Federico Chiesa, one of the stars of Italy's successful Euro 2020 campaign, is unlikely to be available as he bids to avoid a humiliating failure to reach this year's Qatar World Cup.
Chiesa suffered an anterior cruciate ligament injury to his left knee while playing for Juventus against Roma in Serie A on Sunday.
He will undergo surgery due to the possibility of Italy crashing out of the play-off process in March after a poor qualifying group stage.
Juventus said he felt a "blunt sprain" in his knee, and was tested on Monday.
Italy will host North Macedonia in their four-team play-off category on 24 March. If they win they are likely to beat Portugal to reach Qatar.
Chiesa will miss his club's two-leg last-16 Champions League matches against Villarreal starting next month.
He will also be absent for several Serie A matches. Juventus, which are in fifth place, are 11 points behind leaders Inter Milan and three points from the Champions League qualifying places.
Chiesa Juventus beat Roma 4-3 despite beating coach Jose Mourinho, prompting coach Jose Mourinho to blame his players' lack of mental strength.
At the end of the day, when you're in the dirt, you get back on your feet and find your character," Mourinho told DAZN. "But there are people in this changing room who are a little bit too cool, A little too much. Weak.
"The talk of playing in the comfort zone, playing six to seven out of 10, is too easy for some celebrities. We need to get out of this mindset."