title: G-quadruplex aptamer targeting Protein A and its capability to detect Staphylococcus aureus demonstrated by ELONA.
abstract: Aptamers for whole cell detection are selected mostly by the Cell-SELEX procedure. Alternatively, the use of specific cell surface epitopes as target during aptamer selections allows the development of aptamers with ability to bind whole cells. In this study, we integrated a formerly selected Protein A-binding aptamer PA#2/8 in an assay format called ELONA (Enzyme-Linked OligoNucleotide Assay) and evaluated the ability of the aptamer to recognise and bind to Staphylococcus aureus presenting Protein A on the cell surface. The full-length aptamer and one of its truncated variants could be demonstrated to specifically bind to Protein A-expressing intact cells of S. aureus, and thus have the potential to expand the portfolio of aptamers that can act as an analytical agent for the specific recognition and rapid detection of the bacterial pathogen. The functionality of the aptamer was found to be based on a very complex, but also highly variable structure. Two structural key elements were identified. The aptamer sequence contains several G-clusters allowing folding into a G-quadruplex structure with the potential of dimeric and multimeric assembly. An inverted repeat able to form an imperfect stem-loop at the 5'-end also contributes essentially to the aptameric function.
authors.affiliation: UFZ - Helmholtz Centre for Environmental Research, Department of Soil Ecology, Theodor-Lieser-Straße 4, 06120 Halle, Germany.
authors.forename: Regina
authors.initials: R
authors.lastname: Stoltenburg
authors.affiliation: P.J.Šafárik University, Faculty of Sciences, Institute of Chemistry, Dept. of Biochemistry, Moyzesova 11, 04011 Košice, Slovakia.
authors.forename: Petra
authors.initials: P
authors.lastname: Krafčiková
authors.affiliation: P.J.Šafárik University, Faculty of Sciences, Institute of Chemistry, Dept. of Biochemistry, Moyzesova 11, 04011 Košice, Slovakia.
authors.forename: Viktor
authors.initials: V
authors.lastname: Víglaský
authors.affiliation: UFZ - Helmholtz Centre for Environmental Research, Department Environmental and Biotechnology Centre, Permoserstraße 15, 04318 Leipzig, Germany.
authors.forename: Beate
authors.initials: B
authors.lastname: Strehlitz
mesh_terms: D052157:Aptamers, Nucleotide / Q000737:chemistry*; D054856:G-Quadruplexes; D013205:Staphylococcal Protein A / Q000032:analysis*; D013211:Staphylococcus aureus
chemical_list: D052157:Aptamers, Nucleotide; D013205:Staphylococcal Protein A