Vyvoj enzymove imunoanalyzy nekterych vyznamnych mykotoxinu

Mycotoxins are toxic secondary metabolites that may be produced by fungi in cereal grains, animal feeds, and foods both before harvest and during storage.These compounds have a wide array of chemical structures and are produced by common field and storage fungi, including many species of Aspergillus, Penicillium and Fusarium. Mycotoxins can elicit a variety of toxic symptoms in humans and animals, ranging from gastro-enteritis to cancer. A major means of eliminating mycotoxins from human and animal food is to detect and divert contaminated raw materials from feed and food use. Immunoassay procedures have several advantages: they are sensitive, specific and allow rapid determination of a large number of samples. ELISA was developed for the detection of mycotoxins namely Aflatoxin B1, Ochratoxin A, T-2 toxin and Zearalenone. Mycotoxins are not antigenic and must be conjugated to a protein carrier before immunisation. Antigens were prepared by conjugation of Aflatoxin B1-carboxymethyl oxime, Ochratoxin A, T2 toxin-hemisuccinate and Zearalenone-carboxymethyl oxime respectively with BSA. Polyclonal antisera against the mycotoxins were produced in rabbits by an immunisation. Immunoglobulin G was prepared from the antiserum by precipitation with ammonium sulphate. The precipitate was redissolved and further purified by chromatography on a Sephadex DEAE A-50 column. Resulting immunoglobulin G was purified by cellulose immunosorbent. That suspension of cellulose with bound BSA was separated. An indirect competitive ELISA was used for mycotoxins. Samples or standards were first incubated with corresponding antibodies in the microtiner plates sensitised with mycotoxin-conjugate-BSA. The plates were washed and incubated with pig anti-rabbit immunoglobulin-horse redish peroxidase conjugate. Bound enzyme activity was measured using substrate solution. Samples of feed was ground to a fine powder and a portion was extracted with acetonitril or methanol. This approach was applicable to the laboratory ELISA and the rapid screening ELISA of mycotoxins in cereals and animal feedsAvailable from STL Prague, CZ / NTK - National Technical LibrarySIGLECZCzech Republi