Matrix-normalised real-time PCR approach to quantify soybean as a potential food allergen as affected by thermal processing

Tree nuts show nutritional properties and human health benefits. However, they contain allergenic proteins, which make them harmful to the sensitised population. The presence of tree nuts on food labelling is mandatory and, consequently, the development of suitable analytical methodologies to detect nuts in processed foods is advisable. Real-Time PCR allowed a specific and accurate amplification of allergen sequences. Some food processing methods could induce structural and/or conformational changes in proteins by altering their allergenic capacity, as well as produce the fragmentation and/or degradation of genomic DNA. In this work, we analysed by means of Real-Time PCR, the influence of pressure and thermal processing through Instant Controlled Pressure Drop (DIC) on the detectability of hazelnut, pistachio and cashew allergens. The detection of targets in hazelnut, pistachio and cashew (Cor a 9, Pis v 1 and Ana o 1, respectively) is affected by the treatment to different extents depending on the tree nut. Results are compared to those previously obtained by our group in the analysis of different treatments on the amplificability of the same targets. Reduction in amplificability is similar to that reported for some autoclave conditions. Our assays might allow for the detection of up to 1000 mg/kg of hazelnut, pistachio and cashew flours after being submitted to DIC treatment in food matrices.

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Effect of Instant Controlled Pressure Drop (DIC) Treatment on the Detection of Nuts by Real Time PCR

10.20944/preprints202005.0076.v1 ◽

2020 ◽

Author(s):

Africa Sanchiz ◽

Carmen Cuadrado ◽

Joseph Haddad ◽

Rosario Linacero

Keyword(s):

Pressure Drop ◽

Real Time ◽

Conformational Changes ◽

Real Time Pcr ◽

Food Processing ◽

Thermal Processing ◽

Tree Nuts ◽

Allergenic Proteins ◽

Instant Controlled Pressure Drop ◽

Sensitive Population

Tree nuts show nutritional properties and human health benefits. However, they contain allergenic proteins, which make them harmful to the sensitive population. The presence of tree nuts on food labelling is mandatory and, consequently, the development of suitable analytical methodologies to detect nuts in processed foods is advisable. Real-time PCR allowed a specific and accurate amplification of allergen sequences. Some food processing methods could induce structural and/or conformational changes in proteins by altering their allergenic capacity, as well as produce the fragmentation and/or degradation of genomic DNA. In this work, we analysed by means of Real-time PCR, the influence of pressure and thermal processing through Instant Controlled Pressure Drop (DIC) on the detectability of hazelnut; pistachio and cashew allergens have been tested. The detection of targets in hazelnut, pistachio and cashew (Cor a 9, Pis v 1 and Ana o 1, respectively) is affected by the treatment, in different extent depending on the tree nut. Results are compared to those previously obtained by our group in the analysis of different treatments on the amplificability of the same targets. Reduction in amplificability is similar to that reported for some autoclave conditions. Our assays might allow detecting up to 1000 ppm of hazelnut, pistachio and cashew flours after being submitted to DIC treatment in food matrices

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Detection of Lupine (Lupinus spp. L.) as a food allergen using three methods: end-point PCR, Real-Time PCR and Elisa

Potravinarstvo Slovak Journal of Food Sciences ◽

10.5219/384 ◽

2014 ◽

Vol 8 (1) ◽

Author(s):

Ondrej Revák ◽

Jozef Golian ◽

Radoslav Židek ◽

Jozef Čapla ◽

Peter Zajác

Keyword(s):

Real Time ◽

Food Allergen ◽

Real Time Pcr ◽

End Point

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Effect of food matrix and thermal processing on the performance of a normalised quantitative real-time PCR approach for lupine (Lupinus albus) detection as a potential allergenic food

Food Chemistry ◽

10.1016/j.foodchem.2018.04.079 ◽

2018 ◽

Vol 262 ◽

pp. 251-259 ◽

Cited By ~ 13

Author(s):

Caterina Villa ◽

Joana Costa ◽

Cristina Gondar ◽

M. Beatriz P.P. Oliveira ◽

Isabel Mafra

Keyword(s):

Real Time ◽

Real Time Pcr ◽

Thermal Processing ◽

Lupinus Albus ◽

Allergenic Food ◽

Food Matrix ◽

Quantitative Real Time Pcr ◽

Effect Of Food

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Effect of Instant Controlled Pressure Drop (DIC) Treatment on the Detection of Nut Allergens by Real Time PCR

10.20944/preprints202005.0076.v2 ◽

2020 ◽

Author(s):

Africa Sanchiz ◽

Carmen Cuadrado ◽

Joseph Haddad ◽

Rosario Linacero

Keyword(s):

Pressure Drop ◽

Real Time ◽

Conformational Changes ◽

Real Time Pcr ◽

Food Processing ◽

Thermal Processing ◽

Nutritional Properties ◽

Tree Nuts ◽

Allergenic Proteins ◽

Instant Controlled Pressure Drop

Tree nuts show nutritional properties and human health benefits. However, they contain allergenic proteins, which make them harmful to the sensitised population. The presence of tree nuts on food labelling is mandatory and, consequently, the development of suitable analytical methodologies to detect nuts in processed foods is advisable. Real-time PCR allowed a specific and accurate amplification of allergen sequences. Some food processing methods could induce structural and/or conformational changes in proteins by altering their allergenic capacity, as well as produce the fragmentation and/or degradation of genomic DNA. In this work, we analysed by means of Real-time PCR, the influence of pressure and thermal processing through Instant Controlled Pressure Drop (DIC) on the detectability of hazelnut,pistachio and cashew allergens have been tested. The detection of targets in hazelnut, pistachio and cashew (Cor a 9, Pis v 1 and Ana o 1, respectively) is affected by the treatment, in different extent depending on the tree nut. Results are compared to those previously obtained by our group in the analysis of different treatments on the amplificability of the same targets. Reduction in amplificability is similar to that reported for some autoclave conditions. Our assays might allow detecting up to 1000 mg/kg of hazelnut, pistachio and cashew flours after being submitted to DIC treatment in food matrices.

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A Novel PCR Method for Quantification of Buckwheat by Using a Unique Internal Standard Material

Journal of Food Protection ◽

10.4315/0362-028x-69.10.2478 ◽

2006 ◽

Vol 69 (10) ◽

pp. 2478-2486 ◽

Cited By ~ 19

Author(s):

TAKASHI HIRAO ◽

MASAYUKI HIRAMOTO ◽

SHINSUKE IMAI ◽

HISANORI KATO

Keyword(s):

Real Time ◽

Dna Extraction ◽

Food Allergen ◽

Real Time Pcr ◽

Internal Standard ◽

Food Samples ◽

Internal Transcribed Spacer Region ◽

Standard Material ◽

Wild Buckwheat ◽

Pcr Targeting

A novel quantitative and specific method for detection of buckwheat, a known food allergen, in diverse food materials was developed by using a unique internal standard to compensate for the variability in DNA extraction and amplification efficiencies. The method was based on a real-time PCR targeting the internal transcribed spacer region of Fagopyrum spp. and was designed to detect both cultivated and wild buckwheat, because wild buckwheat might be potentially allergenic. As the internal standard material, ground seeds of statice (Limonium sinuatum) were added to food samples prior to DNA extraction, and the amount of statice DNA measured by real-time PCR was used to standardize the buckwheat content. Statice, an ornamental plant, was chosen as the internal standard material because it was readily available and was inferred to be least likely to be commingled in foods. The specificity of the PCR system was tested against commonly used food materials of plant origin. Quantitative results expressed in buckwheat protein concentrations (mean ± standard deviation) for various food samples prepared to contain 10 ppm (wt/wt) of buckwheat flour (corresponding to 1.2-μg/g [ppm] buckwheat protein) ranged from 0.7 ± 0.2 (rice) to 0.9 ± 0.4 (wheat) and for 100-ppm (wt/wt) samples (12-μg/g [ppm] buckwheat protein) from 7.7 ± 1.0 (pepper) to 9.8 ± 0.5 (wheat) μg/g (ppm). The method's accuracy, sensitivity, and specificity were considered sufficient for detection of buckwheat contamination at the level required for compliance with the Japanese Food Allergen Labeling Regulation.

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