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Title:Ultrasensitive DNA aptasensors based on exponential amplification reaction
Author(s):Brown, Darius
Advisor(s):Lu, Yi
Department / Program:Chemistry
Degree Granting Institution:University of Illinois at Urbana-Champaign
Exponential amplification reaction (EXPAR)
Magnetic Beads
Deoxyribonucleic acid (DNA)
Abstract:This is a comprehensive project for the development of a general method to enhance the sensitivity of DNA aptasensors by coupling them with an isothermal DNA amplification technique known as exponential amplification reaction (EXPAR). The method involves two steps: (1) first, the target-induced structure-switching of DNA aptamers causes the release of trigger oligonucleotides from the aptamers immobilized on a surface; (2) then, the released trigger oligonucleotides are amplified by the EXPAR technique for ultrasensitive detection. It is shown that the detection limit of a cocaine aptasensor is improved from 5 micromolar in assays without amplification to 0.8-1micromolar when EXPAR is applied, providing a 5-fold enhancement. The flexibility in designing the sequences of the trigger oligonucleotides enables the method generally applicable to other aptasensor systems such as thrombin aptamers. By EXPAR, the method has also achieved 10-fold enhancement in sensitivity from 4nM to 400 pM for thrombin detection, in comparison with similar assays without EXPAR.
Issue Date:2014-01-16
Rights Information:Copyright 2013 Darius Z Brown
Date Available in IDEALS:2014-01-16
Date Deposited:2013-12

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