Daniele De Rossi will be sent home after limping off with a thigh injury in Italy’s 2-2 draw with Bulgaria.
The midfielder had been a risk to feature beforehand, but recovered from a right ankle problem.
He went off at the 75th minute after clutching the back of his right thigh, leaving the Azzurri with 10 men as all the substitutions had been completed.
“De Rossi felt a twinge in the femoral bicep, so obviously he had to stop,” confirmed chief medic Professor Enrico Castellacci.
“Tomorrow we will perform tests and have to decide when he will leave for Rome. Clearly, his Azzurri adventure has finished for now.
“Therefore he will probably undergo the tests in Rome. We can only evaluate the extent of the damage over the next few days.
“It is the same leg that saw him damage his ankle last week, as it was the right thigh.”
Italy continue their campaign by welcoming Malta to Modena’s Stadio Braglia on Tuesday, but De Rossi will not be taking part.
Claudio Marchisio also sustained an injury during the 2-2 World Cup qualifier in Sofia.
“Marchisio crashed against the advertising board and suffered cuts to his hand and knee, but these are things that can heal fairly quickly.”
Think you know your Italian football? Share your knowledge, tips and comments to win cash prizes in OLBG's World Cup tipster competition - £10,000 to be won!