Identification and quantification of celery allergens using fiber optic surface plasmon resonance PCR

Daems, Devin; Peeters, Bernd; Delport, Filip; Remans, Tony; Lammertyn, Jeroen; Spasic, Dragana

doi:10.3390/S17081754

Title

Identification and quantification of celery allergens using fiber optic surface plasmon resonance PCR

Author

Daems, Devin

Peeters, Bernd

Delport, Filip

Remans, Tony

Lammertyn, Jeroen

Spasic, Dragana

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.

Language

English

Source (journal)

Sensors. - -

Publication

2017

ISSN

1424-8220

DOI

10.3390/S17081754

Volume/pages

17 :8 (2017) , 10 p.