The seamer complained about stiffness in his hamstring during a first-class fixture for New South Wales against Victoria but it was initially cleared of anything serious.
A second scan however confirmed the an injury and Hazlewood will not travel to Perth with Queensland seamer Michael Neser added to the squad.
A statement by Cricket Australia said: "Initial scans Wednesday were clear of muscle strain, however follow-up imaging today has confirmed the injury.
"Early imaging can occasionally underestimate low-grade muscle injuries."
Australia captain Pat Cummins had already been ruled out of at least the opening Test as he recovers from a back injury.
With two senior pacemen ruled out, Mitchell Starc and Scott Boland now become important for the hosts with Brendan Doggett set for a Test debut.