Capillary Electrophoresis Based Microbial Detection and Separation
In the past few decades, capillary electrophoresis (CE) has gained popularity in molecular analytical separations due to its inherent advantages over other chromatographic techniques. Recently, there has been increasing interest in applying this technique to the separation and analysis of colloidal particles including viruses, bacteria, and fungi, as there is a great need for a rapid assay of these microorganisms in various branches of the food, pharmaceutical, and medical industries. Numerous challenges still exist, however, in the development of a CE-based method for microbial analysis. The focus of this work was two-fold: 1) to develop a rapid CE based technique for the detection of Candida albicans fungi in blood samples, and 2) to physically separate multiple microorganism species using capillary isoelectric focusing. Candida albicans was successfully focused and detected from a spiked blood sample by controlling the cells’ mobility through the capillary using various surfactant detergents. A capillary isoelectric focusing method was also developed which isolated species of bacteria (i.e. Bacillus subtilus, Escherichia coli, and Pseudomonas fluorescens) into discrete zones within the capillary. Optimization of the capillary coating procedure, focusing time, and mobilization steps were also performed. Future work includes further improvements to the separation, quantitation of cells in biological samples, and developing methodology to determine the time of sample inoculation.
Faculty Mentor: Andrew Lantz, Chemistry
Ryan presented at the 239th ACS National Meeting March 20-25, 2010 in San Francisco, CA.
Page last modified January 21, 2011