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Sensing Levofloxacin with an RNA Aptamer as a Bioreceptor.
Kramat, Janice; Kraus, Leon; Gunawan, Vincent J; Smyej, Elias; Froehlich, Philipp; Weber, Tim E; Spiehl, Dieter; Koeppl, Heinz; Blaeser, Andreas; Suess, Beatrix.
Afiliación
  • Kramat J; Synthetic RNA Biology, Department of Biology, Technical University of Darmstadt, 64287 Darmstadt, Germany.
  • Kraus L; Synthetic RNA Biology, Department of Biology, Technical University of Darmstadt, 64287 Darmstadt, Germany.
  • Gunawan VJ; Synthetic RNA Biology, Department of Biology, Technical University of Darmstadt, 64287 Darmstadt, Germany.
  • Smyej E; Synthetic RNA Biology, Department of Biology, Technical University of Darmstadt, 64287 Darmstadt, Germany.
  • Froehlich P; Self-Organizing Systems, Department of Electrical Engineering and Information Technology, Technical University of Darmstadt, 64283 Darmstadt, Germany.
  • Weber TE; Institute for BioMedical Printing Technologies, Technical University of Darmstadt, 64289 Darmstadt, Germany.
  • Spiehl D; Institute for BioMedical Printing Technologies, Technical University of Darmstadt, 64289 Darmstadt, Germany.
  • Koeppl H; Centre for Synthetic Biology, Technical University of Darmstadt, 64289 Darmstadt, Germany.
  • Blaeser A; Self-Organizing Systems, Department of Electrical Engineering and Information Technology, Technical University of Darmstadt, 64283 Darmstadt, Germany.
  • Suess B; Centre for Synthetic Biology, Technical University of Darmstadt, 64289 Darmstadt, Germany.
Biosensors (Basel) ; 14(1)2024 Jan 22.
Article en En | MEDLINE | ID: mdl-38275309
ABSTRACT
To combat the growing threat of antibiotic resistance, environmental testing for antibiotic contamination is gaining an increasing role. This study aims to develop an easy-to-use assay for the detection of the fluoroquinolone antibiotic levofloxacin. Levofloxacin is used in human and veterinary medicine and has been detected in wastewater and river water. An RNA aptamer against levofloxacin was selected using RNA Capture-SELEX. The 73 nt long aptamer folds into three stems with a central three-way junction. It binds levofloxacin with a Kd of 6 µM and discriminates the closely related compound ciprofloxacin. Furthermore, the selection process was analyzed using a next-generation sequencing approach to better understand the sequence evolution throughout the selection. The aptamer was used as a bioreceptor for the development of a lateral flow assay. The biosensor exploited the innate characteristic of RNA Capture-SELEX to select aptamers that displace a complementary DNA oligonucleotide upon ligand binding. The lateral flow assay achieved a limit of visual detection of 100 µM. While the sensitivity of this assay constrains its immediate use in environmental testing, the present study can serve as a template for the selection of RNA aptamer-based biosensors.
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Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Aptámeros de Nucleótidos Límite: Humans Idioma: En Revista: Biosensors (Basel) Año: 2024 Tipo del documento: Article País de afiliación: Alemania Pais de publicación: Suiza

Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Aptámeros de Nucleótidos Límite: Humans Idioma: En Revista: Biosensors (Basel) Año: 2024 Tipo del documento: Article País de afiliación: Alemania Pais de publicación: Suiza