Real-time aptamer quantum dot fluorescent flow sensor.

The goal of this work was to develop and test a novel real-time biosensing approach which can be adapted to either environmental or clinical monitoring of biological pathogens. We have developed a working prototype of a real-time aptamer-based fluorescent flow sensor. The sensor utilizes a competitive displacement approach to measure the binding of the analyte, which keeps the nonspecific binding below detectable levels. The complex of surface-immobilized DNA aptamer with fluorescent complementary oligonucleotide releases the oligonucleotide upon binding with a specific target, which is translated by a decrease in fluorescence. Bright and stable fluorescence of quantum dots is utilized for prolonged detection of the analyte in flow conditions. The real-time sensor prototype is developed with previously characterized ATP-specific aptamer and is capable of specifically detecting 0.1 mM of ATP in biological buffer, with a quantitative response up to 5 mM. The developed prototype is portable and easy to use and its design allows further miniaturization and multiplexing. The developed real-time sensing approach can be adapted to a variety of targets of environmental and clinical significance.