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An aptamer based fluorometric microcystin-LR assay using DNA strand-based competitive displacement.
Chinnappan, Raja; AlZabn, Razan; Abu-Salah, Khalid M; Zourob, Mohammed.
Afiliación
  • Chinnappan R; Department of Chemistry, Alfaisal University, Al Zahrawi Street, Al Maather, Al Takhassusi Rd, Riyadh, 11533, Saudi Arabia.
  • AlZabn R; Department of Chemistry, Alfaisal University, Al Zahrawi Street, Al Maather, Al Takhassusi Rd, Riyadh, 11533, Saudi Arabia.
  • Abu-Salah KM; Department of Nanomedicine, King Abdullah International Medical Research Center, Ministry of the National Guard-Health Affairs, Riyadh/ King Saud bin Abdulaziz University for Health Sciences, Riyadh, Saudi Arabia.
  • Zourob M; Department of Chemistry, Alfaisal University, Al Zahrawi Street, Al Maather, Al Takhassusi Rd, Riyadh, 11533, Saudi Arabia. mzourob@alfaisal.edu.
Mikrochim Acta ; 186(7): 435, 2019 06 13.
Article en En | MEDLINE | ID: mdl-31197617
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.
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Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Espectrometría de Fluorescencia / ADN / Técnicas Biosensibles / Aptámeros de Nucleótidos / Microcistinas Idioma: En Revista: Mikrochim Acta Año: 2019 Tipo del documento: Article País de afiliación: Arabia Saudita Pais de publicación: Austria
Texto completo
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Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Espectrometría de Fluorescencia / ADN / Técnicas Biosensibles / Aptámeros de Nucleótidos / Microcistinas Idioma: En Revista: Mikrochim Acta Año: 2019 Tipo del documento: Article País de afiliación: Arabia Saudita Pais de publicación: Austria