De Jong hobbled out of Sunday’s dramatic 2-1 win over Mexico in the round of 16 after just nine minutes due to a groin injury.
And an MRI scan has confirmed a tear that is set to sideline the AC Milan man for two to four weeks.
In a best case scenario, De Jong retains a slim chance of regaining fitness in time for the World Cup final on July 13 if the Dutch are able to make it that far.
The 29-year-old will stay in Brazil with Louis van Gaal's squad as he begins his recovery.
A statement on the official website of the Dutch Football Association (KNVB) read: "MRI examination at the hospital confirmed that Nigel de Jong is struggling with a tear in his groin muscle. The 75-caps international is therefore presumably off for two to four weeks.
"De Jong, who last Sunday in the eighth-finals against Mexico (was) injured after nine minutes remains with the team in Brazil to work there (on) his rehabilitation."
Former Hamburg and Manchester City player De Jong impressed in playing every minute of Netherland's Group B wins over Spain, Australia and Chile.
He was replaced by Bruno Martins Indi in Fortaleza at the weekend and the Feyenoord man appears well-placed to start Saturday's quarter-final versus Costa Rica in Salvador.
