title: Paper-based colorimetric biosensor for antibiotics inhibiting bacterial protein synthesis.
abstract: Due to the presence of antibiotics in environmental water and their potential influence on the occurrence of antibiotic-resistant bacteria, development of a detection method suitable for the screening of environmental water for antibiotics is required. In this study, we developed a simple colorimetric paper-based biosensor based on a novel principle for the detection of antibiotics inhibiting bacterial protein synthesis, including aminoglycosides, tetracycline, chloramphenicol, and macrolides. This biosensor is based on the detection of a color change induced by β-galactosidase, which is synthesized on freeze-dried paper discs containing an in vitro transcription/translation system. When a water sample without antibiotics is applied to the paper discs, β-galactosidase can be synthesized, and it hydrolyzes a colorimetric substrate, resulting in a color change from yellow to purple. By contrast, in the presence of antibiotics, the color change can be hampered due to an inhibition of β-galactosidase synthesis. We investigated the effect of the incubation temperature and pH of water samples and confirmed that the paper discs showed the color change to purple in the ranges of 15-37°C and pH 6-10. We observed concentration-dependent color variations of the paper discs by the naked eye and further estimated detection limits to be 0.5, 2.1, 0.8, and 6.1 μg/mL for paromomycin, tetracycline, chloramphenicol, and erythromycin, respectively, using digitized pictures. The paper-based biosensor proved to detect 0.5 μg/mL paromomycin, spiked in real environmental water samples, by the naked eye.
authors: Graduate School of Pharmaceutical Sciences, Osaka University, 1-6 Yamadaoka, Suita, Osaka 565-0871, Japan; College of Aquaculture and Fisheries, Can Tho University, Campus II, 3/2 Street, Can Tho City, Viet Nam.
authors: Tran Thi My
authors: TT
authors: Duyen
authors: Graduate School of Pharmaceutical Sciences, Osaka University, 1-6 Yamadaoka, Suita, Osaka 565-0871, Japan. Electronic address: hmhide@phs.osaka-u.ac.jp.
authors: Hideyuki
authors: H
authors: Matsuura
authors: Graduate School of Pharmaceutical Sciences, Osaka University, 1-6 Yamadaoka, Suita, Osaka 565-0871, Japan.
authors: Kazuki
authors: K
authors: Ujiie
authors: Graduate School of Pharmaceutical Sciences, Osaka University, 1-6 Yamadaoka, Suita, Osaka 565-0871, Japan.
authors: Misa
authors: M
authors: Muraoka
authors: Graduate School of Pharmaceutical Sciences, Osaka University, 1-6 Yamadaoka, Suita, Osaka 565-0871, Japan.
authors: Kazuo
authors: K
authors: Harada
authors: Graduate School of Pharmaceutical Sciences, Osaka University, 1-6 Yamadaoka, Suita, Osaka 565-0871, Japan.
authors: Kazumasa
authors: K
authors: Hirata
mesh_terms: D000900:Anti-Bacterial Agents / Q000032:analysis* / Q000494:pharmacology*; D001419:Bacteria / Q000187:drug effects* / Q000378:metabolism*; D001426:Bacterial Proteins / Q000096:biosynthesis*; D015374:Biosensing Techniques / Q000379:methods*; D003124:Colorimetry / Q000379:methods*
chemical_list: D000900:Anti-Bacterial Agents; D001426:Bacterial Proteins
keywords: Cell-free protein synthesis; Chloramphenicol; Chlorophenol red-β-d-galactopyranoside; Environmental water; Erythromycin; Paper-based biosensor; Paromomycin; Resistant bacteria; Tetracycline; β-Galactosidase