Ultrasensitive indirect competitive ELISA and strip sensor for detection of furazolidone metabolite in animal tissues

Abstract

Nitrofurans are a class of drugs, which are typically used as antibiotics or antimicrobials illegally in aquiculture. In this paper, indirect competitive enzyme-linked immunosorbent assay and strip sensor were developed for detection of furazolidone metabolite, 3-amino-2-oxazolidinone (AOZ). These two methods could detect AOZ with high sensitivity and specificity. The immunosorbent assay with the half-inhibitory concentration of 0.074 ng/mL has a good recovery from 79.9% to 119.8% for detection of AOZ in pork, chicken, fish, shrimp, pork liver and chicken liver. Based on the developed strip sensor, the detection limit was 0.05 ng/g by the naked eyes and the cut-off value was 0.3 ng/g. The cross-reactivity with other nitrofurans, metabolites and some other compounds was all less than 0.1%. The sample recoveries ranged from 97.3% to 115.5%. Both these methods could be used for on-site detection of furazolidone metabolite at trace level in animal tissues. ARTICLE HISTORY

The Growing Interest in Development of Innovative Optical Aptasensors for the Detection of Antimicrobial Residues in Food Products

“…However, these methods often lack sensitivity for some antimicrobials. Immunological methods like ELISA tests [8,9] and dipsticks [10,11] are implemented for more specific detection of one or several classes of antimicrobial residues or for banned substances. Their sensitivity is sometimes limited, especially since the RPAs are lower [3].…”

Manganese Molybdenum Oxide Micro Rods Adorned Porous Carbon Hybrid Electrocatalyst for Electrochemical Determination of Furazolidone in Environmental Fluids

“…It can be used to treat ulcers and gastrointestinal diseases which are triggered by bacteria and protozoa. However, some research has suggested that furazolidone has pathogen infection in poultry, pigs, rabbits, fish, and human health [1,2]. The economically farmed animals' death is prevented by furazolidone and it is illegally added to animal food in most of the countries [3,4].…”

Plate-Adherent Nanosubstrate for Improved ELISA of Small Molecules: A Proof of Concept Study

“…Enzyme-linked immunosorbent assay (ELISA) is the workhorse of bioanalytical chemistry and serves as the gold standard for monitoring certain diagnostic and prognostic biomarkers, including peptides, proteins, antibodies, hormones, and small molecule metabolites. − The simplicity and versatility of the ELISA technique is attributed to the ability of the capture antigen/antibody to be passively adsorbed onto a polystyrene (PS) well plate, ,, thereby facilitating easy separation of specific and nonspecific binding of assay reagents by periodic washing steps. However, a vigorous washing regimen has often been reported to cause partial desorption of passively adsorbed antigen/antibody from the wells, thereby affecting the sensitivity, limit-of-detection, and reproducibility of the assay. , The propensity of desorption is more pronounced in competitive ELISA, especially when a small molecule is adsorbed onto the well plate.…”

“…Our spiking experiments in artificial urine demonstrated that the improved ELISA technique with Gel–BSA–OHG substrate can specifically detect 8-OHdG with high sensitivity bereft of any interference from the urine matrix. Despite advancements in spectroscopic, chromatographic, and PCR-based techniques for the detection of small molecules, ELISA has been gaining importance owing to its simplicity, low cost, automation, and high throughput. ,, Moreover, the sensitivity of ELISA was being further enhanced by integrating NP as enzyme mimics, enzyme/antibody carriers, and signal transducers . To the best of our knowledge, this is the first study to use NP as a coating substrate for the efficient immobilization of small molecules on PS well plates of ELISA.…”