Aptamer cocktails: Enhancement of sensing signals compared to single use of aptamers for detection of bacteria

Abstract

Microbial cells have many binding moieties on their surface for binding to their specific bioreceptors. The whole-cell SELEX process enables the isolation of various aptamers that can bind to different components on the cell surface such as proteins, polysaccharides, or flagella with high affinity and specificity. Here, we examine the binding capacity of an aptamer mixture (aptamer cocktail) composed of various combinations of 3 different DNA aptamers isolated from Escherichia coli and compare it with one of the single aptamers using fluorescence-tagged aptamers. The aptamer mixtures showed higher fluorescence signal than did any single aptamer used, which suggests that use of aptamer mixtures can enhance the sensitivity of detection of microbial cells. To further evaluate this effect, the signal enhancement and improvement of sensitivity provided by combinatorial use of aptamers were examined in an electrochemical detection system. With regard to current decreases, the aptamer cocktail immobilized on gold electrodes performed better than a single aptamer immobilized on gold electrodes did. Consequently, the detection limit achieved using the aptamers individually was approximately 18 times that when the 3 aptamers were used in combination. These results support the use of aptamer cocktails for detection of complex targets such as E. coli with enhanced sensitivity. (C) 2013 Elsevier B.V. All rights reserved.