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ARS Home » Southeast Area » New Orleans, Louisiana » Southern Regional Research Center » Food and Feed Safety Research » Research » Publications at this Location » Publication #323926

Research Project: Control of Aflatoxin Production by Targeting Aflatoxin Biosynthesis

Location: Food and Feed Safety Research

Title: Development of an enzyme-linked immunosorbent assay method specific for the detection of G-group aflatoxins

Author
item LI, PEIWU - Oil Crops Research Institute - China
item ZHOU, QIAN - Oil Crops Research Institute - China
item WANG, TING - Oil Crops Research Institute - China
item ZHOU, HAIYAN - Oil Crops Research Institute - China
item ZHANG, WEN - Oil Crops Research Institute - China
item DING, XIAOXIA - Oil Crops Research Institute - China
item ZHANG, ZHAOWEI - Oil Crops Research Institute - China
item Chang, Perng Kuang
item ZHANG, QI - Oil Crops Research Institute - China

Submitted to: Toxins
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 12/11/2015
Publication Date: 1/1/2016
Citation: Li, P., Zhou, Q., Wang, T., Zhou, H., Zhang, W., Ding, X., Zhang, Z., Chang, P.-K., Zhang, Q. 2016. Development of an enzyme-linked immunosorbent assay method specific for the detection of G-group aflatoxins. Toxins. 8:5. doi: 10.3390/toxins8010005.

Interpretive Summary: Aflatoxins, a group of highly toxic and carcinogenic compounds, are produced by Aspergillus species. Of all aflatoxins identified, those belonging to B and G groups are frequently found to contaminate agricultural commodities. Although the toxicity of G-group aflatoxins is lower than that of aflatoxin B1, there is still a need to monitor this toxin group in food and feed to protect health of humans and animals. To this end, we generated a antibody that recognizes aflatoxins G1 and G2 and does not cross-react with aflatoxins B1 and B2. With this novel antibody, a sensitive immune assay was developed.

Technical Abstract: To detect and monitor G-group aflatoxins in agricultural products, we generated class-specific monoclonal antibodies that specifically recognized aflatoxins G1 and G2. Of the final three positive and stable hybridomas obtained, hybridoma 2G6 produced a monoclonal antibody that did not cross-react with aflatoxins B1, B2 or M1 but had equal sensitivity to aflatoxins G1 and G2. Its IC50 for aflatoxins G1 and G2 was 17.18 ng mL-1 and 19.75 ng mL-1, respectively. Using this new monoclonal antibody, we developed a competitive indirect enzyme-linked immunosorbent assay (CI-ELISA); the method had a limit of detection of 0.06 ng mL-1. To validate this CI-ELISA, we spiked uncontaminated peanut samples with various amounts of aflatoxins G1 and G2 and compared recovery rates with those determined by a standard HPLC method. The recovery rates of the CI-ELISA ranging from 94% to 103% were comparable to those of the HPLC (92% to 102%). We further used this CI-ELISA to assess the ability of 126 fungal strains isolated from peanuts or soils to produce G-group aflatoxins. Among these, seven stains producing different amounts of G-group aflatoxins were identified. Our results showed that the monoclonal antibody 2G6-based CI-ELISA was suitable for the detection of G-group aflatoxins present in peanuts and also those produced by fungi.