Journal
TALANTA
Volume 106, Issue -, Pages 309-314Publisher
ELSEVIER SCIENCE BV
DOI: 10.1016/j.talanta.2012.11.018
Keywords
Enzyme linked aptamer assay; Colorimetry; Cocaine; Small molecule; Biomacromolecule
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Funding
- National Natural Science Foundation of China [20805002, 30890142, 20975007]
- Scientific Research Foundation for the Returned Overseas Chinese Scholars, MOE. China
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Enzyme linked aptamer assay (ELAA) uses an aptamer as recognition element and enzyme as signal readout element for establishing different kinds of aptasensors. We reported herein a high-throughput colorimetric aptasensor based on ELAA only requiring a single aptamer sequence for cocaine detection. An anti-cocaine aptamer was cleaved into two fragments, one of which was immobilized on a DNA-BIND 96-well plate via 5'-labeled primary amine and the other one was biotin labeled. The presence of two aptamer fragments and the target molecule led to the formation of aptamer fragments/target complexes. Streptavidin-horseradish peroxidase (SA-HRP) was used to react with biotin in order to obtain quantitative signals. A linear response towards cocaine concentration in the range of 5-200 mu M and a detection limit down to 2.8 mu M (S/N=3) were achieved. The specificity and application in real sample were validated. Furthermore, a verification test of thrombin detection in the same strategy illustrated its feasibility for not only small molecule but also biomacromolecule. With the advantage of high-throughput, easy operation, high specificity, the colorimetric assay based on ELAA requiring a single aptamer sequence opens up a new approach for detecting different kinds of targets via specific affinity recognition among target and suitably cleaved aptamer fragments. (C) 2012 Elsevier B.V. All rights reserved.
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