Development of a competitive immunochromatographic assay for the sensitive detection of amantadine in chicken muscle.

Amantadine (AMD) is a prohibitive veterinary medicine in the entire world. In this study, a sensitive colloidal gold immunochromatographic assay (CGICA) was established for the rapid semi-quantitative detection of AMD in chicken muscle. Under optimal conditions, the detection results were obtained in 12min with a limit of detection for 1.80ng/mL. CGICA presented a good linear range from 2.5ng/mL to 25ng/mL, with only 11.5% cross-reactivity with rimantadine. The recovery rates for the fortified samples were ranged from 81% to 120%. The coefficient of variation of the intra-assay and inter-assay was less than 15%. The accuracy of CGICA was confirmed by systematically comparing the result of the proposed method with enzyme-linked immunosorbent assay and ultra-high-performance liquid chromatography-tandem mass spectrometry. Given the advantages of its simplicity, convenience, and speediness, the proposed CGICA is suitable for the on-site rapid detection of AMD in chicken muscle.

Comparison of immunochromatographic assays based on fluorescent microsphere and quantum-dot submicrobead for quantitative detection of aflatoxin M1 in milk.

The performance of fluorescent microsphere immunochromatographic assay (FM-ICA) and quantum-dot submicrobead immunochromatographic assay (QB-ICA) was systematically and comprehensively compared in quantitative detection of aflatoxin M1 in milk. Under optimum conditions, the advantages of FM-ICA include lower limit of detection of 42.3 pg/mL with better accuracy, precision, reliability, and practicability. The advantages of QB-ICA include shorter detection time and lower monoclonal antibody consumption. The 2 ICA were consistent with liquid chromatography-tandem mass spectrometry. This study serves as a reference for selecting appropriate fluorescent labels for the immunochromatographic assay of aflatoxin M1.

Advantages of time-resolved fluorescent nanobeads compared with fluorescent submicrospheres, quantum dots, and colloidal gold as label in lateral flow assays for detection of ractopamine.

Label selection is a critical factor for improving the sensitivity of lateral flow assay. Time-resolved fluorescent nanobeads, fluorescent submicrospheres, quantum dots, and colloidal gold-based lateral flow assay (TRFN-LFA, FM-LFA, QD-LFA, and CG-LFA) were first systematically compared for the quantitative detection of ractopamine in swine urine based on competitive format. The limits of detection (LOD) of TRFN-LFA, FM-LFA, QD-LFA, and CG-LFA were 7.2, 14.7, 23.6, and 40.1pg/mL in swine urine samples, respectively. The sensitivity of TRFN-LFA was highest. In the quantitative determination of ractopamine (RAC) in swine urine samples, TRFN-LFA exhibited a wide linear range of 5pg/mL to 2500pg/mL with a reliable coefficient of correlation (R2=0.9803). Relatively narrow linear ranges of 10-500pg/mL (FM-LFA) and 25-2500pg/mL (QD-LFA and CG-LFA) were acquired. Approximately 0.005µg of anti-RAC poly antibody (pAb) was used in each TRFN-LFA test strip, whereas 0.02, 0.054, and 0.15µg of pAb were used in each of the FM-LFA, QD-LFA, and CG-LFA test strips, respectively. In addition, TRFN-LFA required the least RAC-BSA antigens and exhibited the shortest detection time compared with the other lateral flow assays. Analysis of the RAC in swine urine samples showed that the result of TRFN-LFA was consistent with that of liquid chromatography-tandem mass spectrometry (LC-MS/MS) and a commercial enzyme-linked immunosorbent assay (ELISA) kit.

Sulfonated polystyrene magnetic nanobeads coupled with immunochromatographic strip for clenbuterol determination in pork muscle.

A magnetic solid-phase extraction method (MSPE) was developed to pre-concentrate and cleanup clenbuterol (CLE) from pork muscle. Novel sulfonated polystyrene magnetic nanobeads (spMNBs) were synthesized via a one-pot emulsion copolymerization method by using divinylbenzene, styrene, and sodium styrene sulfonate in the presence of oleic acid-modified and 10-undecylenic acid-modified magnetic ferrofluid. The resulting spMNBs exhibited high adsorption efficiency for CLE and for 10 other common beta-adrenergic agonists, namely, brombuterol, ractopamine, tulobuterol, bambuterol, cimbuterol, mabuterol, clorprenaline, penbutolol, salbutamol, and cimaterol. The adsorption behavior of the spMNBs for CLE was described by the Langmuir equation with a maximum adsorption capacity of 0.41 mg/g. Under the optimized parameters, the extraction of CLE from 0.5 g of pork muscle required 25mg of the spMNBs at a shortened adsorption time (0.5 min). The proposed MSPE was coupled with colloidal gold nanoparticle-based immunochromatographic assay (MSPE-AuNPIA) for the quantitative detection of CLE residue in pork muscle. The limit of detection and limit of quantification for the pork muscle were 0.10 and 0.24 ng/g, respectively. The intra-day and inter-day assay recoveries at three CLE spiked concentrations ranged from 92.5% to 98.1%, with relative standard deviations ranging from 3.2% to 13.0%. The results of MSPE-AuNPIA were confirmed by liquid chromatography-tandem mass spectrometry (LC-MS/MS). The CLE values obtained with MSPE-AuNPIA agreed with those obtained with LC-MS/MS.