Chelsea went into the clash one point ahead of the Bianconeri in Group E but were dominated throughout and found no answer to goals in either half from Fabio Quagliarella and Arturo Vidal.
The champions' hopes of any kind of fightback were dashed when second-half substitute Sebastian Giovinco beat the onrushing Petr Cech in the first minute of injury time to secure a 3-0 win.
"It was a great team effort," said assistant Juventus coach Angelo Alessio, whose side now sit second in the group behind Shakhtar Donetsk after the Ukrainians qualified by defeating Nordsjaelland.
"We were strong in defence but the whole team showed up with the will to put in a strong performance. It has given us a lot of confidence but we still feel there's room for improvement."
Barely six months after being crowned kings of Europe, Chelsea are facing the ignominy of dropping out of the competition before the lucrative last 16 knockout phase.
It comes amid a torrid spell for the English giants, who have now leaked 20 goals in their past nine games in all competitions, including two in a 2-1 defeat to West Bromwich Albion on Saturday.
Blues manager Roberto Di Matteo said: "It's a big disappointment. Obviously a difficult evening for us, the team, the club. A lot of reflection."
Chelsea must beat Nordsjaelland in their final match to qualify, but their last 16 destiny now depends on the result of the Shakhtar v Juve match in a fortnight.
Chelsea started with no recognised striker after leaving Fernando Torres on the bench but Juventus went about their task positively and should have had several goals during a dominant first-half.
Early on Cech had to rush to keep out Stephan Lichsteiner's shot at the back post from Claudio Marchisio's short cross.
Chelsea, however, then missed a great chance to heap pressure on the hosts when Eden Hazard forced Gianluigi Buffon to parry his angled snap-shot off the post after Oscar's superb run past the Juventus defence in the ninth minute.
Juventus were a constant threat but good run down the left by Ashley Cole allowed the left back to feed Oscar on the edge of the area.
After a perfect square ball to Ramires his shot from 20 metres went well wide of Buffon's goal.
Di Matteo explained that he fielded no striker in order to have Hazard, Mata and Oscar changing positions to confuse Juve's defence.
"I didn't want to give the centre-backs any points of reference," he added.
"Hazard had the first good chance for us, Oscar had another one and Mata too.
"In tight games the first goal is often decisive."
When Juventus scored, it was in textbook fashion but came with a dose of good luck, Pirlo hitting a speculative dipping shot that Quagliarella managed to deflect from 15 metres out into the Chelsea net.
Chelsea were lucky not to concede a second a minute later, Cole rushing to clear as Lichsteiner's shot bobbled towards the goalline.
Seconds later Buffon rushed out in timely fashion to deny Mata as the Spaniard ran down on goal unhindered.
After the restart, chances by Vucinic and Vidal went begging and the hosts were denied what appeared to be a valid penalty claim when defender Gary Cahill impeded Mirko Vucinic in the area after Quagliarella's lob put the Montenegrin through.
Di Matteo replaced midfielder Cesar with Victor Moses just before the hour but a minute later Chelsea were 2-0 down after Vidal's first-time shot from Kwadwo Asamoah's perfect cutback took a deflection off Ramires to wrong-footed Cech.
Di Matteo sent on Fernando Torres, but it appeared a desperate move which had little effect.
Chelsea had little to offer and when diminutive midfielder Giovinco was released and went on to run unmarked towards goal he slipped the ball past the ball past the onrushing Cech from long range to seal the win in the 91st minute.
Di Matteo added: "Mathematically, it's still possible. When there's hopes you try, we just have to make sure we do our job our end and win the last group game."