Douglas Costa injured his right thigh in Friday's 2-0 win over Costa Rica and will miss Brazil's final World Cup Group E game against Serbia, the team doctor said on Saturday.
The Juventus winger will not travel to Moscow for Wednesday's game which will be decisive for both teams.
Brazil top the group and will qualify with a draw but the five-times world champion could go out if they lose to the Serbs.
"At the end of the game yesterday Douglas Costa complained of a pain in the back of his right thigh," doctor Rodrigo Lasmar told reporters. "Exams showed he has a slight muscle injury, a diagnosis confirmed today by a magnetic resonance scan."
Lasmar said full back Danilo, who missed the Costa Rica match with a thigh problem, will also stay at the team's base in Sochi for more treatment.