Journal
LWT-FOOD SCIENCE AND TECHNOLOGY
Volume 114, Issue -, Pages -Publisher
ELSEVIER
DOI: 10.1016/j.lwt.2019.108378
Keywords
Taqman real-time PCR; Gluten-containing cereals; Gluten-related disorders; Commercial food products
Categories
Funding
- Comunidad de Madrid (Consejeria de Educacion) [P2018/BAA-4574]
- Ministerio de Economia, Industria y Competitividad (MINECO, Spain) [AGL2017-84316-R]
- Ministerio de Educacion, Culture y Deporte (Spain)
- MINECO
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This work reports a real-time PCR assay to specifically detect the presence of DNA from the main gluten-containing cereals wheat, barley and rye (WBR) in foodstuffs. The WBR real-time PCR uses two specific primers and a Taqman probe for amplification of a 118 bp DNA fragment common to the three target cereals on the ribosomal internal transcribed spacer (ITS) region. In-house validation of the method was performed employing three binary arrays containing different concentrations (100 000-10 mg/kg) of a wheat/barley/rye mixture in maize flour: untreated, soft heat-treated at 160 degrees C/13 min and intense heat-treated at 200 degrees C/20 min. Results showed optimal PCR amplification efficiency, reproducibility and sensitivity with an overall limit of detection of 10-50 mg/kg of the target cereals, theoretically equating to approximately 1-5 mg/kg of gluten. Applicability of the assay was further assessed through a market analysis of 220 food products by the real-time PCR assay and the official R5-based ELISA. Comparative results highlighted the ability of the proposed methodology as a highly sensitive and robust procedure to detect the presence of potentially harmful concentrations of undeclared gluten-containing cereals in some of the tested samples, supporting results of the immunological assays currently used for gluten determination in foods.
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