DEVELOPMENT of DNA APTAMER-BASED METHOD FOR DETECTION of ASYMMETRIC DIMETHYLARGININE


Bora B. , Evran S. , Özyurt C., Bilici M.

IV. Turkey In Vitro Diagnostic Symposia, İzmir, Turkey, 20 - 22 February 2019, vol.44, pp.42

  • Publication Type: Conference Paper / Full Text
  • Volume: 44
  • City: İzmir
  • Country: Turkey
  • Page Numbers: pp.42

Abstract

DEVELOPMENT of DNA APTAMER-BASED METHOD FOR DETECTION of ASYMMETRIC DIMETHYLARGININE Merve Bilici, Serap Evran, Canan Özyurt, Burhan Bora Ege University, Biochemistry, Izmir, Turkey Objectives: In recent studies, it has been shown that there is a relationship between asymmetric dimethylarginine (ADMA) levels and many diseases, such as cardiovascular diseases. Detection of ADMA becomes important as a new approach in diagnosis, treatment and treatment of these diseases. Due to the disadvantages of current ADMA detection methods and limited use in routine analysis, there is an increasing need for new methods. Reliable, specific, fast and costeffective test kits are needed for detection of ADMA, which is considered as a cardiovascular risk biomarker. In this study, we aimed to select ADMA-specific DNA aptamers and to develop a new detection method. Materials-Methods: Graphene Oxide (GO) - SELEX method was used to develop ADMA aptamers. In the final round of SELEX, the enriched oligonucleotide library was cloned into pTZ57R / T plasmid. Sanger sequence analysis was performed and the selected aptamer was synthesized with FAM-label. The FAM-labeled aptamer was incubated with increasing concentrations of ADMA and the change in fluorescence signal was monitored. Results: 13 SELEX rounds were performed for selection of ADMA aptamers. Aptamers from the final SELEX round were analyzed in terms of secondary structure and conserved sequence. The parameters, such as detection time, detection limit, linear range and specificity of the method was compared with commercial ELISA kits. Conclusions: A novel fluorimetric detection method was developed by using ADMA-specific DNA aptamer. Keywords: Asymmetric dimethylarginine (ADMA), Aptamer, SELEX