Digital PCR for analysis of peanut and soybean allergens in foods

Pierboni, Elisa; Rondini, Cristina; Torricelli, Martina; Ciccone, Letizia; Tovo, Gloria Raquel; Mercuri, Maria Lucia; Altissimi, Serena; Haouet, Naceur

doi:10.1016/j.foodcont.2018.04.039

Cited by 23 publications

(11 citation statements)

References 35 publications

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“…Different commercial ELISA kits with varying sensitivity toward quantification are available in the market to assay peanut allergens (Jayasena et al., ); hence, depending on the food matrix being tested, the detection should be selected with utmost care. Moreover in recent years, genomic approaches (such as PCR; López‐Calleja et al., ; Pierboni et al., ; Scaravelli, Brohée, Marchelli, & Van Hengel, ; Watanabe et al., ) and proteomic‐based approaches (such as mass spectrometry [MS]; Boo, Parker, & Jackson, ; Chassaigne, Nørgaard, & Van Hengel, ; Wei, Gledhill, & Maleki, ) has gained prominence to detect peanut allergen components. DNA‐based approaches may have an advantage over ELISA as the detection is based on expressed DNA, which has a higher stability in food processing methods, particularly since processing may influence the extraction efficiency of proteins, and may affect the quality of protein analysis methods due to alterations in allergens (Pierboni et al., ; Scaravelli et al., ).…”

Section: Peanut Allergy: Mechanism Clinical Relevance and Current Mmentioning

confidence: 99%

“…Moreover in recent years, genomic approaches (such as PCR; López‐Calleja et al., ; Pierboni et al., ; Scaravelli, Brohée, Marchelli, & Van Hengel, ; Watanabe et al., ) and proteomic‐based approaches (such as mass spectrometry [MS]; Boo, Parker, & Jackson, ; Chassaigne, Nørgaard, & Van Hengel, ; Wei, Gledhill, & Maleki, ) has gained prominence to detect peanut allergen components. DNA‐based approaches may have an advantage over ELISA as the detection is based on expressed DNA, which has a higher stability in food processing methods, particularly since processing may influence the extraction efficiency of proteins, and may affect the quality of protein analysis methods due to alterations in allergens (Pierboni et al., ; Scaravelli et al., ). On the contrary, genomic approaches to detect DNA components beside the allergenic proteins may also be challenging if the DNA is altered by such processing, and the detection is based on the source DNA material rather than its allergenic protein that directly represent its IgE binding and allergenicity.…”

Section: Peanut Allergy: Mechanism Clinical Relevance and Current Mmentioning

confidence: 99%

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Peanut Allergy: Characteristics and Approaches for Mitigation

Shah

Shi

Ashley

et al. 2019

Comp Rev Food Sci Food Safe

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Peanut allergy has garnered significant attention because of the high sensitization rate, increase in allergy, and severity of the reaction. Sufficiently reliable therapies and efficient mitigating techniques to combat peanut allergy are still lacking. Current management relies on avoiding peanuts and nuts and seeds with homologous proteins, although adverse events mostly occur with accidental ingestion. There is a need for hypoallergenic peanut products to protect sensitized individuals and perhaps serve as immunotherapeutic products. Alongside traditional practices of thermal and chemical treatment, novel processing approaches such as high‐pressure processing, pulsed ultraviolet light, high‐intensity ultrasound, irradiation, and pulsed electric field have been performed toward reducing the immunoreactivity of peanut. Covalent and noncovalent chemical modifications to proteins also have the tendency to alter peanut allergenicity. Enzymatic hydrolysis seems to be the most advantageous technique in diminishing the allergenic potential of peanut. Furthermore, the combined processing approach (hurdle technologies) such as enzymatic hydrolysis followed by, or in conjunction with, roasting, high pressure and heat, ultrasound with enzymatic treatment, or germination have shown a significant reduction of peanut immunoreactivity and may emerge as useful techniques in reducing the allergenicity of peanut and other foods. This study represents our current knowledge about the alterations in allergenic properties of peanut via different processing mechanisms as well as evaluating its future potential, geographical based data on increasing sensitization, clinical relevance, eliciting dose, and current management of peanut allergy. Furthermore, the molecular characteristics and clinical relevance of peanut allergens have been discussed.

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Section: Peanut Allergy: Mechanism Clinical Relevance and Current Mmentioning

confidence: 99%

Section: Peanut Allergy: Mechanism Clinical Relevance and Current Mmentioning

confidence: 99%

Peanut Allergy: Characteristics and Approaches for Mitigation

Shah

Shi

Ashley

et al. 2019

Comp Rev Food Sci Food Safe

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“…Futhermore, digital PCR, the last generation of PCR, was applied for the detection of peanut and soybean in foods (Pierboni et al., 2018). This method is based on fractioned PCR through partitioning a sample into many droplets or cavity for separate reactions and partitioned counting positive and negative.…”

Section: Real‐time Fluorescence Quantification Pcrmentioning

confidence: 99%

Fluorescence‐based quantitative platform for ultrasensitive food allergen detection: From immunoassays to DNA sensors

Qian

Zhou

et al. 2020

Comp Rev Food Sci Food Safe

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Food allergies are global health issue with an increasing prevalence that affect food safety; hence, food allergen detection, labeling, and management are considered to be important priorities in the food industry. In this critical review, we provide a comprehensive overview of several fluorescence-based platforms based on different biorecognition ligands, such as antibodies, DNA, aptamers, and cells, for food allergen quantification. Traditional analytical methods are generally unsuitable for food manufacturers to accomplish the real-time identification of food allergens in food products. Therefore, it is important to develop simple, rapid, inexpensive, accurate, and sensitive methods to improve user accessibility. A fluorescence-based quantitative platform provides an excellent detection platform for food allergens because of its high sensitivity. This review summarizes the traditional antibody-based fluorescent techniques for food allergen detection, such as the time-resolved fluoroimmunoassay , immunofluorescence imaging, fluorescence enzyme-linked immune sorbent assay, flow injection fluoroimmunoassay, and fluorescence immunosensors. However, these methods suffer from disadvantages such as the significant rate of false-positive and false-negative results due to antibody cross-reactivity with nontarget food components in the complex food matrix and epitope degradation during food processing. Hence, different types of fluorescence-based immunoassays are suitable for standardization and quantification of allergens in fresh foods. In addition, we summarize new fluorescence-based quantitative platforms, including fluorescence genosensors, fluorescence cell sensors, and fluorescence aptamer sensors. With the advantages of high sensitivity and simple operation, fluorescence biosensors will have great potential in the future and could provide portable methods for multiallergen real-time detection in complex food systems.

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“…The PCR, especially real-time PCR such as the TaqMan assay, is a very specific and sensitive tool. The specificity and sensitivity are due to the use of specific primers and probes that, in allergen analysis, recognise and amplify the allergen coding region or other constitutive genes of the target as an indirect method [ 18 ].…”

Section: Introductionmentioning

confidence: 99%

“…The aim of the study was to implement the panel of food allergens already investigated in our control laboratory (soybean and peanut) [ 18 ]. Therefore, the main objective was to identify systems that worked well with the same annealing temperature to carry out a multi-analysis, adopting a TaqMan chemistry with the same thermal profile, a unique reaction mix, and an optimised preparatory phase in order to detect low and specific quantities of allergens with a detailed and extensive investigation among related species.…”

Section: Introductionmentioning

confidence: 99%

Sesame, Pistachio, and Macadamia Nut: Development and Validation of New Allergenic Systems for Fast Real-Time PCR Application

Torricelli

Pierboni

Rondini

et al. 2020

Foods

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Food allergy is a worldwide health problem that concerns infants to adults. The main health risk for sensitised individuals is due to the presence of traces of allergens as the result of an accidental contamination during food processing. The labelling of allergens such as sesame, pistachio, and macadamia nut on food products is mandatory according to Regulation (EU) N. 1169/2011; therefore, the development of suitable and specific analytical methodologies is advisable. The aim of this study was to perform a multi-allergen real-time PCR system that works well in fast mode at the same annealing temperature and with the same thermal profile. The real-time PCR was developed designing new, specific, and efficient primer and probe systems for the 2S albumingene for sesame and pistachio and for the vicilin precursorgene for macadamia nut. These systems were subjected to a robust intra-laboratory qualitative validation process prior to their application, by DNA extraction and fast real-time PCR, on some real market samples to reproduce a potential allergen contamination along the food chain. The developed system results were specific and robust, with a sensible limit of detection (0.005% for sesame; 0.004% for pistachio; 0.006% for macadamia nut). The performance and the reliability of the target systems were confirmed on commercial food samples. This molecular approach could be used as a screening or as a support tool, in association with the other widespread monitoring techniques (such as ELISA).

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Digital PCR for analysis of peanut and soybean allergens in foods

Cited by 23 publications

References 35 publications

Peanut Allergy: Characteristics and Approaches for Mitigation

Peanut Allergy: Characteristics and Approaches for Mitigation

Fluorescence‐based quantitative platform for ultrasensitive food allergen detection: From immunoassays to DNA sensors

Sesame, Pistachio, and Macadamia Nut: Development and Validation of New Allergenic Systems for Fast Real-Time PCR Application

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