Police were called to Ladys Pass, in the Greater Bendigo region, at 1:25am after reports of a single-vehicle crash on McLennan Street.
The 34-year-old rider of the quad bike from Ladys Pass died at the scene.
Investigations into the crash are ongoing, as police are not sure how the incident unfolded.
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Officers will now prepare a report for the coroner.
Anyone who may have witnessed the collision, and anyone with dashcam footage of the area at the time, has been asked to contact the authorities.