Ring trial validation of single and multiplex real-time PCR methods for the detection and quantification of the allergenic food ingredients mustard, celery, soy, wheat and rye

Waiblinger, Hans‐Ulrich; Boernsen, Britta; Geppert, Carina; Demmel, Anja; Peterseil, Verena; Koeppel, René

doi:10.1007/s00003-016-1063-z

Cited by 12 publications

(4 citation statements)

References 14 publications

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“…A certain increase of S R, rel and S r, rel could be expected, if the DNA extraction step was performed individually by each lab. In previous validation studies, DNA extraction had been included (Siegel et al 2013;Waiblinger et al 2014Waiblinger et al , 2017, thus experience to what extent the individual DNA preparation leads to additional uncertainty is already available. Within the collaboration trial, the performance of the PCR method had to be evaluated.…”

Section: Precisionmentioning

confidence: 99%

Collaborative trial validation of a new multiplex real-time PCR to sensitively detect allergenic nuts in food

Waiblinger

Geppert

Bartsch

et al. 2022

J Consum Prot Food Saf

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In this article, we present a multiplex real-time PCR method for a simultaneous, sensitive and specific detection and semi-quantitative estimation of the allergenic species peanut, hazelnut, walnut and cashew in food. Due to the use of multicopy target sequences, a very sensitive detection of the allergenic ingredients was possible. The method was validated in-house as well as by a collaborative trial with 12 laboratories. Within the ring trial, 0.64 mg/kg (i.e. approx. 0.1–0.2 mg of peanut and tree nut-derived protein/kg) could still be detected in a processed cookie matrix, confirmed by results of incurred, processed samples spiked at very low levels between 0.9 and 50 mg/kg of the corresponding allergenic ingredient (peanut, tree nut). In addition, the method revealed good precision data. With regard to quantitative analysis though, insufficient recovery data (bias) were determined in some cases, resulting in measurement uncertainties of more than 50%.

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Section: Precisionmentioning

confidence: 99%

Collaborative trial validation of a new multiplex real-time PCR to sensitively detect allergenic nuts in food

Waiblinger

Geppert

Bartsch

et al. 2022

J Consum Prot Food Saf

Self Cite

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“…Conversely, the presence of a natural matrix induces thermal instability mainly due to physical (i.e., hydrophobic effect) and chemical interactions (i.e., thiol-disulfide interchange) compromising the extractability and immunodetection. Therefore, even with the same food sample, the matrix components can show variations depending on the protocol used to prepare the extract, thus affecting the results in terms of allergen profile and concentration [ 33 ].…”

Section: Introductionmentioning

confidence: 99%

Detection of Allergenic Proteins in Foodstuffs: Advantages of the Innovative Multiplex Allergen Microarray-Based Immunoassay Compared to Conventional Methods

Tuppo

Giangrieco

Tamburrini

et al. 2022

Foods

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Several factors can affect the allergen content and profile of a specific food, including processing procedures often leading to a decrease in allergenicity, although no change, or even an increase, have also been reported. Evaluation of the effectiveness of a processing procedure requires the availability of reliable methodologies to assess the variation in molecules able to induce allergic reactions in the analyzed food. Conventional and innovative strategies and methodologies can be exploited to identify allergenic proteins in foodstuffs. However, depending on the specific purposes, different methods can be used. In this review, we have critically reviewed the advantages of an innovative method, the multiplex allergen microarray-based immunoassay, in the detection of allergens in foodstuffs. In particular, we have analyzed some studies reporting the exploitation of an IgE-binding inhibition assay on multiplex allergen biochips, which has not yet been reviewed in the available literature. Unlike the others, this methodology enables the identification of many allergenic proteins, some of which are still unknown, which are recognized by IgE from allergic patients, with a single test. The examined literature suggests that the inhibition test associated with the multiplex allergen immunoassay is a promising methodology exploitable for the detection of IgE-binding proteins in food samples.

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“…The most straightforward idea is absolute quantification, where a serial dilution of the target DNA sequence is used as the standard curve [7]. Thereby, the template can be either directly isolated DNA of the pure target [8], a plasmid containing the cloned DNA sequence [9], or synthetically synthesized DNA [10]. This quantification method is well suited for raw food samples.…”

Section: Introductionmentioning

confidence: 99%

Comparison of Real-Time PCR Quantification Methods in the Identification of Poultry Species in Meat Products

Dolch

Andrée

Schwägele

2020

Foods

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Poultry meat is consumed worldwide and is prone to food fraud because of large price differences among meat from different poultry species. Precise and sensitive analytical methods are necessary to control poultry meat products. We chose species–specific sequences of the cytochrome b gene to develop two multiplex real-time polymerase chain reaction (real-time PCR) systems: one for chicken (Gallus gallus), guinea fowl (Numida meleagris), and pheasant (Phasianus colchicus), and one for quail (Coturnix japonica) and turkey (Meleagris gallopavo). For each species, added meat could be detected down to 0.5 % w/w. No cross reactions were seen. For these two real-time PCR systems, we applied three different quantification methods: (A) with relative standard curves, (B) with matrix-specific multiplication factors, and (C) with an internal DNA reference sequence to normalize and to control inhibition. All three quantification methods had reasonable recovery rates from 43% to 173%. Method B had more accepted recovery rates, i.e., in the range 70–130%, namely 83% compared to 75% for method A or C.

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Ring trial validation of single and multiplex real-time PCR methods for the detection and quantification of the allergenic food ingredients mustard, celery, soy, wheat and rye

Cited by 12 publications

References 14 publications

Collaborative trial validation of a new multiplex real-time PCR to sensitively detect allergenic nuts in food

Collaborative trial validation of a new multiplex real-time PCR to sensitively detect allergenic nuts in food

Detection of Allergenic Proteins in Foodstuffs: Advantages of the Innovative Multiplex Allergen Microarray-Based Immunoassay Compared to Conventional Methods

Comparison of Real-Time PCR Quantification Methods in the Identification of Poultry Species in Meat Products

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