy.Testing for device design was performed with clinical samples in mind, which consist of bacteria grown in a variety of environmental conditions (i.e. available food sources, growth stage, temperature, etc.) and accompanied by sample debris. Raman spectra of bacteria grown in varying media and metabolic stages were collected and analyzed. Results indicate that growth phase and media have an impact on Raman spectra iv and is distinguishable by linear discriminant analysis (LDA). Despite these spectral differences, it was found that LDA classification of closely related bacteria remains fairly high (90%) regardless of growth phase. Sample debris were also considered in device design and accommodated for by dielectrophoresis. Devices were built with the goal to isolate bacteria from a mixed sample and simultaneously acquire Raman spectra for identification.For this dissertation, a device was designed, built, and tested that incorporates dielectrophoresis for particle isolation and Raman spectroscopy for identification. The device was modeled in COMSOL to ensure that an appropriate electrical field gradient could be obtained to isolate bacteria from 5 µm diameter polystyrene spheres. The device was built and successfully trapped bacteria away from polystyrene spheres and Raman spectra of the bacteria were collected while trapped. These results indicate a clear potential for contactless dielectrophoresis-Raman devices to isolate and identify bacteria from sample debris, and thereby decrease the analysis time of bacteria. Typical bacterial analysis involves culturing and visualizing colonies on an array of agar plates. The growth patterns and colors among the array are used to identify the bacteria. For fast growing bacteria such as Escherichia coli, analysis will take one to two days. However, slow growing bacteria such as mycobacteria can take weeks to identify.In addition, there are some species of bacteria that are viable but nonculturable. This lengthy analysis time is unacceptable for life-threatening infections and emergency situations. It is clear that to decrease the analysis of the bacteria, the culturing and growth steps must be avoided. The goal of this research is to design, build, and test a device that could decrease the analysis time of bacteria.

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Antibiotic susceptibility test based on the dielectrophoretic behavior of elongated Escherichia coli with cephalexin treatment

Cited by 29 publications

References 20 publications

Dielectrophoretic characterization of antibiotic-treated Mycobacterium tuberculosis complex cells

Dielectrophoretic characterization of antibiotic-treated Mycobacterium tuberculosis complex cells

Refinement of the theory for extracting cell dielectric properties from dielectrophoresis and electrorotation experiments

The use of microfluidics and dielectrophoresis for separation, concentration, and identification of bacteria

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