Police and emergency services responded to a fatal collision in Pyalong late this afternoon.
It is believed a vehicle was travelling along the Northern Hwy about 4.45pm when it veered onto the other side of the road and collided head-on with another car.
Police are yet to establish the exact circumstances surrounding the collision.
The male driver and sole occupant of the first car has been airlifted to hospital with serious injuries.
A female driver of the second vehicle, who was also the sole occupant, died at the scene.
The investigation remains ongoing.
Traffic diversions are in place, with the Northern Highway expected to be closed in both directions south of Moores Lane for several hours.
Anyone who witnessed the collision is urged to contact Crime Stoppers on 1800 333 000 or submit a confidential crime report at www.crimestoppersvic.com.au