De Rossi completed 90 minutes of his country's shock 1-0 defeat to Group D surprise package Costa Rica but sustained bruising to his calf.
And the Roma midfielder is set to be restricted to a watching brief in Natal on Tuesday, for a match that will decide who takes the second qualifying spot behind the Central American nation.
"It is very likely De Rossi won’t be able to play against Uruguay," professor Enrico Castellacci told a press conference, before explaining De Rossi's participation in the rest of the tournament - should Italy progress - is not in jeopardy.
"There are no lesions to the soleus muscle. It simply is an injury that requires a few days to recover from.
"He felt this pain during the second half and it intensified towards the end of the game. The tests showed bruising on the calf muscle that will very likely stop him from playing against Uruguay."
There was better news from Castellacci on Milan full-back Mattia De Sciglio, who will come into the coach Cesare Prandelli's plans after shaking off the thigh problem that ruled him out of the opening two games.
Elsewhere, midfielder Claudio Marchisio and defender Andrea Barzagli are also expected to be available despite knee complaints. Barzagli nursed a bout of tendinitis through the Costa Rica match.
Castellacci added: "De Sciglio has recovered well from the bicep problem, so he is at the disposal of the coach.
"Claudio Marchisio had a small problem in his knee, but nothing to worry about.
"Andrea Barzagli is being monitored moment by moment. He wasn't in bad shape today. If his condition remains like this, then he can play."
