All Seymour, Shepparton and Albury train services will be replaced by road coaches until further notice.
Passengers travelling between Donnybrook and Southern Cross may consider using metropolitan services as an alternative.
The train was travelling from Sydney to Melbourne when several carriages and the engine derailed near the Hume Fwy just before 8 pm on Thursday.
The driver, a 54-year-old ACT man, and the train pilot, a 49-year-old Castlemaine man, died at the scene.
Twelve passengers were transported to hospital with non-life threatening injuries. A report for the coroner will be prepared.
Police will investigate the cause of the incident in conjunction with National Rail Safety Regulator, Australian Transport Safety Bureau and WorkSafe.
As part of the investigation, the ATSB will examine the track infrastructure, the XPT power cars and carriages and map the accident layout.
The ATSB will also obtain and analyse available information and records, including the train data logger, signalling data and maintenance records for the train and track infrastructure.
Investigators will also consider vehicle crashworthiness and human factor aspects and interview involved parties and witnesses.
A final report will be released at the end of the investigation.
Should a critical safety issue be identified during the course of the investigation, the ATSB will immediately notify those affected and seek safety action to address the issue.
Any passengers and witnesses who may have any information, photographs and video which may be relevant to the investigation are asked to phone the ATSB on 1800 992 986 (then press 1).
Anyone who was on the train and hasn’t spoken with emergency services or who purchased a ticket but did not board the train is asked to phone Seymour police station on 5735 0200.
Passengers should check vline.com.au or the V/Line app for the most up-to-date information before they travel.
The latest public transport information and network status is available at ptv.vic.gov.au or in the PTV mobile app.
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