Salvatore Sirigu made a crucial error that allowed Florin Andone to score a late equaliser as Romania held Italy to a 2-2 draw in Bologna.
The Paris Saint-Germain goalkeeper had only been on the field for 20 minutes, as a substitute for veteran Gianluigi Buffon, when he parried the ball into the path of Andone who levelled the match in the 89th minute.
Bogdan Stancu's first-half goal for Romania was cancelled out by a Claudio Marchisio penalty, before Manolo Gabbiadini scored his first international goal and Italy looked to have completed an impressive turnaround.
But Antonio Conte's final substitution of the match proved to be decisive, when Sirigu failed to hold Vlad Chiriches' header and Andone struck from point-blank range to send the Romanian supporters home happy.
The result was hard-earned by a Romania side that seemed determined to defy their position one place below Italy in FIFA's world rankings with a confident performance, and Conte's men were left with a defeat and a draw to show for their two November friendlies.
Italy had Buffon to thank for keeping the scores level when the veteran goalkeeper was at full stretch early on to save from Gabriel Torje's sweetly struck drive.
But Buffon could do nothing to prevent Stancu opening the scoring moments later, after a mix up between Matteo Darmian and Andrea Barzagli allowed the Genclerbirligi forward to dart into the box and side-foot a cool finish into the net.
The home team showed no lack of urgency as they looked to bounce straight back into the contest, and Alessandro Florenzi delivered a cross that flashed across the face of goal as Eder slid in just too late to convert it.
Italy's best chance of the half went begging when Eder made a great run to the byline and sent over an inch-perfect cross, but Stephan El Shaarawy got the ball tangled up in his feet and failed to test Ciprian Tatarusanu with a shot.
Romania's lead looked precarious just before half-time, as Marchisio rifled a shot just over the crossbar and Eder had a strong penalty claim turned away, but the hosts went in at the break with nothing to show for their efforts.
Within 10 minutes of the restart, Italy were given a lifeline when Dragos Grigore felled Eder in the penalty area and Marchisio struck a near-perfect spot-kick, sending the ball into the top corner off the crossbar.
Conte then brought Eder off as part of a triple substitution that saw Gabbiadini step up off the bench, and within six minutes of his introduction the Napoli winger was presented with the easiest of chances and he headed home unchallenged from five yards out.
But just as Italy looked to have wrapped up their fifth win in six games, Chiriches' header from Lucian Sanmartean's free-kick was parried by Sirigu to the feet of the grateful substitute Andone, who made no mistake from inside the six-yard box.