A sensitive and specific real-time PCR targeting DNA from wheat, barley and rye to track gluten contamination in marketed foods

García-García, Aina; Madrid, Raquel; Sohrabi, Hamidreza; Cruz, Silvia de la; Garcı́a, Teresa; Martı́n, Rosario; González, Isabel

doi:10.1016/j.lwt.2019.108378

Cited by 21 publications

(9 citation statements)

References 26 publications

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“…At present, enzyme-linked immunosorbent assay (ELISA) is the most commonly used method for gluten analysis not only for its selectivity and sensitivity but also for the lack of other reference methods (García-García et al, 2020;Lacorn et al, 2019;Morón et al, 2008;Sajic et al, 2020;Sherf, 2017;Skerrit and Hill, 1990;Valdés et al, 2003;Zhanga et al, 2019). Besides ELISA, other analytical methodologies are currently being developed like mass spectrometry combined with proteomic technologies (Colgrave et al, 2015;Fiedler et al, 2018), methods based on DNA amplification (García-García et al, 2019;Garrido-Maestu et al, 2018;Martín-Fernández et al, 2015 and electrochemical sensors using as recognition elements molecular imprinted polymers (Iskierko et al, 2019) and aptamers (López-López et al, 2017;Malbano et al, 2017;White et al, 2018). Among electrochemical sensors, aptasensors are emerging as alternatives to reference methods.…”

Section: Introductionmentioning

confidence: 99%

Truncated aptamers as selective receptors in a gluten sensor supporting direct measurement in a deep eutectic solvent

Svigelj

Dossi

Pizzolato

et al. 2020

Biosensors and Bioelectronics

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Section: Introductionmentioning

confidence: 99%

Truncated aptamers as selective receptors in a gluten sensor supporting direct measurement in a deep eutectic solvent

Svigelj

Dossi

Pizzolato

et al. 2020

Biosensors and Bioelectronics

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“…The Spanish National Center for Plant Genetic Resources (CRF-INIA, Madrid, Spain) kindly supplied different kernels of common, spelt, rivet and durum wheats, barley and rye (10 cultivars each). More information about these reference samples can be obtained from García-García et al 2019 [ 27 ]. Non-target plant species included in the specificity assays and 30 commercially available foodstuffs from several brands were acquired from local retailers and food stores in Madrid (Spain) including products with different declarations with regard to the gluten-containing cereals.…”

Section: Methodsmentioning

confidence: 99%

Production of a Recombinant Single-Domain Antibody for Gluten Detection in Foods Using the Pichia pastoris Expression System

García-García

Madrid

Garcia-Calvo

et al. 2020

Foods

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The detection of gluten in foodstuffs has become a growing concern in food allergen management as a result of the high ratio of population sensitive to the main gluten-containing cereals. In this study, a promising single-domain antibody previously isolated by phage display (dAb8E) was produced in Pichia pastoris resulting in high levels of the antibody fragment expression (330 mg/L). The purified dAb8E was proved to specifically bind to gluten proteins from wheat, barley and rye, exhibiting no cross reaction to other heterologous species. The dynamic range of the sandwich enzyme-linked immunosorbent assay (ELISA) covered 0.1 to 10 µg/mL of gliadin, reaching a limit of detection of 0.12 µg/mL. When experimental binary mixtures of the target cereals were analyzed, the limit of detection was 0.13 mg/g, which would theoretically correspond to gluten concentrations of approximately 13 mg/kg. Finally, thirty commercially available food products were analyzed by means of the developed assay to further confirm the applicability of the dAb8E for gluten determination. The proposed methodology enabled the generation of a new gluten-specific nanobody which could be used to guarantee the appropriate labelling of gluten-free foods.

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“…Real time PCR methods can be also used for the quantitative detection of gluten when using model mixtures [ 46 , 47 ]. Moreover, PCR methods perform indirect detection (not pathogenic compounds of gluten—rather the DNA that codifies for it) [ 48 ].…”

Section: Methods For Gluten Detection In Food Samplesmentioning

confidence: 99%

From Polyclonal Sera to Recombinant Antibodies: A Review of Immunological Detection of Gluten in Foodstuff

Garcia-Calvo

García-García

Madrid

et al. 2020

Foods

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Gluten is the ethanol-soluble protein fraction of cereal endosperms like wheat, rye, and barley. It is widely used in the food industry because of the physical–chemical properties it gives to dough. Nevertheless, there are some gluten-related diseases that are presenting increasing prevalences, e.g., celiac disease, for which a strict gluten-free diet is the best treatment. Due to this situation, gluten labeling legislation has been developed in several countries around the world. This article reviews the gluten immune detection systems that have been applied to comply with such regulations. These systems have followed the development of antibody biotechnology, which comprise three major methodologies: polyclonal antibodies, monoclonal antibodies (mAbs) derived from hybridoma cells (some examples are 401.21, R5, G12, and α-20 antibodies), and the most recent methodology of recombinant antibodies. Initially, the main objective was the consecution of new high-affinity antibodies, resulting in low detection and quantification limits that are mainly achieved with the R5 mAb (the gold standard for gluten detection). Increasing knowledge about the causes of gluten-related diseases has increased the complexity of research in this field, with current efforts not only focusing on the development of more specific and sensitive systems for gluten but also the detection of protein motifs related to pathogenicity. New tools based on recombinant antibodies will provide adequate safety and traceability methodologies to meet the increasing market demand for gluten-free products.

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A sensitive and specific real-time PCR targeting DNA from wheat, barley and rye to track gluten contamination in marketed foods

Cited by 21 publications

References 26 publications

Truncated aptamers as selective receptors in a gluten sensor supporting direct measurement in a deep eutectic solvent

Truncated aptamers as selective receptors in a gluten sensor supporting direct measurement in a deep eutectic solvent

Production of a Recombinant Single-Domain Antibody for Gluten Detection in Foods Using the Pichia pastoris Expression System

From Polyclonal Sera to Recombinant Antibodies: A Review of Immunological Detection of Gluten in Foodstuff

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