The Paris Saint-Germain full-back limped out of Tuesday's Coupe de France final with a knee injury.
Original assessments suggested that though the injury would end Alves' domestic season he would be ready for the World Cup.
"Following Daniel Alves' right knee injury, which occurred yesterday during the French Cup final, the tests performed today showed a high detachment of the anterior cruciate ligament with posterolateral sprain," a spokesperson for the player said on Wednesday.
And the Brazilian Football Confederation (CBF) confirmed on Friday that following further tests by the Brazil national team's doctor, Rodrigo Lasmar, it has been determined the former Barcelona star will not recover in time to represent the Selecao this summer.
"Close examinations confirmed the initial diagnosis of an anterior cruciate ligament injury, requiring surgical treatment," a statement read.
"Considering the alternative recovery options within the timeframe, it confirmed the impossibility of calling Daniel Alves up for the period of preparation, friendlies and the World Cup."
Alves has been a key figure for Brazil for over a decade and was set to start for Tite's side as they looked to secure a sixth world title.
He becomes the second Brazil defender to suffer an serious injury in the build-up to the tournament, with Filipe Luis suffering a broken leg while playing for Atletico Madrid back in March.