The Italian Football Federation (FIGC) confirmed that Marchisio has been tested and revealed severe damage to his right knee and is expected to be sidelined for six months.
A statement from the FIGC said: "During the heating (Friday), Marchisio has suffered an injury to his right knee.
"An MRI was performed immediately in Florence and revealed a subtotal lesion of the anterior cruciate ligament.
"After informing the medical staff of Juventus, the player will return to his club."
His injury is a blow for Italy and Juventus, who are still alive in three races this season.
Juve are 14 points clear of Roma at the top of Serie A with 10 games remaining.
Massimiliano Allegri's team is in the quarterfinals of the Champions League and the Italian Cup semi-finals.
The Turin giants take on the high-flying Ligue 1 Monaco continues club competition in Europe.
The 29-year-old scored two goals and set up more than five in 27 league games for the leaders of Serie A.
Italy take on Bulgaria in Sofia in Euro 2016 qualifying match on Saturday before hosting England in an international friendly in Turin three days later.
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