Messi was in action for Inter Miami in their 2-1 win over Atlanta United at the Mercedes-Benz Stadium.
It was Atlanta's big-money January recruit Emmanuel Latte Lath who opened the scoring in the 11th minute.
Latte Lath, who joined Atlanta for £17million from Championship side Middlesbrough, made an intelligent move towards the back post and rose highest to nod home Brooks Lennon's floated cross.
But Atlanta's lead lasted just nine minutes as Messi rolled back the years to produce a moment of magic.
Messi raced in to dispossess Bartosz Slisz on the edge of their 18-yard box.
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The Argentinian superstar then sent Derrick Williams sliding the wrong way as he chopped the ball onto his right foot and left him with only Guzan to beat.
The former Aston Villa goalkeeper got as far as his six-yard box and went down to the ground in an attempt to smother Messi's shot.
Unfortunately for Guzan, Messi left him red-faced as the eight-time Ballon d'Or winner cheekily chipped the Atlanta gloveman for the equaliser.
It was Messi's fifth goal against Guzan in seven games since the former moved to Inter Miami in July 2023.
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Unfortunately for Guzan, his day was about to end on a more sour note.
Former Barcelona defender Jordi Alba sent in a cross from the left towards the back post where Haitian international Fafa Picault was waiting.
Picault did not get the most powerful contact on the header but managed to place it perfectly across goal.
Guzan desperately scrambled across his line to keep Picault's header out but could not get there in time, as the 34-year-old raced over to celebrate with Miami's bench in the 89th minute.
The American goalkeeper cut a devastated figure as he fell on his back while Atlanta's players stood around with hands on their heads.
Despite Atlanta's efforts they could not find a late equaliser, as Javier Mascherano's Miami picked up their third win of the season.
Although Miami got the three points against Atlanta, Guzan and his teammates can take comfort in the knowledge they knocked out their rivals in last season's MLS Cup playoffs.
Miami and Atlanta traded 2-1 victories for the first two games to set up a mouthwatering eliminator match and one where Messi and Guzan almost came to blows.
Messi popped up in the 65th minute to level the scores at 2-2 and barged into Guzan to get the ball back.
Guzan attempted to intimidate Messi and stand over him but was clattered into by Leonardo Campana.
Unfortunately for the American, he got stuck in his own net when trying to get out as Luis Suarez ran over and pointed in his face.
Yet Guzan had the last laugh as Slisz scored what proved to be the winner in the 76th minute to secure a 3-2 win and send Miami packing from the first round of the play-offs.