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Development of a multiplex flow cytometric microsphere immunoassay for mycotoxins and evaluation of its application in feed
A multi-mycotoxin immunoassay—using the MultiAnalyte Profiling (xMAP) technology—is developed and evaluated. This technology combines a unique color-coded microsphere suspension array, with a dedicated flow cytometer. We aimed for the combined detection of aflatoxins, ochratoxin A, deoxynivalenol, f...
Autores principales: | , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Springer-Verlag
2010
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3150826/ https://www.ncbi.nlm.nih.gov/pubmed/21836765 http://dx.doi.org/10.1007/s12550-010-0077-0 |
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author | Peters, Jeroen Bienenmann-Ploum, Monique de Rijk, Theo Haasnoot, Willem |
author_facet | Peters, Jeroen Bienenmann-Ploum, Monique de Rijk, Theo Haasnoot, Willem |
author_sort | Peters, Jeroen |
collection | PubMed |
description | A multi-mycotoxin immunoassay—using the MultiAnalyte Profiling (xMAP) technology—is developed and evaluated. This technology combines a unique color-coded microsphere suspension array, with a dedicated flow cytometer. We aimed for the combined detection of aflatoxins, ochratoxin A, deoxynivalenol, fumonisins, zearalenone and T-2-toxin in an inhibition immunoassay format. Sets of six mycotoxin-protein conjugates and six specific monoclonal antibodies were selected, and we observed good sensitivities and no cross-interactions between the assays in buffer. However, detrimental effects of the feed extract on the sensitivities and in some cases on the slopes of the curves were observed and different sample materials showed different effects. Therefore, for quantitative analysis, this assay depends on calibration curves in blank matrix extracts or on the use of a suitable multi-mycotoxin cleanup. To test if the method was suitable for the qualitative detection at EU guidance levels, we fortified rapeseed meal, a feed ingredient, with the six mycotoxins, and all extracts showed inhibited responses in comparison with the non-fortified sample extract. Contaminated FAPAS reference feed samples assigned for a single mycotoxin showed strong inhibitions in the corresponding assays but also often in other assays of the multiplex. In most cases, the presence of these other mycotoxins was confirmed by instrumental analysis. The multiplex immunoassay can be easily extended with other mycotoxins of interest, but finding a suitable multi-mycotoxin cleanup will improve its applicability. |
format | Online Article Text |
id | pubmed-3150826 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2010 |
publisher | Springer-Verlag |
record_format | MEDLINE/PubMed |
spelling | pubmed-31508262011-08-09 Development of a multiplex flow cytometric microsphere immunoassay for mycotoxins and evaluation of its application in feed Peters, Jeroen Bienenmann-Ploum, Monique de Rijk, Theo Haasnoot, Willem Mycotoxin Res Original Paper A multi-mycotoxin immunoassay—using the MultiAnalyte Profiling (xMAP) technology—is developed and evaluated. This technology combines a unique color-coded microsphere suspension array, with a dedicated flow cytometer. We aimed for the combined detection of aflatoxins, ochratoxin A, deoxynivalenol, fumonisins, zearalenone and T-2-toxin in an inhibition immunoassay format. Sets of six mycotoxin-protein conjugates and six specific monoclonal antibodies were selected, and we observed good sensitivities and no cross-interactions between the assays in buffer. However, detrimental effects of the feed extract on the sensitivities and in some cases on the slopes of the curves were observed and different sample materials showed different effects. Therefore, for quantitative analysis, this assay depends on calibration curves in blank matrix extracts or on the use of a suitable multi-mycotoxin cleanup. To test if the method was suitable for the qualitative detection at EU guidance levels, we fortified rapeseed meal, a feed ingredient, with the six mycotoxins, and all extracts showed inhibited responses in comparison with the non-fortified sample extract. Contaminated FAPAS reference feed samples assigned for a single mycotoxin showed strong inhibitions in the corresponding assays but also often in other assays of the multiplex. In most cases, the presence of these other mycotoxins was confirmed by instrumental analysis. The multiplex immunoassay can be easily extended with other mycotoxins of interest, but finding a suitable multi-mycotoxin cleanup will improve its applicability. Springer-Verlag 2010-11-26 2011 /pmc/articles/PMC3150826/ /pubmed/21836765 http://dx.doi.org/10.1007/s12550-010-0077-0 Text en © The Author(s) 2010 https://creativecommons.org/licenses/by-nc/4.0/ This article is distributed under the terms of the Creative Commons Attribution Noncommercial License which permits any noncommercial use, distribution, and reproduction in any medium, provided the original author(s) and source are credited. |
spellingShingle | Original Paper Peters, Jeroen Bienenmann-Ploum, Monique de Rijk, Theo Haasnoot, Willem Development of a multiplex flow cytometric microsphere immunoassay for mycotoxins and evaluation of its application in feed |
title | Development of a multiplex flow cytometric microsphere immunoassay for mycotoxins and evaluation of its application in feed |
title_full | Development of a multiplex flow cytometric microsphere immunoassay for mycotoxins and evaluation of its application in feed |
title_fullStr | Development of a multiplex flow cytometric microsphere immunoassay for mycotoxins and evaluation of its application in feed |
title_full_unstemmed | Development of a multiplex flow cytometric microsphere immunoassay for mycotoxins and evaluation of its application in feed |
title_short | Development of a multiplex flow cytometric microsphere immunoassay for mycotoxins and evaluation of its application in feed |
title_sort | development of a multiplex flow cytometric microsphere immunoassay for mycotoxins and evaluation of its application in feed |
topic | Original Paper |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3150826/ https://www.ncbi.nlm.nih.gov/pubmed/21836765 http://dx.doi.org/10.1007/s12550-010-0077-0 |
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