Simultaneous and rapid detection of six different mycotoxins using an immunochip
ABSTRACT Mycotoxins are highly toxic contaminants in food, animal feed, and commodities. The study has developed an immunochip for quantifying the concentrations of six mycotoxins: aflatoxin B1, aflatoxin M1, deoxynivalenol, ochratoxin A, T-2 toxin, and zearalenone, which were added to drinking water. The complete antigens (Ags) of the mycotoxins were contact printed and immobilized onto agarose-modified glass slides with 12 physically isolated subarrays, based on the reaction of both diffusion and covalent bond. The optimal concentration of each antigen and antibody (Ab) was obtained using an Ag-Ab immunoassay. Based on the indirect competitive immunoassay for the simultaneous detection of six mycotoxins in one single chip, six standard curves with good logistic correlation (R(2)>0.97) were respectively plotted. The working ranges (0.04-1.69, 0.45-3.90, 20.20-69.23, 35.68-363.18, 0.11-1.81, and 0.08-7.47 ng/mL, respectively) were calculated, as well as the median inhibitory concentrations (0.31±0.04, 1.49±0.21, 34.54±1.30, 134.06±11.75, 0.49±0.05, and 1.54±0.22 ng/mL, respectively), when six mycotoxins were detected simultaneously. Finally, the recovery rates in drinking water generally ranged from 80% to 120% on the same chip, with an intra-assay coefficient of variation lower than 15%. We successfully established an immunochip for simultaneous detection of six mycotoxins within 4h, with advantages of using minimal samples and being visually semiquantitative with our naked eyes. In summary, the method could be developed on one single chip for detecting multiple contaminants in actual samples.
- SourceAvailable from: Zhanhui Wang
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- "Thus, it is required to establish a simple, rapid, low-cost, and sensitive method for routine screening method for monitoring purposes (He et al. 2012). Nowadays, immunochemical assays are the most common available method for routine screening analysis of mycotoxins in a large number of samples (Rosi et al. 2007; Lee 2004; Kolosova et al. 2006; Pei et al. 2009; Guan et al. 2011; He et al. 2012; Wang et al. 2012). The lateral flow strips and ELISA methods are common screening methods. "
ABSTRACT: Aflatoxins are a class of highly toxic chemical contaminants occurring in food. Consumption of aflatoxin-contaminated food can lead to harmful effects on human health. A rapid and reliable analysis of aflatoxins in food is crucial. In this study, we generated a broad–specific monoclonal antibody (MAb) 3 C10 against aflatoxin B1 (AFB1). The MAb 3 C10 binds specifically to AFB1 and AFM1 and has a IC50 of 0.13 μg L−1 for AFB1 and 0.16 μg L−1 for AFM1. Furthermore, the MAb showed high cross-reactivity to AFB2, AFG1, and AFG2. To enable simultaneous AFB1 and AFM1 detection in different food matrices, an indirect competitive enzyme-linked immunosorbent assay (icELISA) based on MAb 3 C10 has been developed and optimized. In addition, the extraction methods of different food matrices (peanut, corn, soybean, wheat flour, rice, soy sauce, vinegar, wine, raw milk, pure milk, skimmed milk, and yogurt) were established. The average recovery ranged from 73 to 121 %, with relative standard deviation values less than 15 %. The limit of detection was 0.52 + 0.36 μg kg−1 (mean + 3SD) for AFB1 in eight agricultural products and 0.031 + 0.015 μg kg−1 (mean + 3SD) for AFM1 in four dairy products. The sensitivity of icELISA was below the limit set by the European Commission for aflatoxin detection in different food matrices and similar to LC–MS/MS method. We demonstrate a rapid, simple, and reliable method for simultaneous screening of AFB1 and AFM1 in different food matrices.Food Analytical Methods 06/2012; 6(3). DOI:10.1007/s12161-012-9484-5 · 1.96 Impact Factor
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ABSTRACT: This review highlights developments in mycotoxin analysis and sampling over a period between mid-2010 and mid- 2011. It covers the major mycotoxins: aflatoxins, Alternaria toxins, ergot alkaloids, fumonisins, ochratoxin, patulin, trichothecenes, and zearalenone. Analytical methods for mycotoxins continue to be developed and published. Despite much interest in immunochemical methods and in the rapid development of LC-MS methodology, more conventional methods, sometimes linked to novel clean-up protocols, have also been the subject of research publications over the above period. Occurrence of mycotoxins falls outside the main focus of this review; however, where relevant to analytical method development, this has been mentioned.World Mycotoxin Journal 02/2012; 5(1):3-30. DOI:10.3920/WMJ2011.1338 · 2.16 Impact Factor
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ABSTRACT: Mycotoxins are highly toxic contaminants in foodstuffs and feedstuffs. The study presents a novel suspension array technology for quantifying four mycotoxins, namely, aflatoxin B1, deoxynivalenol, T-2 toxin, and zearalenone, in corn and peanut. Using 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide hydrochloride, the complete antigens of the mycotoxins became attached to the microspheres with viable activity. The optimal concentrations of each antibody and biotin-rabbit anti-goat IgG were obtained through chessboard titration. The four mycotoxins were detected simultaneously and quantitatively in corn and peanut using indirect competitive immunoassay. Multi-channel standard curves with appropriate logistic correlation (R(2)>0.9819) were respectively plotted. The broad working ranges with three to four orders of magnitude were calculated, and limits of detection at the pg level were found to be better than those obtained using high-performance liquid chromatography. The recovery rates in the actual samples generally ranged from 80.16% to 117.65%, with an intra-assay coefficient of variation lower than 15%, which indicated high accuracy and repeatability. A suspension array method for the simultaneous detection of the four mycotoxins within 4h was successfully developed using minimal samples; the method was proven to have high throughput, flexibility, accuracy and reproducibility. The approach could detect multiple contaminants in actual samples.Biosensors & Bioelectronics 09/2012; 41(1). DOI:10.1016/j.bios.2012.08.057 · 6.41 Impact Factor