Development Of Phage Enzyme-linked Immunosorbent Assay For Detecting Deoxynivalenol And Preliminary Study On Immunoassay Of Patulin

Mycotoxin Deoxynivalenol (DON) is a kind of trichothecenes produced by Fusarium. Although DON is one of the least acutely toxic trichothecenes, it should be treated as an important food safety issue, because it is a common contaminant of wheat and corn, usually occurs in temperate climates in years that are particularly wet at the time of harvest. DON causes feed refusal, emesis, immunosuppression or immunostimulation, teratogeny, cytotoxicity, reproductive toxicity and human cancers. Mycotoxin patulin (PAT) is a toxic secondary metabolite produced by a wide range of fungal species of Penicillium and Aspergillus, and is considered to be mutagenic, carcinogenic, teratogenic and immunologic toxicity. Therefore, a rapid and reliable method for the determination of deoxynivalenol and patulin is important to avoid the risk of DON consumption by humans and animals. Some methods have been developed for detecting deoxynivalenol and patulin in food products, such as Thin-layer chromatography (TLC), High-performace liquid chromatography (HPLC) and Gas chromatography (GC), but these methods that provide qualitative results are considerably expensive and time consuming owing to the use of large amounts of organic solvents, complicated clean-up procedures and expensive equipments, inconvenient for routine assay. Enzyme-linked immunosorbent assays (ELISA) employing monoclonal or polyclonal antibodies is by far the most widely used serological test for the detection of mycotoxins because of its simplicity, rapidity, and sensitivity, and ELISA has a unique ability to routinely handle a large number of samples and don't require time-consuming procedures and sophisticated equipments. In this study, the 3-hemiglutarate-deoxynivalenol (3-HG-DON) was prepared by protection of the C7- and C15- hydroxyls with a cyclic boronate ester, esterification with glutarate at the C3- hydroxyl, and removal of the boronate ester. 3-HG-DON was activated by active ester method, and conjugated to ovalbumin (OVA) as the DON detecting antigen (DON-HG-OVA). DON was activated by...