Separation, Identification, and Characterization of Microorganisms by Capillary Electrophoresis

Abstract

The use of capillary electrophoresis (CE) for the analysis, identification, and characterization of microorganisms has been gaining in popularity. The advantages of CE, such as small sample requirements, minimal sample preparation, rapid and simultaneous analysis, ease of quantitation and identification, and viability assessment, make it an attractive technique for the analysis of microbial analytes. As this instrumental method has evolved, higher peak efficiencies have been achieved by optimizing CE conditions, such as pH, ionic strength, and polymer additive concentration. Experimental improvements have allowed better quantitation and more accurate results. Many practical applications of this technique have been investigated. Viability and identification of microbes can be accomplished in a single analysis. This is useful for evaluation of microbial analytes in consumer products. Diagnosis of microbe-based diseases is now possible, in some cases, without the need for culture methods. Microbe-molecule, virus-antibody, or bacteria-antibiotic interactions can be monitored using CE, allowing for the screening of possible drug candidates. Fermentation can be monitored using this system. This instrumental approach can be adapted to many different applications, including assessing the viability of sperm cells. Progress has been made in the development of microelectrophoresis instrumentation. These advances will eventually allow the development of small, dedicated devices for the rapid, repetitive analyses of specific microbial samples. Although these methods may never fully replace traditional approaches, they are proving to be a valuable addition to the collection of techniques used to analyze, quantitate, and characterize microbes. This review outlines the recent developments in this rapidly growing field

Separation of Plant Pathogens from Different Hosts and Tissues by Capillary Electromigration Techniques

“…CIEF. The previously published results 6,7,[9][10][11]19,23,33,34,39,40 indicate that the isoelectric points of MOs could belong to their identification attributes in the future. However, only a small number of pIs of MOs have been tabulated until now.…”

Separation of Plant Pathogens from Different Hosts and Tissues by Capillary Electromigration Techniques

“…CIEF. The previously published results 6,7,[9][10][11]19,23,33,34,39,40 indicate that the isoelectric points of MOs could belong to their identification attributes in the future. However, only a small number of pIs of MOs have been tabulated until now.…”

On-Column Labeling of Gram-Positive Bacteria with a Boronic Acid Functionalized Squarylium Cyanine Dye for Analysis by Polymer-Enhanced Capillary Transient Isotachophoresis

“…Dilute PEO has been employed as a buffer additive in various CE studies to focus bacteria samples. − ,,,, The multiple (sharp) peaks for Bg spores spread throughout a detection window of 2–3 min (as shown in Figure a) were, indeed, focused into a more narrow (approximately 30 s) detection zone by the addition of 0.0125% of PEO 600 000 to the separation buffer (Figure b–d). Although a single, sharp peak for Bg spores was obtained in some runs with added PEO (Figure b), these CE-LIF conditions demonstrated low reproducibility with multiple peaks for Bg spore samples still appearing in some replicate runs employing PEO.…”

Combined capillary electrophoresis and DNA‐fluorescence in situ hybridization for rapid molecular identification of Salmonella Typhimurium in mixed culture

“…Alternatively, our CE-FISH approach could also be adapted for use at the microscale, in miniaturized, "lab on a chip" applications [4,29]. Such an approach may be especially attractive for single cell detection of microbial pathogens [3,30].…”

Separation of Plant Pathogens from Different Hosts and Tissues by Capillary Electromigration Techniques

“…CIEF. The previously published results 6,7,[9][10][11]19,23,33,34,39,40 indicate that the isoelectric points of MOs could belong to their identification attributes in the future. However, only a small number of pIs of MOs have been tabulated until now.…”

On-Column Labeling of Gram-Positive Bacteria with a Boronic Acid Functionalized Squarylium Cyanine Dye for Analysis by Polymer-Enhanced Capillary Transient Isotachophoresis

“…Dilute PEO has been employed as a buffer additive in various CE studies to focus bacteria samples. − ,,,, The multiple (sharp) peaks for Bg spores spread throughout a detection window of 2–3 min (as shown in Figure a) were, indeed, focused into a more narrow (approximately 30 s) detection zone by the addition of 0.0125% of PEO 600 000 to the separation buffer (Figure b–d). Although a single, sharp peak for Bg spores was obtained in some runs with added PEO (Figure b), these CE-LIF conditions demonstrated low reproducibility with multiple peaks for Bg spore samples still appearing in some replicate runs employing PEO.…”

Combined capillary electrophoresis and DNA‐fluorescence in situ hybridization for rapid molecular identification of Salmonella Typhimurium in mixed culture

“…Alternatively, our CE-FISH approach could also be adapted for use at the microscale, in miniaturized, "lab on a chip" applications [4,29]. Such an approach may be especially attractive for single cell detection of microbial pathogens [3,30].…”

Separation of Plant Pathogens from Different Hosts and Tissues by Capillary Electromigration Techniques

“…CIEF. The previously published results 6,7,[9][10][11]19,23,33,34,39,40 indicate that the isoelectric points of MOs could belong to their identification attributes in the future. However, only a small number of pIs of MOs have been tabulated until now.…”

On-Column Labeling of Gram-Positive Bacteria with a Boronic Acid Functionalized Squarylium Cyanine Dye for Analysis by Polymer-Enhanced Capillary Transient Isotachophoresis

“…Dilute PEO has been employed as a buffer additive in various CE studies to focus bacteria samples. − ,,,, The multiple (sharp) peaks for Bg spores spread throughout a detection window of 2–3 min (as shown in Figure a) were, indeed, focused into a more narrow (approximately 30 s) detection zone by the addition of 0.0125% of PEO 600 000 to the separation buffer (Figure b–d). Although a single, sharp peak for Bg spores was obtained in some runs with added PEO (Figure b), these CE-LIF conditions demonstrated low reproducibility with multiple peaks for Bg spore samples still appearing in some replicate runs employing PEO.…”

Combined capillary electrophoresis and DNA‐fluorescence in situ hybridization for rapid molecular identification of Salmonella Typhimurium in mixed culture

“…Alternatively, our CE-FISH approach could also be adapted for use at the microscale, in miniaturized, "lab on a chip" applications [4,29]. Such an approach may be especially attractive for single cell detection of microbial pathogens [3,30].…”