Legislation requires labelling of foods containing allergenic ingredients. We have developed a robust ten-plex quantitative and sensitive ligation-dependent probe amplification method, the allergen-MLPA method, for specific detection of eight allergens: sesame, soy, hazelnut, peanut, lupine, gluten, mustard and celery. Ligated probes were amplified by PCR and amplicons were detected using capillary electrophoresis. Quantitative results were obtained by comparing signals with an internal positive control. The limit of detection varied from approx. 5 to 400 gene copies depending on the allergen. The method was tested using different foods spiked with mustard, celery, soy or lupine flour in the 1-0.001 % range. Depending on the allergen, sensitivities were similar or better than those obtained with qPCR. The allergen-MLPA method is modular and can be adapted by adding probe-pairs for other allergens. The DNA based allergen-MLPA method will constitute a complementary method to the traditional protein based methods.

Determination of allergens in food by multiplex, quantitative ligation-dependent probe amplification

243rd National Meeting (2012)

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