The Labor government trails the coalition by 10 points on a two-party preferred basis, according to a Newspoll conducted in October and published in The Australian newspaper on Monday.
The result represents a 10 per cent swing against Labor since the 2022 election and a three-point slide since July, when Premier Ben Carroll took the reins from predecessor Jacinta Allan.
The sobering poll landed on day two of Labor's official ground campaign after Mr Carroll took the red campaign bus for its first spin on Sunday.
On Monday, Health Minister Ingrid Stitt conceded Labor faced an uphill battle in its quest to secure a historic fourth straight term in government.
"We know this is going to be a tough election," she told reporters in Melbourne.
Hours prior, federal Labor counterpart Mark Butler told ABC TV the state government's 12-year legacy, including the harsh lockdowns of the COVID-19 years, made securing re-election "a tough one".
But Mr Carroll remained optimistic, insisting the only poll that mattered was the state election, due on November 28.
"The polls ... had Jeff Kennett comfortably ahead in 1999 and had the Essendon Footy Club comfortably ahead to win the AFL Grand Final," he said.
"There's another poll called the door-knocking poll."
Speaking from across town, Opposition Leader Jess Wilson said she felt confident 35 days from the state election but would not rest on her laurels.
"I'm encouraged about the opportunity ahead (but) my team takes nothing for granted," she told reporters.
Among the 1017 Victorian voters surveyed online for the latest Newspoll, Labor's primary support fell to 24 per cent, against the coalition's 33 per cent share.
That represented a lift of two per cent for the Liberal-Nationals, while One Nation's vote edged up one point to 20 per cent.
The result also solidified the opposition leader's box-seat position to become the next premier.
Nearly half of voters indicated Ms Wilson was their preferred leader, while Mr Carroll polled 34 per cent.
It also makes it more likely the coalition would be able to secure majority government without relying on the help of One Nation MPs.
Despite the changed outlook, Mr Carroll repeated his claim Ms Wilson would need to strike a deal with the minor party.
"We know that Jess Wilson will do a deal with One Nation," he said.
The opposition leader has explicitly ruled out forming a formal coalition with One Nation, but has refused to be drawn on whether she would rely on its MPs if they are needed to form government.
"There is no deal. There will be no deal," she said.
"The only way to change the direction of Victoria is with the majority Liberals and Nationals government."
Ms Wilson delivered the comments at the site of a damaged stretch of road in Melbourne's city centre, where she revived her pledge to fix one million potholes across the state for a cost of $5 billion over four years.
"Every time it rains in Victoria, we shouldn't see more potholes opening up on our roads because this Labor government has failed to maintain our roads for 12 years," she said.
Labor has committed to plugging more than 500,000 potholes by mid-2027 after fixing 100,000 between July and October.
Meanwhile, Mr Carroll unveiled a pledge to hire 500 additional graduate nurses and midwives.
The $132 million investment over four years would subsidise 300 graduate roles in health services across the state.
The coalition has made a series of healthcare pledges of its own, including incentives for regional workers.