Bendigo Police have confirmed that a 28-year-old Epsom man and a 35-year-old Ballarat woman lost their lives in a multi-vehicle collision in Harcourt North on Friday night.
The death's bring the state's road toll to 72, compared to 77 this time last year.
Police say the cause of the collision remains under investigation.
There are reports a vehicle was seen driving southbound in the northbound lanes shortly before the crash, although Bendigo Police would not confirm this.
An Ambulance Victoria spokesperson said it received a call at 9.11pm on Friday night to attend a multi vehicle collision in Harcourt North.
The spokesperson says three people were assessed at the scene, with two people taken to Bendigo Hospital.
A man, believed to be in his 50s, suffered a shoulder injury and was transferred to hospital in a stable condition.
A woman, believed to be in her 20s, suffered a chest injury and was taken to Bendigo Hospital in a stable condition.
Another man, believed to be in his 60s, was injured but did not need to go to hospital.
Police have not released the names of the dead persons.
The exact cause of the collision is yet to be determined.
Police are appealing for any witnesses or anyone with dash cam footage to contact Crime Stoppers on 1800 333 000 or submit a confidential report at crimestoppersvic.com.au