Dumfries was injured playing for Inter Milan in Sunday’s Serie A triumph at Atalanta and has been replaced by 22-year-old Ajax defender Youri Baas who received a first national team call-up on his birthday.
Dutch coach Ronald Koeman also added Mats Wieffer of Brighton & Hove Albion as cover for key midfielder De Jong, who missed Barcelona’s win at Atletico Madrid on Sunday because of illness.
De Jong joined up with the Dutch squad on Monday but did not train.
The Netherlands host Spain in the first leg of their quarter-final tie in Rotterdam on Thursday with the return in Valencia on Sunday.