Needing a win to leapfrog back ahead of Napoli in the race for second (and automatic Champions League qualification) and in the process delay the Bianconeri's title celebrations, the Giallorossi did just that in a thrilling game - despite going behind early on to Mario Lemina's close-range finish.
Daniele De Rossi, captain on the night, scored just moments later to rescure the momentum for his team - before second half goals from Stephan El Shaarawy and Radja Nainggolan drove the hosts to a memorable and deserved win.
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Only time will tell just how crucial the result will prove to be - it will take something of a miracle for Juventus to slip up in their pursuit of the title, but second remains a keenly-fought contest. Yet nevertheless it was a result to savour in its own right, with the Giallorossi giving a glimpse of their true capabilities.
Without the services of top goalscorer Edin Dzeko, the hosts initially struggled to create real danger in the final third. But De Rossi's equaliser - bundling home after Gianluigi Buffon had saved Kostas Manolas's header - seemed to give the side new confidence, and in the second half they came out with real intent.
It was El Shaarawy who changed the game, as he cut inside and hit a low, curling shot that somehow crept inside the far post. Ten minutes later it was Nainggolan finding the same side of the goal - the midfielder producing a trademark rocket to beat Buffon after being played in by Mohamed Salah.
The result means Roma are now four points behind Juventus in the Serie A standings with two games remaining - while they are back a point ahead of Napoli with six points still up for grabs.
Coach Spalletti made two changes from the side that beat AC Milan so emphatically - one of them enforced, with the injured Edin Dzeko replaced by Stephan El Shaarawy.
The other change came in defence, where Antonio Rudiger returned to the starting XI after suspension - slotting back in at right-back as Emerson shifted back across to his more familiar left-back role.
7' - The hosts with the early pressure. From a counter, Asamoah smacks a drive against the post.
15' - Combative start from both sides, with few clear-cut openings. Higuaín fires over from distance.
18' - Chance for Roma - De Rossi meets Paredes' free-kick but the header is deflected narrowly wide.
21' - Goal for Juventus. Higuaín squares for Lemina who slides home. 0-1.
25' - GOAL! ROMA LEVEL! De Rossi makes it three goals in three games, as he bundles home from close range after Manolas's header is parried away! 1-1!
35' - Solid save from Szczesny to palm away Higuaín's drive from 25 yards out.
42' - Good chance for Roma. Radja finds Salah with a clipped ball into the box but the winger's header is held by Buffon
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56' - GOAL! ROMA LEAD! YES! Brilliant from El Shaarawy, as he latches onto Radja's pass, cuts inside and curls a low shot inside the far corner! Roma lead! 2-1!
66' - GOAL! AGAIN! An emphatic finish from Radja Nainggolan! Salah finds the midfielder inside the box and he absolutely lashes home! 3-1!
84' - Some slightly desperate defending from Roma, but they get the ball away eventually...
90' - Five minutes of additional time. Emerson flashes over from distance after a dangerous counter.
90' - Good save down low from Szczesny to deny Higuaín!
See more game statistics in our dedicated match centre.
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“For some time, there’s been talk about things other than our season. Now it’s absolutely key to win our next two games. Previously we only spoke about the team and our matches. We felt support today. It’s not that we win because of them or have lost at times because of them, but seeing a full stadium like this makes you work harder.”
- Daniele De Rossi
“We said this ahead of the game: it’s not just the win that counts. We wanted to show who we are, to show that we have the determination to change things, and that we can keep on working, believing in our ability to reach our objective sooner or later."Winning this game is testament to what these guys are worth."
- Luciano Spalletti
The Giallorossi have two games remaining in this season - with their final away game coming next Saturday, as they travel to face Chievo Verona on Saturday evening.
The final game of the season takes place the following weekend - exact kick-off date and time is still to be confirmed by the league - against Genoa at the Stadio Olimpico.
Goalscorers: De Rossi (25'), El Shaarawy (56'), Nainggolan (65'); Lemina (21')
Roma: Szczesny; Rudiger, Fazio, Manolas, Emerson; De Rossi, Paredes; Perotti (Grenier), Nainggolan (Jesus), Salah (Totti); El Shaarawy.
Juventus: Buffon; Lichtsteiner (Alves), Benatia, Bonucci, Asamoah; Lemina, Pjanic, Sturaro (Dybala); Cuadrado (Marchisio), Higuain, Mandzukic.
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