Real Time PCR for the detection and discrimination of cereal contamination in gluten free foods

Real-time PCR methods, using melting curve analysis for product identification, were established for the specific discrimination of wheat, rye, barley and oats in food samples. Specific primers targeting cereal prolamin genes were chosen for the amplification. The lengths of the amplicons varied between 104 and 181 base pairs and the melting point between 81.2 and 85.0 degreesC. The specificity of the wheat PCR was shown for spring and autumn wheat but also for durum wheat, spelt wheat and kamut. The methods were applied for final food products and for detection of toxic cereal contamination in oat samples. The results of the analysis were compared with those obtained with an established enzyme immuno assay for gluten analysis. The PCR methods give a good correlation with the protein assay and are rapid and sensitive. The PCR methods can thus be used as confirmatory methods in food analysis of gluten-free and naturally gluten-free foods.