Microchimica Acta

, 186:150 | Cite as

Enzyme-free fluorometric assay for chloramphenicol based on double stirring bar-assisted dual signal amplification

  • Feng Hong
  • Xiaoting Lin
  • Yongxiang WuEmail author
  • Youren Dong
  • Yuting Cao
  • Futao Hu
  • Ning GanEmail author
Original Paper


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).

Graphical abstract

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.


Catalyzed hairpin assembly Thioflavin T Target recycling Food safety Antibiotics detection 



This work was supported by Natural Science Foundation of Zhejiang (LY19B050001,Y18B070008), Zhejiang Province Welfare Technology Applied Research Project (LGN18H300001), the Natural Science and Huiming Foundation of Ningbo (2017C50035), Natural Science Foundation of Zhejiang (LY17C200007), Zhejiang Province Welfare Technology Applied Research Project (2017C37023), Open Fund of Key Laboratory of Marine New Materials and Applied Technology in Chinese Academy of Sciences(2018K08) and K. C. Wong Magna Fund in Ningbo University.

Compliance with ethical standards

The author(s) declare that they have no competing interests.

Supplementary material

604_2018_3148_MOESM1_ESM.doc (622 kb)
ESM 1 (DOC 621 kb)


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Copyright information

© Springer-Verlag GmbH Austria, part of Springer Nature 2019

Authors and Affiliations

  1. 1.Faculty of Material Science and Chemical EngineeringNingbo UniversityNingboChina
  2. 2.Faculty of MarineNingbo UniversityNingboChina

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