Collaborative project: Development of innovative rapid test and screening methods for on-site detection of food-borne allergens in product development and control – Subproject 1 | FISA - Forschungsinformationssystem Agrar und Ernährung

Collaborative project: Development of innovative rapid test and screening methods for on-site detection of food-borne allergens in product development and control – Subproject 1

Project

Project code: 2816400508, BfR-LMS-08-1337-196, PGI-06.01-28-1-64.005-08
Contract period: 01.09.2009 - 28.02.2013
Budget: 303,904 Euro
Purpose of research: Experimental development

The aim of the project is the development of innovative, molecular biology based methods for the detection of allergens which are subject to declaration in food. Apart from immunological rapid tests, which can be performed on-site without advanced equipment or specially trained staff, for the first time a new approach building up on a rapid DNA test format shall be applied. A further goal towards method improvement is to establish product group specific 'screening' tools, which are also suited to develop official standardized protocols according e.g. to the German foodstuffs act (§64, LFGB) and which can be employed to check for legally required correct labelling. With respect to the introduction of a threshold currently under discussion, the rapid tests shall be characterized by a sufficient sensitivity to enable the detection of allergens in trace amounts in a relevant range (ppm) and will be characterised by defined detection, hence exclusion limits.

Based on the principle of a „Ready-to-Use“approach, a qualitative real-time PCR (polymerase chain reaction) method was developed for the rapid detection of allergens in food. All substances which are needed to perform the method (primer/probes) so as - if appropriate - additional MgCl2 - are pre-spotted together with the bioprotective substance trehalose in a mass relation of 20:1 (nucleic acid/trehalose) onto the cavities of a PCR microtiter plate. The user only has to add master mix and DNA extract. The pre-prepared PCR plates can be stored at room temperature for many months. It was demonstrated that even under an extreme temperature of 50o C for three month the plates did not show significant loss in efficiency of the PCR-systems. It was shown with the example of confectionary and bakery goods or meat products, that up to eight different allergenic species could be detected simultaneously by using the assay. Moreover, in the frame of the project new PCR systems have been developed for the detection of difficult product groups like molluscs and crustaceous animals. The flexible, validated screening procedure can be applied in routine analysis by the official food control as well as in quality survey during processing. The very good reproducibility of PCR results derived from different runs will be proofed with external laboratories in a ring trial.

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