Multiplex Flow Cytometric Immunoassays for High-Throughput Screening of Multiple Mycotoxin Residues in Milk
- 112 Downloads
Microsphere-based immunoassays involving flow cytometry have recently gained popularity for use in protein detection and infectious disease diagnosis due to their simple assay format and capacity for multiplexed analysis. In this work, four mycotoxins, including aflatoxin M1 (AFM1), ochratoxin A (OTA), deoxynivalenol (DON), and fumonisin B1 (FB1), were selected as model analytes. A quadplex flow cytometric immunoassay (FCIA) was developed for detecting multiple mycotoxin residues in milk. The optimized quadplex FCIA exhibited satisfactory compatibility with the multiple mycotoxin residue analysis, with limits of detection (LODs) of 0.045 μg L−1 for AFM1, 0.94 μg L−1 for OTA, 7.48 μg L−1 for DON, and 2.45 μg L−1 for FB1. The recoveries of the target mycotoxins from spiked milk were 76–95%, with a relative standard deviation of less than 13.4%. Compared to traditional ELISA formats, the FCIA provided lower detection limits for multiple mycotoxin detection.
KeywordsMycotoxin Multi-analyte residue Flow cytometric immunoassay Milk Food safety
This work was supported by the National Natural Science Foundation of China (No. 31602103); the Science and Technology Planning Project of Guangdong Province, China (Nos. 2016A020210055 and 2017B020207009); the Shenzhen Basic Research Project (Nos. JCYJ20160307114724751 and JCYJ20170817095240632); and the Shenzhen Sanming Project of Medicine and Health (No. SZSM201611068).
Compliance with Ethical Standards
Conflict of Interest
Jianwen Qu declares that he has no conflict of interest. Huijuan Xie declares that he has no conflict of interest. Shuying Zhang declares that he has no conflict of interest. Pengjie Luo declares that he has no conflict of interest. Ping Guo declares that he has no conflict of interest. Yuebin Ke declares that he has no conflict of interest. Junyu Zhuang declares that he has no conflict of interest. Fangmei Zhou declares that he has no conflict of interest. Wenxiao Jiang declares that he has no conflict of interest.
This article does not contain any studies with human participants or animals performed by any of the authors.
- Aguilera-Luiz MM, Plaza-Bolaños P, Romero-González R, Martínez Vidal JL, Frenich AG (2011) Comparison of the efficiency of different extraction methods for the simultaneous determination of mycotoxins and pesticides in milk samples by ultra high-performance liquid chromatography-tandem mass spectrometry. Anal Bioanal Chem 399:2863–2875. https://doi.org/10.1007/s00216-011-4670-7 CrossRefGoogle Scholar
- Han Z, Ren Y, Zhu J, Cai Z, Chen Y, Luan L, Wu Y (2012) Multianalysis of 35 mycotoxins in traditional Chinese medicines by ultra-high-performance liquid chromatography–tandem mass spectrometry coupled with accelerated solvent extraction. J Agric Food Chem 60:8233–8247. https://doi.org/10.1021/jf301928r CrossRefGoogle Scholar
- He J, Wu N, Luo P, Guo P, Qu J, Zhang S, Zou X, Wu F, Xie H, Wang C, Jiang W (2017) Development of a heterologous enzyme-linked immunosorbent assay for the detection of clindamycin and lincomycin residues in edible animal tissues. Meat Sci 125:137–142. https://doi.org/10.1016/j.meatsci.2016.11.024 CrossRefGoogle Scholar
- Jiang W, Beier RC, Wang Z, Wu Y, Shen J (2013a) Simultaneous screening analysis of 3-methyl-quinoxaline-2-carboxylic acid and quinoxaline-2-carboxylic acid residues in edible animal tissues by a competitive indirect immunoassay. J Agric Food Chem 61:10018–10025. https://doi.org/10.1021/jf4037497 CrossRefGoogle Scholar
- Jiang W, Zhang H, Li X, Liu X, Zhang S, Shi W, Shen J, Wang Z (2013d) Monoclonal antibody production and the development of an indirect competitive enzyme-linked immunosorbent assay for screening spiramycin in milk. J Agric Food Chem 61:10925–10931. https://doi.org/10.1021/jf404027b CrossRefGoogle Scholar
- Jiang W, Beloglazova NV, Wang Z, Jiang H, Wen K, Saeger S, Luo P, Wu Y, Shen J (2015) Development of a multiplex flow-through immunoaffinity chromatography test for the on-site screening of 14 sulfonamide and 13 quinolone residues in milk. Biosens Bioelectron 66:124–128. https://doi.org/10.1016/j.bios.2014.11.004 CrossRefGoogle Scholar
- Liu N, Su P, Gao Z, Zhu M, Yang Z, Pan X, Fang Y, Chao F (2009) Simultaneous detection for three kinds of veterinary drugs: chloramphenicol, clenbuterol and 17-beta-estradiol by high-throughput suspension array technology. Anal Chim Acta 632:128–134. https://doi.org/10.1016/j.aca.2008.10.061 CrossRefGoogle Scholar
- Meneely J, Ricci F, Vesco S, Abouzied M, Sulyok M, Krska R, Elliott C (2011) A comparative study of qualitative immunochemical screening assays for the combined measurement of T-2/HT-2 in cereals and cereal-based products. World Mycotoxin J 4:385–394. https://doi.org/10.3920/WMJ2011.1313 CrossRefGoogle Scholar
- Meng H, Wang Z, De Saeger S, Wang Y, Wen K, Zhang S, Shen J (2014) Determination of ochratoxin a in cereals and feeds by ultra-performance liquid chromatography coupled to tandem mass spectrometry with immunoaffinity column clean-up. Food Anal Method 7:854–864. https://doi.org/10.1007/s12161-013-9692-7 CrossRefGoogle Scholar
- Nardiello D, Lo Magro S, Iammarino M, Palermo C, Muscarella M, Centonze D (2014) Recent advances in the post-column derivatization for the determination of mycotoxins in food products and feed materials by liquid chromatography and fluorescence detection. Curr Anal Chem 10:355–365CrossRefGoogle Scholar
- Zhao Z, Liu N, Yang L, Deng Y, Wang J, Song S, Lin S, Wu A, Zhou Z, Hou J (2015) Multi-mycotoxin analysis of animal feed and animal-derived food using LC–MS/MS system with timed and highly selective reaction monitoring. Anal Bioanal Chem 407:7359–7368. https://doi.org/10.1007/s00216-015-8898-5 CrossRefGoogle Scholar