Vlahovic, 26, had been out since December but returned to action in mid-March before suffering this latest setback during Monday's 2-0 Serie A win over Genoa.
According to Italian media, Vlahovic will be out for at least three weeks.
"During the warm-up held in the second half of the match against Genoa, Dusan Vlahovic felt an issue in his left calf," Juve said.
"As a result, this morning he underwent diagnostic tests.
"The examinations revealed low-grade damage," they added.
According to Italian media reports Vlahovic will be out for at least three weeks.
Juve are fifth in the Italian table, one point behind Como in the final Champions League spot with seven games of the season to play.
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