Person:
Cuadrado Vives, María Carmen

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First Name
María Carmen
Last Name
Cuadrado Vives
URI
https://hdl.handle.net/20.500.14352/77721
Affiliation
Universidad Complutense de Madrid
Faculty / Institute
Farmacia
Department
Nutrición y Ciencia de los Alimentos
Area
Nutrición y Bromatología
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2016 - 20182
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Author: Linacero De La Fuente, M. Rosario × Subject: 664 ×

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Now showing 1 - 2 of 2
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    Evaluation of locked nucleic acid and TaqMan probes for specific detection of cashew nut in processed food by real time PCR
    (Food control, 2018) Sanchiz Giraldo, África; Ballesteros Redondo, María Isabel; Marqués, Eric; Diéguez, Carmen; Rueda Muñoz de San Pedro, Julia; Cuadrado Vives, María Carmen; Linacero De La Fuente, M. Rosario
    Cashew (Anacardium occidentale) nut can trigger serious reactions in allergic patients, including anaphylaxis and death. Labelling the presence of cashew nuts in food products is mandatory and consequently, sensitive and specific analytical methods must be developed. In this study, Ana o allergen coding sequences have been sequenced in several cashew varieties. Two hydrolysis probes, locked nucleic acid (LNA) and TaqMan, have been designed and their efficiency, sensitivity, limit of detection and specificity for Ana o 1 coding-sequence detection have been compared. Reliable Real Time PCR assays to detect and quantify up to 10 ppm of cashew nuts in complex mixtures have been developed. Moreover, the influence of boiling and autoclave treatment on cashew nut detectability has been analysed by qPCR, showing both probes similar performance. This analytical method was able to detect up to 1000 ppm with good functionality in autoclave treated samples. Boiling did not affect cashew nut detectability. Both hydrolysis probes are suitable for Ana o 1 coding sequence detection. Applicability of the assay has been studied by analysing several food products, and comparing the results with those of a commercial ELISA kit.
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    Detection by real time PCR of walnut allergen coding sequences in processed foods
    (Food Chemistry, 2016) 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
    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.