The KNVB released a brief statement on their official website announcing the injury, and that Van der Vaart would not be included on the final 23-man roster.
"The Holland international has a calf strain in his right leg. Coach Louis van Gaal has therefore decided not to include him in the final selection."
The 31-year-old was hoping to make his third straight World Cup team, having played in the tournament in both 2006 and 2010. He currently plays for Hamburg of the Bundesliga.
The Dutch were already missing Kevin Strootman and Gregory van der Wiel, both of whom were already ruled with injuries. They begin Group B play on June 13th in rematch of the 2010 World Cup final against Spain.