Validation and application of a quantitative real-time PCR assay to detect common wheat adulteration of durum wheat for pasta production

- Gliadin gene is not an appropriate common wheat (T. aestivum) specific target.
- Grain quantitative kit guarantees target detection, specificity, and sensitivity.
- The validated method is an effective system to quantify soft wheat in durum wheat.
- Commercial wheats and flours were analyzed to detect wheat adulteration.

Pasta is the Italian product par excellence and it is now popular worldwide. Pasta of a superior quality is made with pure durum wheat. In Italy, addition of Triticum aestivum (common wheat) during manufacturing is not allowed and, without adequate labeling, its presence is considered an adulteration. PCR-related techniques can be employed for the detection of common wheat contaminations. In this work, we demonstrated that a previously published method for the detection of T. aestivum, based on the gliadin gene, is inadequate. Moreover, a new molecular method, based on DNA extraction from semolina and real-time PCR determination of T. aestivum in Triticum spp., was validated. This multiplex real-time PCR, based on the dual-labeled probe strategy, guarantees target detection specificity and sensitivity in a short period of time. Moreover, the molecular analysis of common wheat contamination in commercial wheat and flours is described for the first time.