Dual-Wavelength Fluorescence Polarization Immunoassay for Simultaneous Detection of Sulfonamides and Antibacterial Synergists in Milk

Abstract

Combinations of sulfonamides (SAs) and antibacterial synergists (ASGs) are frequently used for treating infectious diseases and promoting growth for animals, which cause potential hazards to food safety and human health. To realize the simultaneous detection of SAs and ASGs in food, a homogeneous and high-throughput screening dual-wavelength fluorescence polarization immunoassay (DWFPIA) was developed. In this study, three SAs tracers and three ASGs tracers were synthesized by fluoresceins with different linkers and paired with their corresponding monoclonal antibodies (mAbs), respectively. To achieve a high sensitivity and broad specificity, the combination of tracers SADMPM-HDF with the longest linker paring mAb 10E6 for SAs and tracer HaptenA-DSCA paring mAb 9C9 for ASGs were chosen for the development of DWFPIA, achieving surprising IC50 values for 23 SAs below 100 μg L−1 and 5 ASGs below 50 μg L−1. The accuracy of DWFPIA was applied in real milk samples by typical sulfamethazine (SMZ) and trimethoprim (TMP), with recoveries of 81.7–97.2% and 78.6–103.6%, and coefficient of variations (CVs) below 18.9%, which could be completed within 15 min, including sample pretreatment. We firstly developed a simultaneous screening DWFPIA, covering all of the SAs and ASGs used in clinic and providing a great application potential in food safety analysis.

A Sensitive Fluorescence Polarization Immunoassay for the Rapid Detection of Okadaic Acid in Environmental Waters

“…Currently, many studies have been published on the development of FPIAs using various toxicants, for example, mycotoxins, antibiotics, and pesticides [ 25 , 26 , 27 , 28 ]. The analytical application of this approach for the detection of aquatic toxins is limited to only two papers with the reported FPIAs using hepatotoxins microcystin-LR (MC-LR) and nodularin [ 29 , 30 ].…”

Fluorescence Polarization Assay Based on a New Recognition Motif QepA for the One-Step Detection of Fluoroquinolones in Eggs

“…Conventional heterogeneous immunoassay methods that rely on antibodies and antigens can be time-consuming due to the tedious preparation cycle of antibodies and the multiple operation steps involved in the screening process. Immunoassays and pseudoimmunoassays, which combine new biorecognition motifs with fluorescence polarization (FP), are gaining significant attention in the field of food contamination due to their alignment with practicable application process. − FPA is a homogeneous technology with fewer steps of washing and incubation, where the fluorescent probe, analytes, and the recognition reagent are mixed together. This method is more time-saving than instrument-based analysis methods and traditional ELISA.…”

Generation of a Recombinant scFv against Deoxycholic Acid and Its Conversion to a Quenchbody for One-Step Immunoassay

“…The versatility of fluorescence immunoassays has been widely expanded [10,11]. Among them, reagent-free fluorescence immunoassays offer the advantage of rapid and convenient measurements, which are comparable to traditional ELISA [12][13][14].…”