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Toxins 2019, 11(1), 48; https://doi.org/10.3390/toxins11010048

Development of Indirect Competitive Enzyme-Linked Immunosorbent Assay to Detect Fusarium verticillioides in Poultry Feed Samples

1
Department of Pathological Sciences, State University of Londrina, P.O. box 10.011, Londrina 86057-970, Paraná, Brazil
2
Department of Biochemistry and Biotechnology, State University of Londrina, P.O. box 10.011, Londrina 86057-970, Paraná, Brazil
3
Department of Food Science and Technology, State University of Londrina, P.O. box 10.011, Londrina 86057-970, Paraná, Brazil
4
Department of General Biology, State University of Londrina, P.O. box 10.011, Londrina 86057-970, Paraná, Brazil
*
Author to whom correspondence should be addressed.
Received: 14 December 2018 / Revised: 10 January 2019 / Accepted: 11 January 2019 / Published: 17 January 2019
(This article belongs to the Special Issue Advanced Methods for Mycotoxins Detection)
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Abstract

Fumonisins are a group of toxic secondary metabolites that are produced by Fusarium verticillioides which are associated with poultry health hazard and great economic losses. The objective of the present study was to develop an immunological method to detect F. verticillioides in poultry feed samples. An indirect competitive enzyme-linked immunosorbent assay (ic-ELISA) based on a polyclonal antibody against 67 kDa protein of the F. verticillioides 97K exoantigen was developed to detect this fungus. Antibody anti-67 kDa protein showed cross-reactivity against F. graminearum (2–7%) and F. sporotrichioides (10%), but no or low cross-reactivity against Aspergillus sp. and Penicillium sp. exoantigens. The detection limit for the 67 kDa protein of F. verticillioides was 29 ng/mL. Eighty-one poultry feed samples were analyzed for Fusarium sp. count, 67 kDa protein of F. verticillioides and fumonisin concentrations. Eighty of the 81 feed samples (98.6%) showed Fusarium sp. contamination (mean 6.2 x 104 CFU/g). Mean 67 kDa protein and fumonisin concentration in the poultry feed samples was 21.0 µg/g and 1.02 µg/g, respectively. The concentration of 67 kDa protein, as determined by ic-ELISA correlated positively (p < 0.05) with fumonisin levels (r = 0.76). These results suggest that this ic-ELISA has potential to detect F. verticillioides and predict fumonisin contamination in poultry feed samples. View Full-Text
Keywords: Toxigenic fungi; polyclonal antibodies; immunoassay; mycotoxin; feed safety; food-borne fungi Toxigenic fungi; polyclonal antibodies; immunoassay; mycotoxin; feed safety; food-borne fungi
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This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited (CC BY 4.0).
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Omori, A.M.; Ono, E.Y.S.; Hirozawa, M.T.; de Souza Suguiura, I.M.; Hirooka, E.Y.; Pelegrinelli Fungaro, M.H.; Ono, M.A. Development of Indirect Competitive Enzyme-Linked Immunosorbent Assay to Detect Fusarium verticillioides in Poultry Feed Samples. Toxins 2019, 11, 48.

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