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

被引:23
|
作者
Ozyurt, Canan [1 ]
Canbay, Zeynep Celik [1 ]
Dinckaya, Erhan [1 ]
Evran, Serap [1 ]
机构
[1] Ege Univ, Fac Sci, Dept Biochem, TR-35100 Bornova, Turkey
关键词
DNA aptamer; GO-SELEX; Sarcosine; SELECTION; METABOLISM; SAMPLES; SENSOR;
D O I
10.1016/j.ijbiomac.2019.02.030
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
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.
引用
收藏
页码:91 / 97
页数:7
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