Validating allergen coding genes (Cor a 1, Cor a 8, Cor a 14) as target sequences for hazelnut detection via Real-Time PCR

D’Andrea, Matteo; Coı̈sson, Jean Daniel; Locatelli, Monica; Garino, Cristiano; Cereti, Elisabetta

doi:10.1016/j.foodchem.2010.07.029

Cited by 18 publications

(12 citation statements)

References 28 publications

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“…These primers were checked for their specificity utilizing BLAST and employed for the amplifications of partial allergen-coding sequences of three almond cultivars. This procedure makes it possible to assess the intraspecific polymorphism which could produce false negatives . All the amplicons showed the expected size, and their sequences were homologous to those employed for designing the specific primers (data in Table 1 of the Supporting Information).…”

Section: Resultsmentioning

confidence: 99%

Detection of Almond Allergen Coding Sequences in Processed Foods by Real Time PCR

Prieto

Iniesto

Burbano³

et al. 2014

J. Agric. Food Chem.

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The aim of this work was to develop and analytically validate a quantitative RT-PCR method, using novel primer sets designed on Pru du 1, Pru du 3, Pru du 4, and Pru du 6 allergen-coding sequences, and contrast the sensitivity and specificity of these probes. The temperature and/or pressure processing influence on the ability to detect these almond allergen targets was also analyzed. All primers allowed a specific and accurate amplification of these sequences. The specificity was assessed by amplifying DNA from almond, different Prunus species and other common plant food ingredients. The detection limit was 1 ppm in unprocessed almond kernels. The method's robustness and sensitivity were confirmed using spiked samples. Thermal treatment under pressure (autoclave) reduced yield and amplificability of almond DNA; however, high-hydrostatic pressure treatments did not produced such effects. Compared with ELISA assay outcomes, this RT-PCR showed higher sensitivity to detect almond traces in commercial foodstuffs.

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Section: Resultsmentioning

confidence: 99%

Detection of Almond Allergen Coding Sequences in Processed Foods by Real Time PCR

Prieto

Iniesto

Burbano³

et al. 2014

J. Agric. Food Chem.

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“…In terms of absolute LOD, the conventional real-time PCR system tested in this work allowed tracing 5 pg of diluted hazelnut DNA that was lower than the LOD of 9.6 pg and 13 pg of hazelnut reported. 17,21 The proposed nested real-time PCR system was able to increase the sensitivity by 1 order of magnitude, allowing the detection of 0.5 pg of hazelnut DNA. To our knowledge, this is the lowest LOD ever reported in the literature for the absolute detection of hazelnut DNA.…”

Section: ■ Results and Discussionmentioning

confidence: 99%

“…[17][18][19]21 However, to validate and correctly implement this and/or other methodologies for monitoring the presence of allergens in foods, it is essential that certified reference materials are developed, such as those in the case of GMO. Official guidelines should also be made available shortly, regulating limits for the presence of potentially allergenic ingredients in food and recommending methodology for their monitoring.…”

Section: ■ Results and Discussionmentioning

confidence: 99%

Single-Tube Nested Real-Time PCR as a New Highly Sensitive Approach to Trace Hazelnut

Costa

Kuchta

Oliveira

2012

J. Agric. Food Chem.

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Hazelnut is one of the most commonly consumed tree nuts, being largely used by the food industry in a wide variety of processed foods. However, it is a source of allergens capable of inducing mild to severe allergic reactions in sensitized individuals. Hence, the development of highly sensitive methodologies for hazelnut traceability is essential. In this work, we developed a novel technique for hazelnut detection based on a single-tube nested real-time PCR system. The system presents high specificity and sensitivity, enabling a relative limit of detection of 50 mg/kg of hazelnut in wheat material and an absolute limit of detection of 0.5 pg of hazelnut DNA (1 DNA copy). Its application to processed food samples was successfully achieved, detecting trace amounts of hazelnut in chocolate down to 60 mg/kg. These results highlight the adequacy of the technique for the specific detection and semiquantitation of hazelnut as potential hidden allergens in foods.

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“…Recombinant plasmids containing species‐specific target sequences can be obtained to serve as an internal plasmid reference standard (IPRS). IPRS has been exploited to detect allergens in food . In conjunction with SYBR Green I, they have been used to quantify materials contaminated by almonds ( Prunus dulcis ) …”

Section: Introductionmentioning

confidence: 99%

Detection of allergen coding sequences of kiwi, peach, and apple in processed food by qPCR

2018

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Validating allergen coding genes (Cor a 1, Cor a 8, Cor a 14) as target sequences for hazelnut detection via Real-Time PCR

Cited by 18 publications

References 28 publications

Detection of Almond Allergen Coding Sequences in Processed Foods by Real Time PCR

Detection of Almond Allergen Coding Sequences in Processed Foods by Real Time PCR

Single-Tube Nested Real-Time PCR as a New Highly Sensitive Approach to Trace Hazelnut

Detection of allergen coding sequences of kiwi, peach, and apple in processed food by qPCR

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