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Merck

In vitro selection of RNA aptamers that selectively bind danofloxacin.

Biochemical and biophysical research communications (2014-05-06)
Seung Ryul Han, Jaehoon Yu, Seong-Wook Lee
ABSTRACT

Danofloxacin is a synthetic fluoroquinolone with broad spectrum antibacterial activity that is used for the treatment of respiratory diseases in animal husbandry. However, danofloxacin has many adverse reactions and is toxic to humans. Especially, it detrimentally affects muscle, central nerve system, peripheral nerve system, liver, and skin in those who ingest foods in which danofloxacin has accumulated. Prescreening and determination of the level of danofloxacin in foods or food products is necessary for human health. Aptamers are composing of oligonucleotides that specifically interact with target molecules. They are emerging as detection/diagnostic ligands. Here, we used the SELEX in vitro selection technology to identify specific and high-affinity RNA aptamers with 2'-fluoro-2'-deoxyribonucleotide modified pyrimidine nucleotides against danofloxacin. Selected RNA aptamers bound specifically to danofloxacin, but not to tetracycline. Truncation of RNA aptamer up to 36 mer did not comprise specificity and affinity. The truncated RNA aptamer specifically bound to target chemical, allowing the discrimination of danofloxacin from other fluoroquinolones. The isolated specific aptamer could be a potential agent used for the rapid and cost-effective detection and sensing of danofloxacin, replacing instrumental methods including the more expensive and time-consuming methods of high performance liquid chromatography and liquid chromatography/mass spectrometry.

MATERIALS
Product Number
Brand
Product Description

Supelco
Marbofloxacin, VETRANAL®, analytical standard
Sigma-Aldrich
Pyrimidine, ≥98.0%
Sigma-Aldrich
Tetracycline, 98.0-102.0% (HPLC)
Sigma-Aldrich
Tetracycline, 98.0-102.0% (HPLC)
Enrofloxacin for system suitability, European Pharmacopoeia (EP) Reference Standard
Marbofloxacin, European Pharmacopoeia (EP) Reference Standard
Supelco
Enrofloxacin, VETRANAL®, analytical standard
Supelco
Danofloxacin, VETRANAL®, analytical standard
Supelco
Ciprofloxacin, Pharmaceutical Secondary Standard; Certified Reference Material
Sigma-Aldrich
Ciprofloxacin, ≥98% (HPLC)
Sigma-Aldrich
Enrofloxacin, ≥99.0%
Enrofloxacin, European Pharmacopoeia (EP) Reference Standard
Supelco
Ciprofloxacin, VETRANAL®, analytical standard
Ciprofloxacin, European Pharmacopoeia (EP) Reference Standard
USP
Enrofloxacin, United States Pharmacopeia (USP) Reference Standard
Supelco
Enrofloxacin, Pharmaceutical Secondary Standard; Certified Reference Material