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Development of a novel genetically modified bioluminescent-bacteria-based assay for detection of fluoroquinolones in animal-derived foods

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Abstract

Fluoroquinolones (FQNs) are broad-spectrum antibacterial agents widely used in animal husbandry and aquaculture. The residues and antimicrobial resistance of such antibiotics are a major public health concern. To realize multianalyte detection of FQN residues, a genetically modified bacterium, Escherichia coli pK12 harboring plasmid pRecAlux3, was constructed in this study to develop a bioluminescent-bacteria-based assay for the detection of FQNs in animal-derived foods. This assay was based on the principle of induction of an SOS response by FQNs via inducing the recA-promoter-fused luciferase reporter gene existing on the plasmid pRecAlux3. E. coli pK12 was able to recognize 11 FQNs: difloxacin, enrofloxacin, ciprofloxacin, sarafloxacin, norfloxacin, danofloxacin, ofloxacin, pefloxacin, lomefloxacin, marbofloxacin, and orbifloxacin. This method could be applied to 11 edible tissues, including milk, fish muscle, and the muscles, livers, and kidneys of cattle, chickens, and pigs, with a very simple and rapid sample extraction procedure using only phosphate-buffered saline. The limits of detection of the FQNs were between 12.5 and 100 μg kg−1, all of which were lower than the maximum residue limits. Most of the recoveries of the FQNs were in the range from 60 to 120 %, and the interassay coefficients of variation were less than 30 %. This method, confirmed by high-performance liquid chromatography, is reliable and can be used as both a screening test and a semiquantitative assay, when the identity of a single type of FQN is known.

The principle of the genetically modified bioluminescent bacteria based assay for detection offluoroquinolones

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Abbreviations

BLBA:

Bioluminescent-bacteria-based assay

CV:

Coefficient of variation

FQN:

Fluoroquinolone

GFP:

Green fluorescent protein

HPLC:

High-performance liquid chromatography

IC:

Induction coefficient

LB:

Luria–Bertani

LOD:

Limit of detection

MRL:

Maximum residue level

PBS:

Phosphate-buffered saline

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Acknowledgments

This work was supported by the Program of International S & T Cooperation Funded Project (program no. 2011DFA32140), the National Natural Science Foundation of China (grant nos. 31272614 and 31302140), and the Fundamental Research Funds for the Central Universities (program nos. 2013QC002 and 2011PY078).

Conflict of interest

The authors declare that they have no conflict of interest.

Author information

Authors and Affiliations

  1. MOA Laboratory for Risk Assessment of Quality and Safety of Livestock and Poultry Products, Huazhong Agricultural University, Wuhan, 430070, China

    Guyue Cheng, Xu Wang, Haihong Hao & Zonghui Yuan

  2. National Reference Laboratory of Veterinary Drug Residues (HZAU) and MOA Key Laboratory for Detection of Veterinary Drug Residues, Huazhong Agricultural University, Wuhan, 430070, China

    Xiaobing Dong, Yulian Wang, Dapeng Peng, Shuyu Xie, Wei Qu, Zhenli Liu & Zonghui Yuan

Authors
  1. Guyue Cheng
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  2. Xiaobing Dong
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  3. Yulian Wang
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  4. Dapeng Peng
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  5. Xu Wang
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  6. Haihong Hao
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  7. Shuyu Xie
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  8. Wei Qu
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  9. Zhenli Liu
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  10. Zonghui Yuan
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Corresponding author

Correspondence to Zonghui Yuan.

Additional information

Guyue Cheng and Xiaobing Dong contributed equally to this work.

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Cheng, G., Dong, X., Wang, Y. et al. Development of a novel genetically modified bioluminescent-bacteria-based assay for detection of fluoroquinolones in animal-derived foods. Anal Bioanal Chem 406, 7899–7910 (2014). https://doi.org/10.1007/s00216-014-8228-3

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  • DOI: https://doi.org/10.1007/s00216-014-8228-3

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