Details

Autor(en) / Beteiligte

• Chinnappan, Raja
• AlZabn, Razan
• Abu-Salah, Khalid M.
• Zourob, Mohammed

Titel

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

Ist Teil von

• Mikrochimica acta (1966), 2019-07, Vol.186 (7), p.435-435, Article 435

Ort / Verlag

Vienna: Springer Vienna

Erscheinungsjahr

2019

Quelle

MEDLINE

Beschreibungen/Notizen

• 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.