Development of an indirect competitive ELISA for simultaneous detection of enrofloxacin and ciprofloxacin

Abstract

Modified 1-ethyl-3-(3-dimethylaminopropy) carbodiimide (EDC) method was employed to synthesize the artificial antigen of enrofloxacin (ENR), and New Zealand rabbits were used to produce anti-ENR polyclonal antibody (pAb). Based on the checkerboard titration, an indirect competitive enzyme-linked immunosorbent assay (ELISA) standard curve was established. This assay was sensitive and had a linear range from 0.6 to 148.0 μg/kg (R(2) = 0.9567), with the half maximal inhibitory concentration (IC(50)) and limit of detection (LOD) values of 9.4 μg/kg and 0.2 μg/kg, respectively. Of all the competitive analogues, the produced pAb exhibited a high cross-reactivity to ciprofloxacin (CIP) (87%), the main metabolite of ENR in tissues. After optimization, the matrix effects can be ignored using a 10-fold dilution in beef and 20-fold dilution in pork. The overall recoveries and coefficients of variation (CVs) were in the ranges of 86%-109% and 6.8%-13.1%, respectively. It can be concluded that the established ELISA method is suitable for simultaneous detection of ENR and CIP in animal tissues.

A Simple DNA-based Electrochemical Biosensor for Highly Sensitive Detection of Ciprofloxacin Using Disposable Graphene

“…3 Conventional analytical procedures of drug residues have heavily depended on traditional methods, such as high-performance liquid chromatography and liquid chromatography-mass spectrometry, which are laborious and time-consuming and unsuitable for screening large number of samples. 4,5 Thus, there is an urgent need for a highly sensitive and rapid screening ciprofloxacin device to measure environmental contamination. Lately, recent developments in DNA-based biosensors have focused on the interaction of drugs and DNA, providing a convenient method for detecting drugs, possible miniaturization as well as portability.…”

Rapid screening and quantification of multi-class antibiotic pollutants in water using a planar waveguide immunosensor

“…Analytical techniques used to quantify antibiotic concentrations in food and environmental samples include highperformance liquid chromatography (HPLC), 9,10 HPLC coupled with mass spectrometry (HPLC/MS) 11 and enzyme-linked immunosorbent assays (ELISAs). 12,13 However, these methods are time-consuming and expensive, and specialized laboratory instrument is required for the detection process. 14…”

Development of an ic-ELISA and immunochromatographic strip for detection of sparfloxacin in honey

“…Many analytical methods have been used to detect fluoroquinolones, including high-performance liquid chromatography (HPLC), liquid chromatography-tandem mass spectrometry, fluorescence analyses, spectrophotometric analyses, and capillary electrophoresis (Alcaráz, Siano, Culzoni, Muñoz de la Peña, & Goicoechea, 2014;Gul, Sultana, Saeed Arayne, Shamim, & Akhtar, 2012;He & Blaney, 2015;Li et al, 2009;Pinacho, Sánchez-Baeza, Pividori, & Marco, 2014), which are all complex, time-consuming, and expensive. Compared with these methods, an immunoassay is a sensitive, low-cost, and rapid technique that has already been reported to detect fluoroquinolones (Chen & Jiang, 2013;Jiang et al, 2011;Liu et al, 2014;Peng, Liu, Kuang, Cui, & Xu, 2017;Tian, Zhang, Sun, Qian, & Ji, 2013;Tochi, Khaemba, et al, 2016;Wang, Zhang, Ni, Zhang, & Shen, 2014;Zhang et al, 2011). Although the enzymelinked immunosorbent assay (ELISA) with a half maximal inhibitory concentration (IC 50 ) of 31 ng/mL was established (Deng et al, 2013), it does not meet the requirements for field applications.…”