Detection of Genetically Modified Corn (Bt176) in Spiked Cow Blood Samples by Polymerase Chain Reaction and Immunoassay Methods
| Autor: | Yves Bertheau, Stephen Lamart, Patrick Fach, André Kobilinsky, Marie-Noëlle Duplan, Patrice Martin, Laetitia Petit, K. Duhem, P. Brunschwig, Alexandra Schattner, Annick Diolez, Fabienne Baraige |
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| Přispěvatelé: | Agence Française de Sécurité Sanitaire des Aliments (AFSSA), Unité de recherche Phytopathologie et Méthodologies de la Détection (PMDV), Institut National de la Recherche Agronomique (INRA), Institut de l'élevage (IDELE), Centre National Interprofessionnel de l'Economie Laitière (CNIEL), Unité de biométrie et intelligence artificielle de jouy, Arilait Recherches, Unité de recherche génomique et physiologie de la lactation (GPL) |
| Rok vydání: | 2005 |
| Předmět: |
Chloroplasts
[SDV]Life Sciences [q-bio] Food Genetically Modified 01 natural sciences Analytical Chemistry law.invention Hemolysin Proteins chemistry.chemical_compound law [MATH]Mathematics [math] Promoter Regions Genetic IMMUNOASSAY METHOD Polymerase Polymerase chain reaction 2. Zero hunger Genetics biology medicine.diagnostic_test Reverse Transcriptase Polymerase Chain Reaction 04 agricultural and veterinary sciences Plants 040401 food science Genetically modified organism DNA Plant GENETICALLY MODIFIED CORN Transgene Bacterial Toxins Enzyme-Linked Immunosorbent Assay Zea mays 0404 agricultural biotechnology Bacterial Proteins medicine Animals Environmental Chemistry [INFO]Computer Science [cs] Gene DNA Primers Cell Nucleus Pharmacology Bacillus thuringiensis Toxins 010401 analytical chemistry DNA extraction Molecular biology 0104 chemical sciences Endotoxins Bt176 chemistry Immunoassay biology.protein Cattle Agronomy and Crop Science DNA Food Science |
| Zdroj: | HAL Journal of AOAC INTERNATIONAL Journal of AOAC INTERNATIONAL, AOAC International, 2005, 88 (2), pp.654-664 Scopus-Elsevier |
| ISSN: | 1944-7922 1060-3271 |
| DOI: | 10.1093/jaoac/88.2.654 |
| Popis: | The fate of DNA and protein transgenic sequences in products derived from animals fed transgenic crops has recently raised public interest. Sensitive molecular tests targeting the Bt176 genetic construct and the transgenic Cry1Ab protein were developed to determine whether plant sequences, especially transgenic sequences, are present in animal products. A protocol for total DNA extraction and purification from cow whole blood samples was first drawn up and assessed by spiking with known amounts of DNA from Bt176 maize. The limit of detection for transgenic sequences (35S promoter and Bt176-specific junction sequence) was determined by both the polymerase chain reaction–enzyme-linked immunosorbent assay (PCR–ELISA) and the 5′-nuclease PCR assay. Four additional PCR systems were built to substantiate the results. The first detects a mono-copy maize-specific sequence (ADH promoter). Two others target multi-copy sequences from plant nucleus (26S rRNA gene) and chloroplast (psaB gene). The last one, used as a positive control, targets a mono-copy animal sequence (αs1-casein gene). Both methods detected a minimum spiking at 25 copies of Bt176 maize/mL in 10 mL whole blood samples. The sandwich ELISA kit used detected down to 1 ng transgenic Cry1Ab protein/mL spiked whole blood. |
| Databáze: | OpenAIRE |
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