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

Int J Biol Macromol. 2019 May 15:129:91-97. doi: 10.1016/j.ijbiomac.2019.02.030. Epub 2019 Feb 6.

Abstract

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 μM for 9S aptamer, while 13S aptamer enabled a wider linear detection range between 5 pM and 500 μM. 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.

Keywords: DNA aptamer; GO-SELEX; Sarcosine.

MeSH terms

  • Aptamers, Nucleotide* / chemistry
  • Biological Assay* / methods
  • Biological Assay* / standards
  • Biosensing Techniques* / methods
  • Biosensing Techniques* / standards
  • Fluorescence*
  • Humans
  • Nucleic Acid Conformation
  • SELEX Aptamer Technique
  • Sarcosine / analysis*
  • Sensitivity and Specificity

Substances

  • Aptamers, Nucleotide
  • Sarcosine