The Ghanaian international was brought on for Lemar in the 71st minus.
Atletico led through Alvaro Morata and Antoine Griezmann, but Valencia replied through Kevin Gameiro and a Dani Parejo penalty, awarded by VAR for handball.
Just as the game looked set for a draw, Partey slipped a wonderful ball to Angel Correa on the edge of the box who found the right side of the net with a great finish.
Barca will be champions if they beat Levante at home on Saturday and could seal the title before then if Atletico lose at home to Real Valladolid earlier in the day.
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