Validation of New ELISA Technique for Detection of Aflatoxin B1 Contamination in Food Products versus HPLC and VICAM
Toxin-contaminated foods and beverages are a major source of illness, may cause death, and have a significant negative economic impact worldwide. Aflatoxin B1 (AFB1) is a potent toxin that may induce cancer after chronic low-level exposure. This study developed a quantitative recombinant <i>Af...
| Main Authors: | , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
MDPI AG
2021-10-01
|
| Series: | Toxins |
| Subjects: | |
| Online Access: | https://www.mdpi.com/2072-6651/13/11/747 |
| _version_ | 1797508281720111104 |
|---|---|
| author | Elsayed Hafez Nourhan M. Abd El-Aziz Amira M. G. Darwish Mohamed G. Shehata Amira A. Ibrahim Asmaa M. Elframawy Ahmed N. Badr |
| author_facet | Elsayed Hafez Nourhan M. Abd El-Aziz Amira M. G. Darwish Mohamed G. Shehata Amira A. Ibrahim Asmaa M. Elframawy Ahmed N. Badr |
| author_sort | Elsayed Hafez |
| collection | DOAJ |
| description | Toxin-contaminated foods and beverages are a major source of illness, may cause death, and have a significant negative economic impact worldwide. Aflatoxin B1 (AFB1) is a potent toxin that may induce cancer after chronic low-level exposure. This study developed a quantitative recombinant <i>AflR</i> gene antiserum ELISA technique for aflatoxin B1 detection in contaminated food products. Aflatoxin B1 residuals from 36 food samples were analyzed with HPLC and VICAM. DNA was extracted from aflatoxin-contaminated samples and the <i>AflR</i> gene amplified using PCR. PCR products were purified and ligated into the pGEM-T vector. Recombinant plasmids were sequenced and transformed into competent <i>E. coli</i> (BL21). Molecular size and B-cell epitope prediction for the recombinant protein were assessed. The purified protein was used to induce the production of IgG antibodies in rabbits. Serum IgG was purified and labeled with alkaline phosphatase. Finally, indirect-ELISA was used to test the effectiveness of polyclonal antibodies for detection of aflatoxin B1 in food samples. |
| first_indexed | 2024-03-10T05:00:51Z |
| format | Article |
| id | doaj.art-20ad00f26f0c47afb749ba3cfd92d97e |
| institution | Directory Open Access Journal |
| issn | 2072-6651 |
| language | English |
| last_indexed | 2024-03-10T05:00:51Z |
| publishDate | 2021-10-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Toxins |
| spelling | doaj.art-20ad00f26f0c47afb749ba3cfd92d97e2023-11-23T01:48:24ZengMDPI AGToxins2072-66512021-10-01131174710.3390/toxins13110747Validation of New ELISA Technique for Detection of Aflatoxin B1 Contamination in Food Products versus HPLC and VICAMElsayed Hafez0Nourhan M. Abd El-Aziz1Amira M. G. Darwish2Mohamed G. Shehata3Amira A. Ibrahim4Asmaa M. Elframawy5Ahmed N. Badr6Department of Plant Protection and Biomolecular Diagnosis, Arid Lands Cultivation Research Institute (ALCRI), City of Scientific Research and Technological Applications (SRTA-City), Alexandria 21934, EgyptDepartment of Food Technology, Arid Lands Cultivation Research Institute (ALCRI), City of Scientific Research and Technological Applications (SRTA-City), Alexandria 21934, EgyptDepartment of Food Technology, Arid Lands Cultivation Research Institute (ALCRI), City of Scientific Research and Technological Applications (SRTA-City), Alexandria 21934, EgyptDepartment of Food Technology, Arid Lands Cultivation Research Institute (ALCRI), City of Scientific Research and Technological Applications (SRTA-City), Alexandria 21934, EgyptDepartment of Plant Protection and Biomolecular Diagnosis, Arid Lands Cultivation Research Institute (ALCRI), City of Scientific Research and Technological Applications (SRTA-City), Alexandria 21934, EgyptNucleic Acids Research Department, Genetic Engineering & Biotechnology Research Institute (GEBRI), City of Scientific Research and Technological Applications (SRTA-City), Alexandria 21934, EgyptFood Toxicology and Contaminants Department, National Research Centre, Dokki, Cairo 12622, EgyptToxin-contaminated foods and beverages are a major source of illness, may cause death, and have a significant negative economic impact worldwide. Aflatoxin B1 (AFB1) is a potent toxin that may induce cancer after chronic low-level exposure. This study developed a quantitative recombinant <i>AflR</i> gene antiserum ELISA technique for aflatoxin B1 detection in contaminated food products. Aflatoxin B1 residuals from 36 food samples were analyzed with HPLC and VICAM. DNA was extracted from aflatoxin-contaminated samples and the <i>AflR</i> gene amplified using PCR. PCR products were purified and ligated into the pGEM-T vector. Recombinant plasmids were sequenced and transformed into competent <i>E. coli</i> (BL21). Molecular size and B-cell epitope prediction for the recombinant protein were assessed. The purified protein was used to induce the production of IgG antibodies in rabbits. Serum IgG was purified and labeled with alkaline phosphatase. Finally, indirect-ELISA was used to test the effectiveness of polyclonal antibodies for detection of aflatoxin B1 in food samples.https://www.mdpi.com/2072-6651/13/11/747aflatoxin B1recombinant <i>AflR</i> geneVICAMHPLCI-ELISApeanut |
| spellingShingle | Elsayed Hafez Nourhan M. Abd El-Aziz Amira M. G. Darwish Mohamed G. Shehata Amira A. Ibrahim Asmaa M. Elframawy Ahmed N. Badr Validation of New ELISA Technique for Detection of Aflatoxin B1 Contamination in Food Products versus HPLC and VICAM Toxins aflatoxin B1 recombinant <i>AflR</i> gene VICAM HPLC I-ELISA peanut |
| title | Validation of New ELISA Technique for Detection of Aflatoxin B1 Contamination in Food Products versus HPLC and VICAM |
| title_full | Validation of New ELISA Technique for Detection of Aflatoxin B1 Contamination in Food Products versus HPLC and VICAM |
| title_fullStr | Validation of New ELISA Technique for Detection of Aflatoxin B1 Contamination in Food Products versus HPLC and VICAM |
| title_full_unstemmed | Validation of New ELISA Technique for Detection of Aflatoxin B1 Contamination in Food Products versus HPLC and VICAM |
| title_short | Validation of New ELISA Technique for Detection of Aflatoxin B1 Contamination in Food Products versus HPLC and VICAM |
| title_sort | validation of new elisa technique for detection of aflatoxin b1 contamination in food products versus hplc and vicam |
| topic | aflatoxin B1 recombinant <i>AflR</i> gene VICAM HPLC I-ELISA peanut |
| url | https://www.mdpi.com/2072-6651/13/11/747 |
| work_keys_str_mv | AT elsayedhafez validationofnewelisatechniquefordetectionofaflatoxinb1contaminationinfoodproductsversushplcandvicam AT nourhanmabdelaziz validationofnewelisatechniquefordetectionofaflatoxinb1contaminationinfoodproductsversushplcandvicam AT amiramgdarwish validationofnewelisatechniquefordetectionofaflatoxinb1contaminationinfoodproductsversushplcandvicam AT mohamedgshehata validationofnewelisatechniquefordetectionofaflatoxinb1contaminationinfoodproductsversushplcandvicam AT amiraaibrahim validationofnewelisatechniquefordetectionofaflatoxinb1contaminationinfoodproductsversushplcandvicam AT asmaamelframawy validationofnewelisatechniquefordetectionofaflatoxinb1contaminationinfoodproductsversushplcandvicam AT ahmednbadr validationofnewelisatechniquefordetectionofaflatoxinb1contaminationinfoodproductsversushplcandvicam |