Senator Lambie attacked Ms Hanson's bill on Monday, saying the One Nation leader's campaign against mandatory vaccinations instilled fear in people to 'make money'.
On Tuesday she reinforced her claims to Ben Fordham on 2GB breakfast radio, telling him Ms Hanson would have to 'cope with her conscience' in proposing the bill.
'It just bothers me, when you run campaigns like that and you're putting fear into people to make money... for your political campaign, it's just not on,' she said.
'I will not play Russian Roulette with people’s lives, I'm just not into that.
'If Pauline Hanson wants to do that, she can cope with that on her conscience.'
The bill was defeated in the Senate despite the support of five government senators who crossed the floor to support it.
If successful, the bill would have removed mandatory vaccination even for those who work in aged care.
'I'm on the ground so much in Tasmania and I'm so recognisable... the majority of people who've had their vaccinations come up and they want their freedoms back, they want to get on with their lives,' Ms Lambie told Fordham.
'They’re really starting to feel the heat, they just want to get back to normal life as quickly as possible and as close as possible.
'They know like I do there is no other frontline defence [besides vaccination], this is all we've got on the table so you have a choice to take it or not to take it.'
Jacqui Lambie blasted Pauline Hanson in a furious speech in the senate as the One Nation leader watched on from QLD with a wry smile (pictured)
Ms Lambie told Fordham that business owners in her constituency could not afford to shut again.
'Those deep cleans they go into, they cost thousands of dollars... the tourism industry, the hotels and motels, they are really paying the price for this.
'They cannot afford to shut again, many of them are hanging on by a thread. They just need to get back to normal business, they can't afford to have their staff off who may be sick or anything else.'
The Tasmanian senator made headlines on Monday after accusing Ms Hanson of 'using people's fear' to gain popularity ahead of next year's election.
'Politicians like Senator Hanson and Senator Roberts are using people's fear to boost their own election campaigns and they're using fear to make money... It's all power it's all about cash and it's all about seats,' she said in her impassioned speech.
'Being held accountable for your actions isn't called discrimination, it's called being a god-damn bloody adult. It's putting others before yourself and that's what this country is supposed to be about,' she said.
Senator Lambie said state premiers introduced vaccine mandates because they 'don't want to be playing Russian roulette with other people's lives' and described One Nation as the 'enemy of health workers and officials who would have to clean up after the outbreak.'
Senator Lambie's speech came after Senator Hanson moved a bill to ban Covid-19 vaccine mandates
The 50-year-old former soldier said it was important to ask questions about the vaccine but urged anti-vaxxers to consider vulnerable people when making their decisions.
'You have a right to choose, you don't have a right to put vulnerable people's lives at risk. You don't have that right. And so you shouldn't have,' she said.
'You don't have the right to go into an aged care home unvaccinated and risk starting a Covid outbreak for the elderly.
'Nobody has the right to make someone else's life less safe. That's not what freedoms mean.
'You have the freedom to make your own choices. Everyone else has the freedom to respond to your choices. And you don't get to control that no matter how much you might want to.'
Senator Lambie also backed businesses to impose vaccine mandates on their customers, saying she knows business owners who have auto-immune diseases who would be 'risking their lives or shutting down their businesses' if mandates were banned.
Senator Lambie told 2GB that reaction to her speech 'brought out the nasties' afterwards.
'Don't abuse my staff,' she said. 'That's really uncalled for, that's really un-Australian, to do that.'
Pauline Hanson (pictured) has moved a bill to ban vaccine mandates with a passionate speech in the Senate
The speech came after Senator Hanson moved a bill to ban Covid-19 vaccine mandates.
'This legislation is urgently needed to arrest and reverse the pandemic of discrimination that has been unleashed on the Australian people,' Senator Hanson said.
The Federal Government insists the Covid vaccine is optional but state and territory governments as well as private companies have enforced vaccine mandates, requiring jabs for activities like going to work, the pub or the shops.
Senator Hanson - who is unvaccinated - wanted the Federal Government to over-ride the states and ban all vaccine discrimination.
In her passionate speech introducing the bill, Senator Hanson called Mr Morrison 'weak'.
'The Prime minister is weak, he says there should be no vaccine mandates - then do something about it,' she said.
Senator Hanson also defended pro-choice protesters, saying: 'They're not idiots, they're not ratbags, they're everyday Australians. They're not extremists.'
The 67-year-old urged other senators to support her bill which she described as 'vital'.
'We don't do this lightly. We do it with sincere regret that such legislation is even necessary. But make no mistake: it is not only necessary, but absolutely vital,' she said.
'Senators here are on notice if you don't support my legislation, then you don't support Australian democracy and freedom and you don't support the right to choose.
'We are not here to wield power against the Australian people. We are here to wield the power of the Australian people. We have no right to take away their rights.'
Prime Minister Scott Morrison and Reid MP Fiona Martin visit the Tooheys brewery in Sydney on Thursday
The bill was supported by five rogue Coalition senators Matt Canavan, Gerard Rennick, Alex Antic, Sam McMahon and Concetta Fierravanti-Wells.
But it was voted down by the Government, Labor and the Greens.
Senator Canavan, who is fully vaccinated, said no-one should be denied the right to work and put food on the table for their families if they don't want the vaccine.
'This division has to end,' he said.
'I trust Australians to be the masters of their own healthcare... without the heavy hand of Government.'
Ministers on Monday morning said the Federal Government only supports vaccine mandates in certain healthcare settings and said the states were responsible for economy-wide mandates.
Kristina Keneally said Labor supports vaccine mandates
Labor Senator Kristina Keneally slammed the Government for allowing Senator Hanson's bill to be debated and accused Mr Morrison of pandering to 'extremists' to gain votes ahead of next year's election.
'Is he willing to pander to violent extremists? Is he willing to incite division and hatred and fear? Is that the path to victory? He is pandering to One Nation.'
Senator Keneally said Labor supports vaccine mandates guided by health advice to reduce Covid deaths.
Last week Mr Morrison said vaccine mandates to enter pubs and cafes should not be in place after states reach the 80 per cent vaccination threshold.
While NSW will drop vaccine passports on December 15, Queensland will introduce them to enter hospitality venues on December 17 and Victoria has vowed to keep jab requirements in place well into next year.
Mr Morrison, who is under pressure from pro-choice politicians in his party, on Thursday said the only mandates he supports are for health workers.
In a dig at Queensland Premier Annastacia Palaszczuk, he said unvaccinated people 'should be able to go to a get a cup of coffee in Brisbane'.
'Now it's time for governments to step back and for Australians to take their life back,' he said during a visit to the Tooheys brewery in Sydney on Thursday.
'We aren't in favour of mandatory vaccines imposed by the Government. Businesses can make their own choices on the law but we aren't about telling them or Australians what to do.
'Vaccines are only mandatory in cases where you have health workers working with vulnerable people.
'That's what our medical advice has always been and, as we get above 80 per cent in particular... they should be able to go to a get a cup of coffee in Brisbane regardless of whether you've had a vaccine or not.'
Queensland Deputy Premier Steven Miles hit back at at the prime minister, accusing him of backing 'dangerous fringe elements' such as the anti-government protesters in Melbourne.
Mr Miles said Mr Morrison was undermining the state's pandemic response for his own 'cynical political interests'.
'He is so desperate to claw together a coalition of anti-vaxxers for his own political benefit that he is undermining confidence in our vaccine,' the deputy premier told parliament.
The planned venue mandate has boosted vaccine uptake, Mr Miles said, and was a deserved reward for those who had done the right thing and got the jab.
'They do not deserve to be undermined by a prime minister more interested in currying favour with coffee baron donors and lunatic backbenchers than the health and the jobs of Queenslanders,' Mr Miles said.
It comes after two Liberal