"For every decision we make in our return to play, the well-being of our players, staff, officials, and all participants is our top priority," MLS commissioner Don Garber said.
The expansion side is now the second team to be pulled from the competition after FC Dallas was sent home on Monday. Dallas announced that 10 players and a member of the technical staff tested positive following the club's arrival in Orlando for the competition, which initially had 26 teams involved.
Nashville's Group A game against the Chicago Fire was postponed following the confirmation of five cases. Reports then surfaced Wednesday evening regarding four more players returning positive results following an additional round of testing.
With Dallas and Nashville removed from the competition, the format has been reconfigured into six groups of four teams each. Chicago will now join Group B alongside Seattle, Vancouver, and San Jose.
The tournament kicked off Wednesday and was originally slated to feature 54 matches behind closed doors over a month at the Walt Disney World Resort near Orlando.
Several players have expressed concerns about resuming the 2020 MLS campaign amid a pandemic in an area of the United States that's been hit particularly hard. LAFC star and reigning MLS MVP Carlos Vela opted to stay home to be with his family, and teammate Bradley Wright-Phillips called the tournament a "little bit stupid."