The Serie A side will come up against the Dutch champions on Tuesday in Group H and will be without the 20-year-old.
El Shaarawy looked set to feature after failing to play a part in their opening 2-0 victory over Celtic on September 18 with a thigh injury, but suffered another blow in training.
He will now not only miss out on their fixture in Amsterdam, but could also be a doubt for their clash with Barcelona later in the month and the news is frustrating for Allegri.
"Stephan took a knock in training and felt some pain," he said.
"As a precaution we sent him for a scan, and there it was revealed that he has suffered a microfracture which will keep him out for 20-30 days.
"It's a shame because he was back in form after a muscle problem and we were counting on him for this match.
"We'll have (Mario) Balotelli back, but I think his recent suspension has shown him that he has to change his attitude on the pitch.
"In Europe there are no easy games. Ajax play some good football and have a great tradition. We'll need to be very careful."
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