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Anal Chem. 2017 Nov 7;89(21):11520-11528. doi: 10.1021/acs.analchem.7b02794. Epub 2017 Oct 11.

Time-Resolved Fluorescence Immunochromatographic Assay Developed Using Two Idiotypic Nanobodies for Rapid, Quantitative, and Simultaneous Detection of Aflatoxin and Zearalenone in Maize and Its Products.

Tang X1,2,3,4,5, Li P1,2,3,4,5, Zhang Q4, Zhang Z3, Zhang W1,2,3, Jiang J1.

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Oil Crops Research Institute, Chinese Academy of Agricultural Sciences , Wuhan 430062, China.
Key Laboratory of Biology and Genetic Improvement of Oil Crops, Ministry of Agriculture , Wuhan 430062, China.
Laboratory of Quality & Safety Risk Assessment for Oilseed Products (Wuhan), Ministry of Agriculture , Wuhan 430062, China.
Key Laboratory of Detection for Mycotoxins, Ministry of Agriculture , Wuhan 430062, China.
Quality Inspection & Test Center for Oilseed Products, Ministry of Agriculture , Wuhan 430062, China.


Aflatoxins and zearalenone (ZEN) are highly common mycotoxins in maize and maize-based products. This study aimed to report a time-resolved fluorescence immunochromatographic assay (TRFICA) developed using two idiotypic nanobodies for rapid, quantitative, and simultaneous detection of aflatoxin B1 (AFB1) and ZEN in maize and its products. A novel Eu/Tb(III) nanosphere with enhanced fluorescence was prepared as a label and conjugated to anti-idiotypic nanobody (AIdnb) and monoclonal antibody (mAb). On the basis of nanosphere-antibody conjugation, two patterns of competitive time-resolved strip methods (AIdnb-TRFICA and mAb-TRFICA) were established and compared. The half inhibition concentration of AIdnb-TRFICA was 0.46 and 0.86 ng·mL-1 for AFB1 and ZEN, which was 18.3- and 20.3-fold more sensitive than that of mAb-TRFICA for AFB1 and ZEN, respectively. Under optimal conditions, AIdnb-TRFICA for dual mycotoxin was established and provided a quantitative relationship ranging from 0.13 to 4.54 ng·mL-1 for AFB1 and 0.20 to 2.77 ng·mL-1 for ZEN, with a detection limit of 0.05 and 0.07 ng·mL-1 in the buffer solution, respectively. AIdnb-TRFICA showed good recoveries (72.6%-106.6%) in samples and was applied to detect dual mycotoxin in maize samples with satisfying results. To the best of our knowledge, it is the first report about a time-resolved strip method based on AIdnbs for dual mycotoxin.

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