Microfluidics-Based Lab-on-Chip Systems in DNA-Based Biosensing: An Overview

Dutse, Sabo Wada; Yusof, Nor Azah

doi:10.3390/s110605754

Cited by 101 publications

(50 citation statements)

References 72 publications

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“…To date, various methods have been developed and introduced in detecting pathogenic diseases but electrochemical DNA detection methods have now become the focal point of consideration because of specificity, rapid analysis time, and low cost (Palecek et al 1998). DNA biosensors respond to a measurable signal that is proportional to the concentration of complementary (target) DNA sequence (Dutse and Yusof 2011;Dutse et al 2012). Moreover, the concentration of genetic targets in biological samples is low and these compounds are suitable for determination by a DNA biosensor.…”

Section: Introductionmentioning

confidence: 99%

An Electrochemical Biosensor for the Determination ofGanoderma boninensePathogen Based on a Novel Modified Gold Nanocomposite Film Electrode

et al. 2014

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A sensitive approach for the determination of Ganoderma boninense DNA is reported based on an electrochemical affinity system using a modified gold sensor. Covalent attachment of probe DNA was achieved by attachment of the amine group to a carboxylic acid group of a 3,3'-dithiodipropionic acid monolayer on a nanocomposite film of gold nanoparticles bound to poly(3,4-ethylenedioxythiophen)-poly(styrenesulfonate) on a gold working electrode. The electrochemical detection of sequence-specific DNA of probe and target DNA hybridization was monitored using a new ruthenium complex [Ru(dppz) 2 (qtpy)Cl 2 ; dppz = dipyrido [3,2-a:2',3'-c] phenazine; qtpy = 2,2',-4,4".4'4"'-quarterpyridyl redox marker. The potential was selected through the study of the electrochemical behavior of trisaminomethane-hydrochloride containing a ethylenediaminetetraacetic acid supporting electrolyte on the bare and modified gold electrode. The effect of the hybridization temperature and time were measured. The sensor demonstrated specific detection for the target over a concentration range of 1.0 × 10 −15 M to 1.0 × 10 −9 M with a detection limit of 1.59 × 10 −17 M. Control experiments verified the specificity of the biosensor in the presence of a single mismatched DNA sequence. This detection technology was shown to be effective in terms of sensitivity and selectivity of hybridization events and is a promising device for early detection of Ganoderma boninense and other pathogenic threat agents.

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Section: Introductionmentioning

confidence: 99%

An Electrochemical Biosensor for the Determination ofGanoderma boninensePathogen Based on a Novel Modified Gold Nanocomposite Film Electrode

et al. 2014

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“…Thirdly, sensors can be integrated within microfluidic devices to enable label-free detection. Lastly, these devices are inexpensive to produce and often require minimal operator time and skill (Craighead, 2006;Dittrich and Manz, 2006;Dutse and Yusof, 2011;Jiang et al, 2011).…”

Section: Introductionmentioning

confidence: 99%

A controlled microfluidic electrochemical lab-on-a-chip for label-free diffusion-restricted DNA hybridization analysis

Ben‐Yoav

Dykstra

Bentley

et al. 2015

Biosensors and Bioelectronics

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“…This information can be coupled to risk management of the unwanted growth of bacteria in food and hence help the food industry combat food spoilage. An attractive platform for implementing such an assay would be a microfluidics-based lab-on-chip system (30). Such a platform allows for rapid measurement of pH i values under dynamically changing conditions (change of medium types or supplements) coupled to monitoring of the dynamics of spore germination and outgrowth.…”

Section: Discussionmentioning

confidence: 99%

Intracellular pH Response to Weak Acid Stress in Individual Vegetative Bacillus subtilis Cells

Pandey

Vischer

Smelt

et al. 2016

Appl Environ Microbiol

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Intracellular pH (pH i ) critically affects bacterial cell physiology. Hence, a variety of food preservation strategies are aimed at perturbing pH i homeostasis. Unfortunately, accurate pH i quantification with existing methods is suboptimal, since measurements are averages across populations of cells, not taking into account interindividual heterogeneity. Yet, physiological heterogeneity in isogenic populations is well known to be responsible for differences in growth and division kinetics of cells in response to external stressors. To assess in this context the behavior of intracellular acidity, we have developed a robust method to quantify pH i at single-cell levels in Bacillus subtilis. Bacilli spoil food, cause disease, and are well known for their ability to form highly stress-resistant spores. Using an improved version of the genetically encoded ratiometric pHluorin (IpHluorin), we have quantified pH i in individual B. subtilis cells, cultured at an external pH of 6.4, in the absence or presence of weak acid stresses. In the presence of 3 mM potassium sorbate, a decrease in pH i and an increase in the generation time of growing cells were observed. Similar effects were observed when cells were stressed with 25 mM potassium acetate. Time-resolved analysis of individual bacteria in growing colonies shows that after a transient pH decrease, long-term pH evolution is highly cell dependent. The heterogeneity at the single-cell level shows the existence of subpopulations that might be more resistant and contribute to population survival. Our approach contributes to an understanding of pH i regulation in individual bacteria and may help scrutinizing effects of existing and novel food preservation strategies. IMPORTANCEThis study shows how the physiological response to commonly used weak organic acid food preservatives, such as sorbic and acetic acids, can be measured at the single-cell level. These data are key to coupling often-observed single-cell heterogeneous growth behavior upon the addition of weak organic acid food preservatives. Generally, these data are gathered in the form of plate counting of samples incubated with the acids. Here, we visualize the underlying heterogeneity in cellular pH homeostasis, opening up avenues for mechanistic analyses of the heterogeneity in the weak acid stress response. Thus, microbial risk assessment can become more robust, widening the scope of use of these well-known weak organic acid food preservatives.

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Microfluidics-Based Lab-on-Chip Systems in DNA-Based Biosensing: An Overview

Cited by 101 publications

References 72 publications

An Electrochemical Biosensor for the Determination ofGanoderma boninensePathogen Based on a Novel Modified Gold Nanocomposite Film Electrode

An Electrochemical Biosensor for the Determination ofGanoderma boninensePathogen Based on a Novel Modified Gold Nanocomposite Film Electrode

A controlled microfluidic electrochemical lab-on-a-chip for label-free diffusion-restricted DNA hybridization analysis

Intracellular pH Response to Weak Acid Stress in Individual Vegetative Bacillus subtilis Cells

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