Simultaneous and rapid detection of six different mycotoxins using an immunochip

College of Food Science and Technology, Huazhong Agricultural University, Wuhan, PR China.
Biosensors & Bioelectronics (Impact Factor: 6.41). 01/2012; 34(1):44-50. DOI: 10.1016/j.bios.2011.12.057
Source: PubMed


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.

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    • "The imaging surface plasmon resonance (iSPR) platform (Raz, Bremer, Haasnoot, & Norde, 2009) and the wavelength-interrogated optical system (WIOS) (Adrian et al., 2009) have been reported for multiplex detection of antibiotic residues, which is known as the optical waveguide light-mode spectroscopy (OWLS) (Adányi et al., 2007) suspension array (Ying et al., 2013). In addition, immune chip (Ying et al., 2012) has also been developed for simultaneous detection of mycotoxins. These methods however are limited in detecting multiplex compounds in a single class, while some need more than four hours for the analysis, which are not suitable for an onsite , low cost, and real time detection, as opposed to our present study. "
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    ABSTRACT: Mycotoxins and industrial chemicals, such as aflatoxin M1 and melamine, now commonly exist in milk and cause potential health risks. This study presents an indirect competitive immunoassay through multiplex planar waveguide fluorescence immunosensor (MPWFI) for rapid, sensitive, and simultaneous detection and quantification of aflatoxin M1 and melamine by applying the principle of immunoreaction and total internal reflect fluorescent. Double-channel standard curves with appropriate logistic correlation (R2 > 0.99) were plotted, respectively. The working ranges (0.073-0.400 ng/mL and 26.38-270.00 ng/mL, respectively) were calculated, as well as the limit of detection (0.045 and 13.37 ng/mL, respectively), when two analytes were simultaneously detected. Both results satisfied the requirements for the maximum amount set by the WHO, which illustrated that the current method was better than some other standard methods. The recovery rates in the actual samples ranged from 85% to 103%, with relative standard deviations between 1.3% and 6.5%, which indicated high accuracy and repeatability.
    Full-text · Article · Aug 2015 · Food Chemistry
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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. "
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    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.
    Full-text · Article · Jun 2012 · Food Analytical Methods
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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.
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