A highly sensitive DNA aptamer-based fluorescence assay for sarcosine detection down to picomolar levels


Özyurt C., Canbay Z. Ç. , DİNÇKAYA E. , EVRAN S.

INTERNATIONAL JOURNAL OF BIOLOGICAL MACROMOLECULES, cilt.129, ss.91-97, 2019 (SCI İndekslerine Giren Dergi) identifier identifier identifier

  • Cilt numarası: 129
  • Basım Tarihi: 2019
  • Doi Numarası: 10.1016/j.ijbiomac.2019.02.030
  • Dergi Adı: INTERNATIONAL JOURNAL OF BIOLOGICAL MACROMOLECULES
  • Sayfa Sayıları: ss.91-97

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.