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Barcelona have confirmed Ivan Rakitic has been diagnosed with a hamstring injury.
Rakitic was substituted midway through the second half of Croatia's 3-2 Nations League win against Spain, in which Tin Jedvaj scored a late winner.
Coach Zlatko Dalic subsequently confirmed Rakitic would not travel with Croatia to London for Sunday's decisive clash with England, reports Goal.
And Barcelona announced on Saturday scans have found Rakitic has a hamstring injury in his right leg, with no timescale given for his recovery.
Rakitic was already unavailable for Barca's next game, away to LaLiga title rivals Atletico Madrid on November 24, due to suspension.
The midfielder was sent off in Barca's last match before the international break, a 4-3 loss to Real Betis that was their first home defeat under Ernesto Valverde.
Dalic named Nikola Vlasic, Mateo Kovacic, Josip Brekalo and Milan Badelj as players who could fill in for the injured Rakitic at Wembley.
Croatia will qualify for the finals of the inaugural Nations League if they beat England in a repeat of the World Cup semi-final, but will be relegated to League B if they lose.