Goalkeeper Gianluigi Buffon, defensive trio Andrea Barzagli, Giorgio Chiellini (pictured with Buffon) and Leonardo Bonucci and midfielders Andrea Pirlo and Claudio Marchisio are all from the Turin club.
The Italians have already qualified from Group C with two matches to spare and Prandelli has decided to field an adventurous 4-4-2 formation.
"The match with Serbia should be a good barometer of our credentials because they are technically one of the best teams in Europe," Prandelli told a news conference on Thursday.
"We will be fully motivated because there is strong competition in our squad for spots in the starting line-up and this game is the start of our preparations for the finals.
"Giuseppe Rossi has shaken off his injury, he went the distance in a very intense training session, so he will start up front alongside Antonio Cassano," added Prandelli.
Serbia coach Vladimir Petrovic is likely to stick to a more conservative 4-2-3-1 formation.
His team will seal a play-off berth with a win although a draw will also be enough if Estonia fail to beat Northern Ireland in Belfast.
"The Italians are lethal on the break and we have to be smart and patient in order to avoid falling behind because that would make life very difficult for us," Petrovic said.
"We will be in touch with what's going on in Belfast because the outcome of that match is very important for us but the priority is to stay completely focused on our own task."
Italy have 22 points from eight games followed by Serbia on 14 from eight.
Estonia have 13 from nine, Slovenia have 11 from nine and Northern Ireland have nine points from eight matches.
The winners of Europe's nine qualifying groups and the best second-placed team gain automatic berths in the finals alongside joint hosts Poland and Ukraine while the other eight runners-up enter two-legged play-offs for the remaining four places.
Team: Gianluigi Buffon; Christian Maggio, Giorgio Chiellini, Andrea Barzagli, Leonardo Bonucci; Riccardo Montolivo, Daniele De Rossi, Andrea Pirlo, Claudio Marchisio; Giuseppe Rossi, Antonio Cassano.
