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, clone 2G6 produced a monoclonal antibody that had equal sensitivity to aflatoxins G1 and G2, and did not cross-react with aflatoxins B1, B2, or M1. Its IC50 values for aflatoxins G1 and G2 were 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 also used both methods to determine the amounts of G-group aflatoxins in five peanut samples contaminated by aflatoxin B1-positive, and their relative standard deviations ranged from 8.4% to 17.7% (under 20%), which demonstrates a good correlation between the two methods. We further used this CI-ELISA to assess the ability of 126 fungal strains isolated from peanuts or field 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 2 G6-based CI-ELISA was suitable for the detection of G-group aflatoxins present in peanuts and also those produced by fungi.
Highlights
Aflatoxins, a group of highly toxic and carcinogenic difuranocoumarin compounds, are secondary metabolites produced by Aspergillus flavus and Aspergillus parasiticus [1]
HPLC method, which gave recovery rates ranging from 92% to 102%. These results demonstrate that the developed CI-enzyme-linked immunosorbent assay (ELISA) is suitable for the detection of G-group aflatoxins in peanuts
We examined a total of 126 putative A. flavus strains previously isolated from peanuts or field soil samples by the CI-ELISA method to determine if some isolates are A. parasiticus
Summary
Aflatoxins, a group of highly toxic and carcinogenic difuranocoumarin compounds, are secondary metabolites produced by Aspergillus flavus and Aspergillus parasiticus [1]. Toxins 2016, 8, 5; doi:10.3390/toxins8010005 www.mdpi.com/journal/toxins aflatoxins identified, those belonging to the two major groups, B and G (Figure 1), are frequently found found to contaminate food and feed, especially and maize [2,3,4]. To their extremely to contaminate food and feed, especially peanuts peanuts and maize. Due to their toxic and toxic and carcinogenic effects, legal limits have been imposed on various agri‐food products in carcinogenic effects, legal limits have been imposed on various agri-food products in different countries different countries and regions ofexample, the worldthe [5].Codex. −1 for total aflatoxins [6]. 1 foroftotal Joint FAO/WHO
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