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Multi-component immunochromatographic assay for simultaneous detection of aflatoxin B1, ochratoxin A and zearalenone in agro-food.

Biosensors & bioelectronics (2013-06-29)
Xin Li, Peiwu Li, Qi Zhang, Ran Li, Wen Zhang, Zhaowei Zhang, Xiaoxia Ding, Xiaoqian Tang
ABSTRACT

Mycotoxins are highly toxic contaminants and have induced health threat to human and animals. Aflatoxin B1 (AFB1), ochratoxin A (OTA) and zearalenone (ZEA) commonly occur in food and feed. A multi-component immunochromatographic assay (ICA) was developed for rapid and simultaneous determination of these three mycotoxins in agro-food. The strategy was performed based on the competitive immunoreactions between antibody-colloidal gold nanoparticle conjugate probes and mycotoxins or mycotixin antigens. Each monoclonal antibody specially recognize its corresponding mycotoxin and antigen, and there was no cross reactivity in the assay. Three mycotixin antigens were immobilized as three test lines in the nitrocellulose membrane reaction zone, which enable the simultaneous detection in one single test. The visible ICA results were obtained in 20 min. The visual detection limits of this strip test for the AFB1, OTA and ZEA were 0.25 ng/mL, 0.5 ng/mL and 1 ng/mL, respectively. The assay was evaluated using spiked and naturally contaminated peanuts, maize and rice samples. The results were in accordance with those obtained using enzyme-linked immunosorbent assay. In summary, this developed ICA could provide an effective and rapid approach for onsite detection of multi-mycotoxin in agro-food samples without any expensive instrument.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Aflatoxin B1 from Aspergillus flavus, from Aspergillus flavus
Supelco
Ochratoxin A solution, 10 μg/mL in acetonitrile, analytical standard
Sigma-Aldrich
Ochratoxin A, from Petromyces albertensis, ≥98% (HPLC)
Sigma-Aldrich
Zearalenone, fungal mycotoxin
Supelco
Aflatoxin B1 solution, 2 μg/mL in acetonitrile, analytical standard
Supelco
Ochratoxin A, from Aspergillus ochraceus, reference material
Supelco
Zearalenone solution, 100 μg/mL in acetonitrile, analytical standard
Supelco
Zearalenone, reference material
Aflatoxin B1 solution, 3.79 μg/g in acetonitrile, ERM®, certified reference material