The wine sector is one of the most active and profitable in Italy; Italy is the largest wine producer in the world, followed by Spain and France. For this reason, it is subject to extensive legislative discipline. Calabria ranks sixteenth among Italian regions for wine production; however, in recent years, much effort has been made to promote the wine sector in this region. Nine Calabrian wines are designated as PDO (protected designation of origin) and ten wines as PGI (protected geographical indication). Among the PDO wines, Terre di Cosenza and Ciro are the most famous. Fraud prevention is necessary for consumer protection and for product valorization and promotion. Microsatellite marker analysis represents a reliable and effective tool for varietal characterization, allowing the detection of adulteration in the winemaking process. We sampled 20 grapevine cultivars from different areas of Calabria commonly used for production of the PDO wine Terre di Cosenza and 20 cultivars from other Italian regions in order to carry out a molecular characterization. The analysis was performed through six simple sequence repeat (SSR) markers proposed by the International Organization of Vine and Wine (OIV), VVSS2, VVMD5, VVMD7, VVMD27, VrZAG62, and VrZAG79. Some of the cultivars showed the same molecular profile, revealing the presence of synonymies and incorrect name assignment.