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Development of a novel genetically modified bioluminescent-bacteria-based assay for detection of fluoroquinolones in animal-derived foods
- Source :
- Analytical and bioanalytical chemistry. 406(30)
- Publication Year :
- 2014
-
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.
- Subjects :
- Meat
Danofloxacin
Swine
Biosensing Techniques
Biochemistry
Pefloxacin
Analytical Chemistry
Sarafloxacin
Marbofloxacin
Genes, Reporter
Limit of Detection
medicine
Enrofloxacin
Escherichia coli
Animals
Luciferases
Promoter Regions, Genetic
Difloxacin
Chromatography
Chemistry
Escherichia coli Proteins
Anti-Bacterial Agents
Rec A Recombinases
Milk
Luminescent Measurements
Lomefloxacin
Cattle
Orbifloxacin
Genetic Engineering
Chickens
Food Analysis
medicine.drug
Fluoroquinolones
Subjects
Details
- ISSN :
- 16182650
- Volume :
- 406
- Issue :
- 30
- Database :
- OpenAIRE
- Journal :
- Analytical and bioanalytical chemistry
- Accession number :
- edsair.doi.dedup.....4b844fd886c5c0de4f8098c71d177518