American coach Maurice Pochettino made the announcement during an interview with Fox about 90 minutes before the kickoff at Lumen Field in Seattle.
The move is a setback for a U.S. team riding high after defeating Paraguay 4-1 in its first game one week ago.
Pulisic left the game at halftime. Pochettino said the 27-year-old attacker was kicked in the back of his left calf during a training session ahead of the game and felt tightness during the match.
Pulisic trained on his own in the lead-up to the game against Australia.
Pulisic has 33 goals in 87 international appearances.