Analytical Validation of a Direct Competitive ELISA for Multiple Mycotoxin Detection in Human Serum
Mycotoxin exposure in humans is primarily assessed through its occurrence in external sources, such as food commodities. Herein, we have developed a direct competitive ELISA to facilitate the detection of aflatoxin B1 (AFB1), deoxynivalenol (DON), fumonisin (FUM B1/B2), ochratoxin A (OTA), and zeara...
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| Format: | Article |
| Language: | English |
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MDPI AG
2022-10-01
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| Series: | Toxins |
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| Online Access: | https://www.mdpi.com/2072-6651/14/11/727 |
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| author | Kunal Garg Fausto Villavicencio-Aguilar Flora Solano-Rivera Leona Gilbert |
| author_facet | Kunal Garg Fausto Villavicencio-Aguilar Flora Solano-Rivera Leona Gilbert |
| author_sort | Kunal Garg |
| collection | DOAJ |
| description | Mycotoxin exposure in humans is primarily assessed through its occurrence in external sources, such as food commodities. Herein, we have developed a direct competitive ELISA to facilitate the detection of aflatoxin B1 (AFB1), deoxynivalenol (DON), fumonisin (FUM B1/B2), ochratoxin A (OTA), and zearalenone (ZEA) in human serum. The analytical validation of the assay followed practices endorsed by the international research community and the EU directive 96/23/EC in order to examine detection capability, recovery, and cross-reactivity. The assay demonstrated a lower limit of quantitation (LLOQ) for AFB1 [0.61 ng/mL (hereon ng/mL = ppb)], DON (19.53 ppb), FUM (4.88 ppb), OTA (19.53 ppb), and ZEA (0.15 ppb). Recovery from human serum for all mycotoxins spanned from 73% to 106%. Likewise, the specificity for monoclonal antibodies against cross-reactant mycotoxins ranged from 2% to 11%. This study compares the LLOQ and recovery values with commercial and emerging immuno-based methods for detecting mycotoxins in foodstuffs. The LLOQ values from the present study were among the lowest in commercial or emerging methods. Despite the differences in the extraction protocols and matrices, the recovery range in this study, commercial tests, and other procedures were similar for all mycotoxins. Overall, the assay detected AFB1, DON, FUM, OTA, and ZEA in human serum with excellent accuracy, precision, and specificity. |
| first_indexed | 2024-03-09T18:35:03Z |
| format | Article |
| id | doaj.art-3c5f66e89f1c40909f75971a31ad51e3 |
| institution | Directory Open Access Journal |
| issn | 2072-6651 |
| language | English |
| last_indexed | 2024-03-09T18:35:03Z |
| publishDate | 2022-10-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Toxins |
| spelling | doaj.art-3c5f66e89f1c40909f75971a31ad51e32023-11-24T07:10:15ZengMDPI AGToxins2072-66512022-10-01141172710.3390/toxins14110727Analytical Validation of a Direct Competitive ELISA for Multiple Mycotoxin Detection in Human SerumKunal Garg0Fausto Villavicencio-Aguilar1Flora Solano-Rivera2Leona Gilbert3Tezted Ltd., Mattilaniemi 6-8, 40100 Jyväskylä, FinlandSanoviv Medical Institute, KM 39 Carretera Libre Tijuana-Ensenada s/n Interior 6, Playas de Rosarito, Baja 11 California, Rosarito 22710, MexicoSanoviv Medical Institute, KM 39 Carretera Libre Tijuana-Ensenada s/n Interior 6, Playas de Rosarito, Baja 11 California, Rosarito 22710, MexicoTezted Ltd., Mattilaniemi 6-8, 40100 Jyväskylä, FinlandMycotoxin exposure in humans is primarily assessed through its occurrence in external sources, such as food commodities. Herein, we have developed a direct competitive ELISA to facilitate the detection of aflatoxin B1 (AFB1), deoxynivalenol (DON), fumonisin (FUM B1/B2), ochratoxin A (OTA), and zearalenone (ZEA) in human serum. The analytical validation of the assay followed practices endorsed by the international research community and the EU directive 96/23/EC in order to examine detection capability, recovery, and cross-reactivity. The assay demonstrated a lower limit of quantitation (LLOQ) for AFB1 [0.61 ng/mL (hereon ng/mL = ppb)], DON (19.53 ppb), FUM (4.88 ppb), OTA (19.53 ppb), and ZEA (0.15 ppb). Recovery from human serum for all mycotoxins spanned from 73% to 106%. Likewise, the specificity for monoclonal antibodies against cross-reactant mycotoxins ranged from 2% to 11%. This study compares the LLOQ and recovery values with commercial and emerging immuno-based methods for detecting mycotoxins in foodstuffs. The LLOQ values from the present study were among the lowest in commercial or emerging methods. Despite the differences in the extraction protocols and matrices, the recovery range in this study, commercial tests, and other procedures were similar for all mycotoxins. Overall, the assay detected AFB1, DON, FUM, OTA, and ZEA in human serum with excellent accuracy, precision, and specificity.https://www.mdpi.com/2072-6651/14/11/727human biological monitoringmultiple mycotoxinsenzyme-linked immunosorbent assay |
| spellingShingle | Kunal Garg Fausto Villavicencio-Aguilar Flora Solano-Rivera Leona Gilbert Analytical Validation of a Direct Competitive ELISA for Multiple Mycotoxin Detection in Human Serum Toxins human biological monitoring multiple mycotoxins enzyme-linked immunosorbent assay |
| title | Analytical Validation of a Direct Competitive ELISA for Multiple Mycotoxin Detection in Human Serum |
| title_full | Analytical Validation of a Direct Competitive ELISA for Multiple Mycotoxin Detection in Human Serum |
| title_fullStr | Analytical Validation of a Direct Competitive ELISA for Multiple Mycotoxin Detection in Human Serum |
| title_full_unstemmed | Analytical Validation of a Direct Competitive ELISA for Multiple Mycotoxin Detection in Human Serum |
| title_short | Analytical Validation of a Direct Competitive ELISA for Multiple Mycotoxin Detection in Human Serum |
| title_sort | analytical validation of a direct competitive elisa for multiple mycotoxin detection in human serum |
| topic | human biological monitoring multiple mycotoxins enzyme-linked immunosorbent assay |
| url | https://www.mdpi.com/2072-6651/14/11/727 |
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