Detection of Allergenic Substances in Foods by a Multiplex PCR Method

Hashimoto, Hiroyuki; Makabe, Yuhki; Hasegawa, Yasuyuki; Sajiki, Junko; Miyamoto, Fumio

doi:10.3358/shokueishi.48.132

Cited by 8 publications

(4 citation statements)

References 8 publications

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“…Both of these problems may be resolved by PCR and mass spectrometry (MS, and protein sequencing). PCR has been used extensively for the detection and the speciation between related allergens, but no single assay is available to test for all allergens [13][14][15]. Further, the selective fractionation for proteinaceous material makes PCR nonquantitative and of questionable utility when assessing exposure doses for products that include soy protein, gluten, casein, and whey.…”

Section: Introductionmentioning

confidence: 98%

Multiplex detection of food allergens and gluten

et al. 2015

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To help safeguard the food supply and detect the presence of undeclared food allergens and gluten, most producers and regulatory agencies rely on commercial test kits. Most of these are ELISAs with a few being PCR-based. These methods are very sensitive and analyte specific, requiring different assays to detect each of the different food allergens. Mass spectrometry offers an alternative approach whereby multiple allergens may be detected simultaneously. However, mass spectrometry requires expensive equipment, highly trained analysts, and several years before a quantitative approach can be achieved. Using multianalyte profiling (xMAP®) technology, a commercial multiplex test kit based on the use of established antibodies was developed for the simultaneous detection of up to 14 different food allergens plus gluten. The assay simultaneously detects crustacean seafood, egg, gluten, milk, peanut, soy, and nine tree nuts (almond, Brazil nut, cashew, coconut, hazelnut, macadamia, pine nut, pistachio, and walnut). By simultaneously performing multiple tests (typically two) for each analyte, this magnetic bead-based assay offers built-in confirmatory analyses without the need for additional resources. Twenty-five of the assays were performed on buffer extracted samples, while five were conducted on samples extracted using reduced-denatured conditions. Thus, complete analysis for all 14 allergens and gluten requires only two wells of a 96-well microtiter plate. This makes it possible to include in a single analytical run up to 48 samples. All 30 bead sets in this multiplex assay detected 5 ng/mL of food allergen and gluten with responses greater than background. In addition, 26 of the bead sets displayed signal/noise ratios of five or greater. The bead-based design makes this 30-plex assay expandable to incorporate new antibodies and capture/detector methodologies by ascribing these new detectors to any of the unassigned bead sets that are commercially available.

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Section: Introductionmentioning

confidence: 98%

Multiplex detection of food allergens and gluten

et al. 2015

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“…In one multiplex reaction they detected simultaneously peanut, hazelnut, celery, and soy and in another they detected egg, milk, almond, and sesame with a LOD of 100 ppm. Also a multiplex PCR assay was developed by Hashimoto et al [102] for the simultaneous detection of wheat, buckwheat, and peanut. This multiplex format has the advantages of saving time, costs and reducing probability of cross-contamination.…”

Section: Pcr and Real-time Pcrmentioning

confidence: 99%

Recent Advances in the Detection of Allergens in Foods

Cruz

López-Calleja

Martı́n

et al. 2017

Methods in Molecular Biology

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Food allergy is a public health issue that has significantly increased worldwide in the past decade affecting consumers' quality of life and making increasing demands on health service resources. Despite recent advances in many areas of diagnosis and treatment, our general knowledge of the basic mechanisms of the disease remained limited, i.e., not at pace with the exponential number of new cases and the explosion of the new technologies. For sensitized individuals, the only effective way to prevent allergic reactions is the strict avoidance of the offending food. For this reason, a number of regulatory bodies in several countries have recognized the importance of providing information about the presence of food allergens by enacting laws, regulations, or standards for food labeling of "priority allergens." This has resulted in the need for the development of analytical methods for protection of food-allergic consumers that should be among others highly specific, sensitive, and not influenced by the presence of the food matrix components. Several analytical approaches target either the allergen itself or a corresponding allergen marker such as peptide fragment or gene segment and have been used in the detection and quantification of allergens in food products. In this short review, some of the conventional and new methods for the detection of allergens in food are listed and briefly discussed.

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“…A multiplex PCR assay has been developed for the simultaneous detection of wheat, buckwheat, and peanut (Hashimoto et al, 2007). A distinct amplicon size for each allergen can be observed in an agarose gel.…”

Section: Multiplexingmentioning

confidence: 99%