Abstract
Enrofloxacin, a veterinary antibiotic that persists in food, poses a risk to human health. Here, a monoclonal antibody against enrofloxacin, 1H12, was prepared based on the hapten ENR-1, and showed excellent sensitivity with a 50% inhibitory concentration (IC50) of 0.03 ng/mL. Using this antibody, 2 lateral-flow immunochromatographic assays were developed for determination of enrofloxacin in egg, milk, honey, and chicken meat samples. The detection ranges (IC20-IC80) were 0.16-0.82 ng/g, 0.24-1.8 ng/g, 0.25-3.6 ng/g, and 0.61-3.9 ng/g by colloidal gold-immunochromatographic sensor (CG-ICS) analysis, and 0.022-0.42 ng/g, 0.054-0.42 ng/g, 0.069-1.4 ng/g, and 0.19-2.2 ng/g by Eu-fluorescence-immunochromatographic sensor (EF-ICS) analysis. The intraassay and interassay recovery rates were 88.9 to 108.5% with coefficients of variation of 1.3 to 7.0% by CG-ICS analysis, and 88.6 to 113.6% with coefficients of variation of 1.3 to 8.1% by EF-ICS analysis. Thus, our newly developed ICS are sensitive and reliable, providing an option for rapid quantitative detection of enrofloxacin in food samples.
Highlights
Enrofloxacin (ENR; Figure 1a) is an important member of the fluoroquinolones and possesses excellent antibacterial effects against gram-positive bacteria, gram-negative bacteria, and mycoplasma by inhibiting DNA-gyrase, which is an essential enzyme in prokaryotes (Zhang et al, 2011; Wang et al, 2014)
The results showed that the visual colorimetric value (vCMV) from colloidal gold-immunochromatographic sensor (CG-immunochromatographic sensor (ICS)) analysis was 0.31 ng/g for egg, milk, and honey samples, and 1.3 ng/g for chicken
We prepared a highly specific and sensitive mAb, 1H12, against ENR based on the hapten ENR-1, and OFL-OVA was selected as the perfect coating antigen with an IC50 of 0.03 ng/mL in ic-ELISA
Summary
Enrofloxacin (ENR; Figure 1a) is an important member of the fluoroquinolones and possesses excellent antibacterial effects against gram-positive bacteria, gram-negative bacteria, and mycoplasma by inhibiting DNA-gyrase, which is an essential enzyme in prokaryotes (Zhang et al, 2011; Wang et al, 2014). Instrumental analysis is currently the most widely used method for monitoring ENR residues due to its high sensitivity and accuracy, with methods including HPLC (Aufartova et al, 2017; Huang et al, 2018), liquid chromatography (LC)-MS (Lolo et al, 2005; Panzenhagen et al, 2016), and surface plasmon resonance (Fernandez et al, 2010). These methods require expensive equipment, professional skill, and complicated sample pretreatment, but are low-throughput and time-consuming techniques (Meng and Xi, 2011). It is difficult to meet the need for on-site screening of large numbers of samples using these approaches
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