A highly sensitive DNA aptamer-based fluorescence assay for sarcosine detection down to picomolar levels
Makaleler > Tam Makale · 2019
Yazarlar: Canan Özyurt, Zeynep Çelik Canbay, Erhan DİNÇKAYA, Serap EVRAN
International Journal of Biological Macromolecules, SCI Expanded İndekslerine Giren Dergi, Uluslararası, Wos, 2019, cilt 129, ss 91-97
DOI: 10.1016/j.ijbiomac.2019.02.030
Özet
Sarcosine is an amino acid derivative, which is considered as a key metabolite in various metabolic processes. Therefore, simple and sensitive detection methods are needed for further understanding its metabolic role and diagnostic value. In this study, we developed a novel method that meets the need for practical and sensitive detection in a complex medium mimicking urine conditions. For this aim, we selected sarcosine-specific DNA aptamers using graphene oxide-assisted systemic evolution of ligands by exponential enrichment (GO-SELEX). The candidate aptamers were labeled with 6-carboxyfluorescein (6-FAM) at their 5' ends. Two aptamers, namely 9S and 13S produced a significant fluorescence signal upon sarcosine binding. Both aptamers enabled a sensitive analysis with a detection limit of 0.5 pM. The linear detection ranged between 5 pM and 50 mu M for 9S aptamer, while 13S aptamer enabled a wider linear detection range between 5 pM and 500 mu M. The aptamer-based assay allowed rapid detection with no need for chemical derivatization of sarcosine and sophisticated instruments. Moreover, the aptamer-based assay was free of interference from urea and human serum albumin. (C) 2019 Elsevier B.V. All rights reserved.