Rossi has been plagued by knee problems and damaged the anterior cruciate ligament in his right leg in January, leaving his availability for the World Cup into doubt.
However, after featuring as a substitute in this months Coppa Italia final and two subsequent Serie A fixtures, the 27-year-old has been selected by national coach Cesare Prandelli.
Antonio Cassano, who last represented Italy in the final of UEFA Euro 2012, is another player to have made the cut.
There are also call-ups for Torinos Ciro Immobile, Serie As top scorer this season, and Christian Maggio of Napoli, who has recovered from surgery on a partially collapsed lung.
Genoa goalkeeper Mattia Perin, Veronas Brazil-born midfielder Romulo and Torino defender Matteo Darmian are the three uncapped players in the party, while Giorgio Chiellini is included despite receiving a three-match domestic ban for elbowing Miralem Pjanic in Juventus weekend victory over Roma.
However, Stephan El Shaarawy, another player hampered by injury in recent months, Alberto Gilardino and Luca Toni - the scorer of 20 league goals this season - have missed out, together with Pablo Daniel Osvaldo and Emanuele Giaccherini.
Parma goalkeeper Antonio Mirante will join Italys 30-man squad for a training camp in Coverciano as injury cover.
Prandellis men play friendlies against the Republic of Ireland and Luxembourg before facing England, Costa Rica and Uruguay in Group D at the World Cup.
Squad in full:
Goalkeepers: Gianluigi Buffon (Juventus), Mattia Perin (Genoa), Salvatore Sirigu (Paris Saint-Germain), Antonio Mirante (Parma - included as injury cover)
Defenders: Ignazio Abate (Milan), Andrea Barzagli (Juventus), Leonardo Bonucci (Juventus), Giorgio Chiellini (Juventus), Matteo Darmian (Torino), Mattia De Sciglio (Milan), Christian Maggio (Napoli), Gabriel Paletta (Parma),Manuel Pasqual (Fiorentina), Andrea Ranocchia (Inter)
Midfielders: Alberto Aquilani (Fiorentina), Antonio Candreva (Lazio), Daniele De Rossi (Roma), Claudio Marchisio (Juventus), Riccardo Montolivo (Milan), Marco Parolo (Parma), Andrea Pirlo (Juventus), Thiago Motta (Paris Saint-Germain), Romulo (Verona), Marco Verratti (Paris Saint-Germain)
Forwards: Mario Balotelli (Milan), Antonio Cassano (Parma), Alessio Cerci (Torino), Mattia Destro (Roma), Ciro Immobile (Torino), Lorenzo Insigne (Napoli), Giuseppe Rossi (Fiorentina)