Detection by real time PCR of walnut allergen coding sequences in processed foods
A quantitative real-time PCR (RT-PCR) method, employing novel primer sets designed on Jug r 1, Jug r 3, and Jug r 4 allergen-coding sequences, was set up and validated. Its specificity, sensitivity, and applicability were evaluated. The DNA extraction method based on CTAB–phenol–chloroform was best...
| Autores: | ; ; ; ; ; ; ; ; ; ; ; |
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| Tipo de documento: | Artículo |
| Fecha de publicación: | 2016 |
| País: | España |
| Institución: | Universidad Complutense de Madrid (UCM) |
| Repositorio: | Docta Complutense |
| Lenguaje: | Inglés |
| OAI Identifier: | oai:docta.ucm.es:20.500.14352/23364 |
| Acceso en línea: | https://hdl.handle.net/20.500.14352/23364 |
| Nivel de acceso: | Acceso abierto |
| Palabra clave: | 664 579.67 Juglans regia Walnut allergen detection Real-time PCR Processed foods Thermal processing Pressure processing Biotecnología Genética 3399 Otras Especialidades Tecnológicas 2409 Genética |
| _version_ | 1878742156117540864 |
|---|---|
| author | Linacero De La Fuente, M. Rosario |
| author2 | Ballesteros Redondo, María Isabel Sanchiz Giraldo, África Prieto, Nuria Iniesto, Elisa Martínez, Yolanda Martín Pedrosa, Mercedes Muzquiz, Mercedes Cabanillas Martín, Beatriz Rovira, Mercè Burbano, Carmen Cuadrado Vives, María Carmen |
| author2_role | author author author author author author author author author author author |
| author_browse | Ballesteros Redondo, María Isabel Burbano, Carmen Cabanillas Martín, Beatriz Cuadrado Vives, María Carmen Iniesto, Elisa Linacero De La Fuente, M. Rosario Martín Pedrosa, Mercedes Martínez, Yolanda Muzquiz, Mercedes Prieto, Nuria Rovira, Mercè Sanchiz Giraldo, África |
| author_facet | Linacero De La Fuente, M. Rosario Ballesteros Redondo, María Isabel Sanchiz Giraldo, África Prieto, Nuria Iniesto, Elisa Martínez, Yolanda Martín Pedrosa, Mercedes Muzquiz, Mercedes Cabanillas Martín, Beatriz Rovira, Mercè Burbano, Carmen Cuadrado Vives, María Carmen |
| author_role | author |
| collection | Docta Complutense |
| description | A quantitative real-time PCR (RT-PCR) method, employing novel primer sets designed on Jug r 1, Jug r 3, and Jug r 4 allergen-coding sequences, was set up and validated. Its specificity, sensitivity, and applicability were evaluated. The DNA extraction method based on CTAB–phenol–chloroform was best for walnut. RT-PCR allowed a specific and accurate amplification of allergen sequence, and the limit of detection was 2.5 pg of walnut DNA. The method sensitivity and robustness were confirmed with spiked samples, and Jug r 3 primers detected up to 100 mg/kg of raw walnut (LOD 0.01%, LOQ 0.05%). Thermal treatment combined with pressure (autoclaving) reduced yield and amplification (integrity and quality) of walnut DNA. High hydrostatic pressure (HHP) did not produce any effect on the walnut DNA amplification. This RT-PCR method showed greater sensitivity and reliability in the detection of walnut traces in commercial foodstuffs compared with ELISA assays. |
| eu_rights_str_mv | openAccess |
| format | article |
| id | ES_f10d9a582c4c796af6acb1e2a362daa2 |
| institution | Universidad Complutense de Madrid (UCM) |
| instname_str | Universidad Complutense de Madrid (UCM) |
| language | eng |
| network_acronym_str | ES |
| network_name_str | España |
| oai_identifier_str | oai:docta.ucm.es:20.500.14352/23364 |
| publishDate | 2016 |
| publishDateSort | 2016 |
| publisher | Elsevier |
| publisherStr | Elsevier |
| reponame_str | Docta Complutense |
| spelling | Detection by real time PCR of walnut allergen coding sequences in processed foods Linacero De La Fuente, M. Rosario Ballesteros Redondo, María Isabel Sanchiz Giraldo, África Prieto, Nuria Iniesto, Elisa Martínez, Yolanda Martín Pedrosa, Mercedes Muzquiz, Mercedes Cabanillas Martín, Beatriz Rovira, Mercè Burbano, Carmen Cuadrado Vives, María Carmen 664 579.67 Juglans regia Walnut allergen detection Real-time PCR Processed foods Thermal processing Pressure processing Biotecnología Genética 3399 Otras Especialidades Tecnológicas 2409 Genética A quantitative real-time PCR (RT-PCR) method, employing novel primer sets designed on Jug r 1, Jug r 3, and Jug r 4 allergen-coding sequences, was set up and validated. Its specificity, sensitivity, and applicability were evaluated. The DNA extraction method based on CTAB–phenol–chloroform was best for walnut. RT-PCR allowed a specific and accurate amplification of allergen sequence, and the limit of detection was 2.5 pg of walnut DNA. The method sensitivity and robustness were confirmed with spiked samples, and Jug r 3 primers detected up to 100 mg/kg of raw walnut (LOD 0.01%, LOQ 0.05%). Thermal treatment combined with pressure (autoclaving) reduced yield and amplification (integrity and quality) of walnut DNA. High hydrostatic pressure (HHP) did not produce any effect on the walnut DNA amplification. This RT-PCR method showed greater sensitivity and reliability in the detection of walnut traces in commercial foodstuffs compared with ELISA assays. Elsevier https://hdl.handle.net/20.500.14352/23364 |
| spellingShingle | Detection by real time PCR of walnut allergen coding sequences in processed foods Linacero De La Fuente, M. Rosario 664 579.67 Juglans regia Walnut allergen detection Real-time PCR Processed foods Thermal processing Pressure processing Biotecnología Genética 3399 Otras Especialidades Tecnológicas 2409 Genética |
| title | Detection by real time PCR of walnut allergen coding sequences in processed foods |
| title_full | Detection by real time PCR of walnut allergen coding sequences in processed foods |
| title_fullStr | Detection by real time PCR of walnut allergen coding sequences in processed foods |
| title_full_unstemmed | Detection by real time PCR of walnut allergen coding sequences in processed foods |
| title_short | Detection by real time PCR of walnut allergen coding sequences in processed foods |
| title_sort | Detection by real time PCR of walnut allergen coding sequences in processed foods |
| topic | 664 579.67 Juglans regia Walnut allergen detection Real-time PCR Processed foods Thermal processing Pressure processing Biotecnología Genética 3399 Otras Especialidades Tecnológicas 2409 Genética |
| topic_facet | 664 579.67 Juglans regia Walnut allergen detection Real-time PCR Processed foods Thermal processing Pressure processing Biotecnología Genética 3399 Otras Especialidades Tecnológicas 2409 Genética |
| url | https://hdl.handle.net/20.500.14352/23364 |