Application of an enzyme-linked immunosorbent assay for detecting mold contamination in agricultural commodities and comparison with conventional assays

Abstract

Mold contamination in agricultural commodities including grains, grain-based foods, fruits, and vegetables was investigated by two analytical methods. One method employed a mixed monoclonal antibody sandwich ELISA for detection of Aspergillus, Penicillium and Fusarium genera-specific antigens. The detection limit of ELISA for these antigens was 100 ng/g. The other protocol was HPLC-based detection of ergosterol, the predominant sterol in most molds. Recoveries of ergosterol from grains and grain-based foods at spiking levels of 50 Á/1000 ng g (1 ranged between 82 and 86%. The detection limit of the HPLC method for ergosterol was 40 ng g (1 . Ninety-two samples (59%) and 116 (75%) among 155 samples tested positive with antigens and ergosterol by the ELISA and the HPLC method, respectively. The cause of conflicting data between both detection methods found in nine dried persimmon samples were solved by using the conventional direct plating method that showed the prevalence of other genera with rarity of Aspergillus, Penicillium and Fusarium. The sandwich ELISA method offers a rapid assay for selectively tracing the potential mycotoxigenic mold, and also is a preliminary screening method for mycotoxin analysis.

Fungal microflora in dried persimmon fruits

“…In only one sample, yeast count was found above 5.48 log cfu/g. In comparison to the results obtained in this study, Park et al (2003) obtained lower levels of mould count in dried persimmon fruits that ranged between 2.0 and 3.26 log cfu/g.…”

“…Moulds are common contaminants of agricultural products. Mould contamination in food not only results in spoilage or economic losses but is a serious concern for human and animal health since it may produce toxic metabolites called mycotoxins (Park et al, 2003). Mycotoxin formation can take place at any of the stages of growing, harvesting, transporting, drying and during the storage of dried fruit and vegetable products.…”

Development and Application of an Immunoaffinity Column Enzyme Immunoassay for Mycotoxin Zearalenone in Complicated Samples

“…It is well known that sensitivity and specificity were the most important parameters for the antibodies and for their assay methods [23] . The IC 50 value was the main criterion to evaluate the sensitivity of the mAbs, and the cross-reactivity revealed the specificity of the mAbs [24] . The titer results, IC 50 results for each clone in Table 2 indicated that the titer of 2D3 (1.5×10 5 ) was the highest among them.…”

Detection ofFusariumsp. contamination in corn by enzyme-linked immunosorbent assay

“…The development of the enzyme-linked immunosorbent assay (ELISA) (Engvall & Pearlmann 1972) could be a promising alternative to the traditional methods (Lin et al 1986). Several ELISAs have been developed for detecting and identifying moulds with variable success (Notermans et al 1986, Tsai & Yu 1997, Chen et al 2000, Yong & Cousin 2001, Eibel et al 2005, Yucel et al 2005, however a limited number have been developed for the detection of Fusarium species in foods (Gan et al 1997, Abramson et al 1998, Iyer & Cousin 2003, Park et al 2003.…”