Netherlands midfielder Rafael van der Vaart on Wednesday became the third Dutch player to be ruled out of the World Cup through injury.
Hamburg midfielder van der Vaart, who has 109 international caps, hurt his right calf at the Netherlands' training camp in Portugal.
Following medical tests, Dutch coach Louis van Gaal confirmed van der Vaart would miss the finals in Brazil, according to Dutch magazine Voetbal International's website.
Van der Vaart, 31, joins Roma midfielder Kevin Strootman (knee) and Paris Saint-Germain defender Gregory van der Wiel (tendinitis) on the sidelines with injuries.
The Hamburg midfielder had been set to possibly eclipse Frank de Boer's record number of appearances for an outfield player (112) in Brazil.
Van der Vaart featured at the 2010 World Cup in South Africa, when the Netherlands lost to Spain in the final. He also played in the 2006 finals.
His absence means there are now just seven members from the 2010 team left in van Gaal's provisional squad.
Van der Vaart's exclusion likely means Wesley Sneijder will be included in the final 23-man squad despite van Gaal having previously expressed doubts as to the condition of the Galatasaray playmaker.
Sneijder and PSV Eindhoven captain Georginio Wijnaldum are expected to compete for the No 10 role at the tournament.
The Dutch are in Group B at the World Cup along with defending champions Spain, Chile and Australia.