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Toxins 2018, 10(10), 415; https://doi.org/10.3390/toxins10100415

Antibody Microarray Immunoassay for Simultaneous Quantification of Multiple Mycotoxins in Corn Samples

1
China-Australian Joint Laboratory for Animal Health Big Data Analytics, Zhejiang Provincial Engineering Laboratory for Animal Health Inspection and Internet Technology, College of Animal Science and Technology, Zhejiang A&F University, Lin’an 311300, Zhejiang, China
2
Zhejiang University Institute of Preventive Veterinary Medicine and Zhejiang Provincial Key Laboratory of Preventive Veterinary Medicine, 388 Yuhangtang Road, Hangzhou 310058, Zhejiang, China
3
Technic Center of Zhejiang Entry-Exit Inspection and Quarantine Bureau, 126 Fuchun Road, Hangzhou 310012, Zhejiang, China
*
Author to whom correspondence should be addressed.
Received: 19 September 2018 / Revised: 9 October 2018 / Accepted: 10 October 2018 / Published: 15 October 2018
(This article belongs to the Collection Biorecognition Assays for Mycotoxins)
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Abstract

We developed and tested a prototype of an antibody microarray immunoassay for simultaneous quantitative detection of four typical mycotoxins (aflatoxin B1, ochratoxin A, zearalenone, and fumonisin B1) in corn samples. The test kit consisted of a nitrocellulose membrane layered with immobilized monoclonal antibodies against mycotoxins. During the assay, the mycotoxin-protein conjugates were biotinylated. The signal detection was enhanced by a combination of the biotin-streptavidin system and enhanced chemiluminescence (ECL). This improved the sensitivity of the assay. Under the optimized conditions, four calibration curves with goodness of fit (R2 > 0.98) were plotted. The results showed that the detection limits for aflatoxin B1, ochratoxin A, zearalenone, and fumonisin B1 were 0.21, 0.19, 0.09, and 0.24 ng/mL, with detection ranges of 0.47–55.69, 0.48–127.11, 0.22–31.36, and 0.56–92.57 ng/mL, respectively. The limit of detection (LOD) of this antibody microarray for aflatoxin B1, ochratoxin A, zearalenone, and fumonisin B1 in corn was 5.25, 4.75, 2.25, and 6 μg/kg, respectively. The recovery rates from the spiked samples were between 79.2% and 113.4%, with coefficient of variation <10%. The results of the analysis of commercial samples for mycotoxins using this new assay and the liquid chromatography-tandem mass spectrometry (LC-MS/MS) were comparable and in good agreement. This assay could also be modified for the simultaneous detection of other multiple mycotoxins, as well as low-weight analytes, hazardous to human health. View Full-Text
Keywords: mycotoxins; biotin-streptavidin; microarray; quantification mycotoxins; biotin-streptavidin; microarray; quantification
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Zhang, X.; Wang, Z.; Fang, Y.; Sun, R.; Cao, T.; Paudyal, N.; Fang, W.; Song, H. Antibody Microarray Immunoassay for Simultaneous Quantification of Multiple Mycotoxins in Corn Samples. Toxins 2018, 10, 415.

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