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Microchimica Acta

, 186:435 | Cite as

An aptamer based fluorometric microcystin-LR assay using DNA strand-based competitive displacement

  • Raja Chinnappan
  • Razan AlZabn
  • Khalid M. Abu-Salah
  • Mohammed ZourobEmail author
Original Paper

Abstract

The high-affinity region of a truncated aptamer was applied to the development of a sensitive method for the determination of microcystin-LR (MC-LR) using competitive displacement and molecular beacons. In this assay, the fluorophore and quencher labelled complementary sequences of the aptamer are hybridized with the truncated aptamer to form a fluorophore-quencher pair. In the presence of MC-LR, the aptamer duplex dissociates, and the fluorophore-quencher pair is separated. This turn leads to an increase in the yellow fluorescence which is best measured at excitation/emission wavelengths of 555/580 nm. One of the truncated aptamers showed a 50-fold increase in the affinity (0.93 nM) compared to the wild type aptamer (50 nM). The truncated sequence shows considerable cross-reactivity with L congeners but none with other congeners. The assay works in 0.5 to 200 nM MC-LR concentration range. It was applied to spiked tap water samples and gave recoveries around 95 ± 5%.

Graphical abstract

Schematic representation of a method for determination of microcystin-LR via fluorescence that is induced by competitive displacement of complementary DNA strands in a truncated dsDNA aptamer.

Keywords

Aptasensor SELEX Fluorescence assay Marine toxins Microcystin-LR Cyanobacterial toxins Aptamer truncation Blue-green algae Harmful algae Graphene oxide 

Notes

Compliance with ethical standards

The author(s) declare that they have no competing interests.

Supplementary material

604_2019_3504_MOESM1_ESM.docx (40 kb)
ESM 1 (DOCX 40 kb)

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Copyright information

© Springer-Verlag GmbH Austria, part of Springer Nature 2019

Authors and Affiliations

  • Raja Chinnappan
    • 1
  • Razan AlZabn
    • 1
  • Khalid M. Abu-Salah
    • 2
  • Mohammed Zourob
    • 1
    • 3
    Email author
  1. 1.Department of ChemistryAlfaisal UniversityRiyadhSaudi Arabia
  2. 2.Department of Nanomedicine, King Abdullah International Medical Research Center, Ministry of the National Guard-Health AffairsRiyadh/ King Saud bin Abdulaziz University for Health SciencesRiyadhSaudi Arabia
  3. 3.King Faisal Specialist Hospital and Research CenterRiyadhSaudi Arabia

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