A reliable assay for the detection of soft wheat adulteration in Italian pasta is based on the use of new DNA molecular markers capable of discriminating between Triticum aestivum and Triticum durum

Italian pasta must be prepared using exclusively durum wheat. According to current Italian rules, only a maximum of 3% Triticum aestivum is allowed to account for cross-contamination that may occur during the agricultural process. Efficient methods for the detection of accidental or intentional contamination of common wheat to durum wheat products are therefore required. This article describes a novel approach for the detection and quantification of soft wheat adulteration in whole grain durum flours and dried pasta. The assay relies on the presence of intron-specific DNA length polymorphisms in the plant β-tubulin gene family, which can be highlighted through the PCR-based TBP (Tubulin-Based Polymorphism) method. In wheat, the TBP method produces species-specific amplification products, which can be either directly used as new DNA molecular markers capable of discriminating between T. aestivum and Triticum durum or analyzed at the sequence level for the design of species-specific probes. The latter approach allowed the development of new sequence-specific targets that can be exploited in RT-PCR assays for a rapid and accurate quantification of soft wheat adulteration in durum wheat pasta.