title: Novel multiplex fluorescent immunoassays based on quantum dot nanolabels for mycotoxins determination.
abstract: The aim of this manuscript was the development of easy-to-operate quantum dots (QDs)-based immunochemical techniques for simultaneous screening of several mycotoxins in cereals. Two different approaches for multiplex fluorescent immunosorbent assay (FLISA) were employed. In the first approach a multiwell plate in which the different wells express a different mycotoxin (deoxynivalenol, zearalenone, aflatoxin B1, T2-toxin and fumonisin B1) was considered as a multiplex because each sample was pretreated once and then will be distributed over a series of wells within the same plate (single-analyte multiplex, SAM). The entire assay allows the simultaneous determination of all compounds. For the double-analyte multiplex (DAM) two different specific antibodies were co-immobilized in one single well. Zearalenone and aflatoxin B1 were simultaneously determined, provided their conjugates are labeled with QDs which are fluorescent in different parts of the spectrum, by scanning the assay outcome at two different wavelengths. The limits of detection (LOD) for the simultaneous determination of deoxynivalenol, zearalenone, aflatoxin B1, T2-toxin and fumonisin B1 by SAM FLISA were 3.2, 0.6, 0.2, 10 and 0.4 µg kg(-1), respectively, while for the DAM FLISA they were 1.8 and 1 µg kg(-1) for zearalenone and aflatoxin B1, respectively. SAM FLISA principle was also presented in a qualitative on-site format and tested for on-site multiplex determination of four mycotoxins in cereals. The achieved cut-off values of 500, 100, 2 and 100 µg kg(-1) for deoxynivalenol, zearalenone, aflatoxin B1 and T2-toxin respectively. For simplification of multiassay results' evaluation the conjugates with QDs of different colors were used.
authors: Ghent University, Faculty of Pharmaceutical Sciences, Laboratory of Food Analysis, Harelbekestraat 72, 9000 Ghent, Belgium. Electronic address: beloglazovanv@gmail.com.
authors: N V
authors: NV
authors: Beloglazova
authors: Ghent University, Faculty of Pharmaceutical Sciences, Laboratory of Food Analysis, Harelbekestraat 72, 9000 Ghent, Belgium.
authors: E S
authors: ES
authors: Speranskaya
authors: Institute for Agrifood Standards and Testing Technology, Shanghai Academy of Agricultural Sciences, 1018 Jinqi Road, Shanghai 201403, China.
authors: A
authors: A
authors: Wu
authors: College of Veterinary Medicine, National Center for Veterinary Drug Safety Evaluation, China Agricultural University, Yuanmingyuan Western Road 2, 100094 Beijing, China.
authors: Z
authors: Z
authors: Wang
authors: Ghent University, Faculty of Pharmaceutical Sciences, Laboratory of Food Analysis, Harelbekestraat 72, 9000 Ghent, Belgium.
authors: M
authors: M
authors: Sanders
authors: Ghent University, Faculty of Pharmaceutical Sciences, Laboratory of Food Analysis, Harelbekestraat 72, 9000 Ghent, Belgium; Saratov State University, Chemistry Faculty, Department of General and Inorganic Chemistry, Astrakhanskaya 83, 410012 Saratov, Russia.
authors: V V
authors: VV
authors: Goftman
authors: School of Life Science and Biotechnology, Shanghai Jiao Tong University, Dongchuan Road 800, Shanghai 200240, China.
authors: D
authors: D
authors: Zhang
authors: Saratov State University, Chemistry Faculty, Department of General and Inorganic Chemistry, Astrakhanskaya 83, 410012 Saratov, Russia.
authors: I Yu
authors: IY
authors: Goryacheva
authors: Ghent University, Faculty of Pharmaceutical Sciences, Laboratory of Food Analysis, Harelbekestraat 72, 9000 Ghent, Belgium.
authors: S
authors: S
authors: De Saeger
mesh_terms: D055910:Antibodies, Immobilized / Q000737:chemistry; D015374:Biosensing Techniques / Q000191:economics / Q000295:instrumentation*; D002523:Edible Grain / Q000382:microbiology*; D004867:Equipment Design; D007118:Immunoassay / Q000191:economics / Q000295:instrumentation*; D057230:Limit of Detection; D009183:Mycotoxins / Q000032:analysis*; D045663:Quantum Dots / Q000737:chemistry*
chemical_list: D055910:Antibodies, Immobilized; D009183:Mycotoxins
keywords: Immunoassay; Multiassay; Multiplex screening; Mycotoxin; Quantum dots; Rapid tests