For the routine detection of allergens in foods, PCR and/or ELISA methods are employed. To assess the suitability of these methods, proficiency tests (PTs) could be used as a valuable instrument. It is a common practice to evaluate the results with respect to the experimentally obtained robust mean without considering the actual allergen content. In the present study, an overview is given of the results of allergen PTs for the determination of soy and gluten conducted by Dienstleistung Lebensmittel Analytik GbR (DLA). A total of 16 PTs were evaluated with respect to the comparison of PCR and ELISA performances and a new focus on the actually spiked values. The analytes were added in the ranges of 7.8-6264 mg/kg (gluten) and 184-5500 mg/kg (soy protein) in differently composed matrices such as pastry, infant food, and sausage meat. The evaluation of the PTs showed a widely reliable qualitative detection of both allergens by PCR methods. ELISA performances differed for soy and gluten. Although a high number of false-negative results occurred for the detection of soy, the qualitative detection of gluten was appropriate. Quantitative results showed obvious test kit-specific differences for the ELISA methods, but the limits of quantification were suitable for gluten determination. Both ELISA and PCR methods demonstrated their valuable contribution in food allergen determination.

Background: Little is known about breast milk as a vehicle for tolerance development or sensitization to peanuts very early in life. Thus, well-characterized and highly sensitive detection systems for the reliable determination of peanut allergens in breast milk are mandatory. Methods: For the quantification of the marker allergens Ara h 2 and Ara h 6 in the low nanogram per milliliter range in breast milk samples of a German cohort, sensitive and highly specific sandwich ELISAs were optimized and validated. Results: The Ara h 2 ELISA revealed a limit of detection (LOD) of 1.3 ng Ara h 2/mL and a quantification range of 2.3-250 ng/mL, the Ara h 6 ELISA showed an LOD of 0.7 ng/mL and a working range of 1.1-14.4 ng/mL. The assays showed no relevant cross-reactivity against other potentially cross-reactive legume, seed, and tree nut extracts (<0.01%, except for Ara h 1 in the Ara h 2 ELISA <0.1%). Ara h 2 was detectable in breast milk samples from 14/40 (35%) of the participants in concentrations from 2.3 to 184 ng/mL, Ara h 6 appeared in 9/40 (22.5%) of the lactating mothers between 1.1 and 9.7 ng/mL, and 1 highly positive sample with 79 ng/mL. Both allergens appeared at the same time points, but Ara h 6 in lower concentrations than Ara h 2. Conclusions: Sensitive and specific diagnostic tools for the determination of Ara h 2 and Ara h 6 in human breast milk were established. The kinetics of secreted Ara h 2 and Ara h 6 seem to be similar but with a difference in concentration. Follow-up investigations on their tolerogenic or sensitizing properties in breast milk become now accessible.