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Abstract
A new highly specific and sensitive monoclonal antibody (MAb) to T-2 toxin (T-2) was produced, providing an IC50 value of 1.02 ng/mL and negligible cross-reactivity (CR) to other related mycotoxins. Based on the new MAb, a lateral-flow immunochromatographic assay (LFIA) using colloidal gold (CG) and fluorescent microspheres (FMs) as labels was proposed for T-2. Under the optimized conditions, in rapid qualitative assay, the cut-off values of the CG-LFIA were 400 μg/kg in rice and 50 μg/L in fresh milk, and the cut-off values of the FMs-LFIA were 100 μg/kg in both rice and chicken feed. For the quantitative assay with the FMs-LFIA, the limit of detection (LOD) were 0.23 μg/kg and 0.41 μg/kg in rice and chicken feed, respectively, and the average recoveries ranged from 80.2% to 100.8% with the coefficient of variation (CV) below 10.8%. In addition, we found that the CG-LFIA could tolerate the matrix effect of fresh milk better than the FMs-LFIA, while the FMs-LFIA could tolerate the matrix effect of chicken feed better than CG-LFIA under the same experimental conditions. These results provide a certain reference for the selection of appropriate labels to establish a rapid LFIA in various biological samples.
Highlights
T-2 toxin (T-2), a type A trichothecene, is mainly produced by Fusarium species [1].It is a ubiquitous contaminant of cereals and processed foods, occurring mainly in cold climate regions or during wet storage conditions [2,3,4]
T-2 toxin, HT-2 toxin, deoxynivalenol (DON), neosolaniol (NEO), nivalenol (NIV), T-2-triol, T-2-tetraol, keyhole limpet hemocyanin (KLH), bovine serum albumin (BSA), ovalbumin (OVA), Freund’s incomplete adjuvant (FIA), Freund’s complete adjuvant (FCA), PEG1500, hypoxanthine aminopterin thymidine (HAT), and a mouse monoclonal antibody (MAb) isotyping kit were obtained from Sigma-Aldrich
T-2-free fresh milk samples were supplied by the National Reference Laboratory for Veterinary Drug Residues (Beijing, China) and those samples were directly used for analysis by the GC-lateral-flow immunochromatographic assay (LFIA) and fluorescent microspheres (FMs)-LFIA methods without further extraction steps
Summary
T-2 toxin (T-2), a type A trichothecene, is mainly produced by Fusarium species [1]. It is a ubiquitous contaminant of cereals and processed foods, occurring mainly in cold climate regions or during wet storage conditions [2,3,4]. Several analytical methods for detecting T-2 have been reported, including high-performance liquid chromatography (HPLC) with a fluorescence detector and liquid chromatography-tandem mass spectrometry (LC-MS/MS). Are unsuitable for high-throughput metabolites were toxic [10,11,12] Once exposed to those it, no methods effective solution is available to avoid the screening of[13], large of samples because they are timemust consuming andestablished. ELISA method require labor-intensive operations, including incubation, spectrometry Those methods are unsuitable for high-throughput washing and enzymatic reactions [19]. MAb high specificity and sensitivity produced for matrix tolerance in different biological samples has been conducted. Anti-T-2 MAb specificity and sensitivity was produced and LFIAof the proposed assay, thean tolerance of thewith twohigh labels for different matrices was estimated. To fully determine the feasibility of the proposed assay, the tolerance of the two labels for different matrices was estimated
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