Midfielder Tonny Vilhena left the Dutch international camp on Tuesday and will miss the game against Spain after testing positive for coronavirus.
The player "will return home immediately and await further test results", announced the Dutch FA.
Vilhena, 25, is a regular starter for Russian club Krasnodar but has struggled for minutes for the national team. His last start came in a friendly against Italy in June 2018.
Holland now have a squad of 23 players for their friendly against Spain, followed by Nations League clashes with Bosnia and Herzegovina and Poland.
Vilhena is the fifth player to leave the camp with an injury or illness after Berwijn, Cillessen, Bijlow, and Ihattaren.