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.
Brendan Doggett has expressed confidence in his own game as he prepares for a potential Test debut for Australia in the Ashes opener against England in Perth on 21 November.
Australia seamer Josh Hazlewood has been cleared to continue preparing for the first Ashes test after he and Sean Abbott went for scans on Wednesday.
Jake Weatherald has received his maiden international call-up as Australia named a 15-man squad for the first Ashes Test against England, starting on 21 November in Perth.
Josh Hazlewood starred with the ball as Australia beat India by four wickets at the MCG to go 1-0 ahead in the T20I series.
