Abstract
Accurate identification and quantification of allergens is key in healthcare, biotechnology and food quality and safety. Celery (Apium graveolens) is one of the most important elicitors of food allergic reactions in Europe. Currently, the golden standards to identify, quantify and discriminate celery in a biological sample are immunoassays and two-step molecular detection assays in which quantitative PCR (qPCR) is followed by a high-resolution melting analysis (HRM). In order to provide a DNA-based, rapid and simple detection method suitable for one-step quantification, a fiber optic PCR melting assay (FO-PCR-MA) was developed to determine different concentrations of celery DNA (1 pM–0.1 fM). The presented method is based on the hybridization and melting of DNA-coated gold nanoparticles to the FO sensor surface in the presence of the target gene (mannitol dehydrogenase, Mtd). The concept was not only able to reveal the presence of celery DNA, but also allowed for the cycle-to-cycle quantification of the target sequence through melting analysis. Furthermore, the developed bioassay was benchmarked against qPCR followed by HRM, showing excellent agreement (R2 = 0.96). In conclusion, this innovative and sensitive diagnostic test could further improve food quality control and thus have a large impact on allergen induced healthcare problems.
Highlights
Celery (Apium graveolens) is one of the most common foods causing allergic reactions in European countries, in France, Switzerland and Germany
Celery was added to the list of fourteen groups of allergenic species with strict labelling legislation issued by the EU Commission
Api g 1 codes for the major allergenic protein of celery, mannitol dehydrogenase (Mtd) is more widely recommended for use in these assays due to the lower risk of cross-reactivity with sequences of other related species [10,13]
Summary
Celery (Apium graveolens) is one of the most common foods causing allergic reactions in European countries, in France, Switzerland and Germany. In France, 30% of anaphylactic reactions are caused by the ingestion of celery [1,2,3]. Celery is consumed cooked or processed, compared to raw celery, food processing only lowers the allergenicity, but does not destroy the allergenic properties completely [2]. Celery was added to the list of fourteen groups of allergenic species with strict labelling legislation issued by the EU Commission (Directive 2003/89/EC and Directive 1169/2011) in order to ensure that consumers are adequately informed on the presence of allergenic ingredients in food [4,5].
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