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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    • "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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