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- Title
Enzyme-free fluorometric assay for chloramphenicol based on double stirring bar-assisted dual signal amplification.
- Authors
Hong, Feng; Lin, Xiaoting; Wu, Yongxiang; Dong, Youren; Cao, Yuting; Hu, Futao; Gan, Ning
- Abstract
An enzyme-free fluorometric assay is described that accomplishes dual signal amplification by making use of a two stirring bars. Two Y-shaped DNA probes were designed and placed on the bars. When the target (with chloramphenicol as model analyte) is added, it triggers target recycling and simultaneously catalyzes hairpin assembly (CHA). A large fraction of DNA primers is released by the analyte from the bar to the supernatant and open hairpins with G-quadruplex DNA sequence. The G-quadruplex can specifically bind thioflavin T (ThT) to emit fluorescence (with excitation/emission maxima at 445 and 485 nm) for quantification of chloramphenicol. An enzyme is not needed. ThT is added to the system as a fluorescent DNA probe. All this strongly reduces the cost for sensor construction and usage. The dual signal amplification steps occur simultaneously which reduces the detection time. The assay was successfully employed to the determination of CAP in spiked milk and fish samples within 60 min and with a 16 pM limit of detection (at S/N = 3).Schematic representation of a new method for the detection of chloramphenicol by using two stirring bars. It is based on target recycling and catalyzed hairpin assembly amplification. CAP: chloramphenicol, ThT: thioflavin T, CHA: catalyzed hairpin assembly.
- Subjects
DNA primers; CHLORAMPHENICOL; DNA probes; NUCLEOTIDE sequence; FLUORESCENT probes; DETECTION limit
- Publication
Microchimica Acta, 2019, Vol 186, Issue 3, p1
- ISSN
0026-3672
- Publication type
Article
- DOI
10.1007/s00604-018-3148-0