Deputy Prime Minister Winston Peters – speaking on behalf of Ardern in the House today – said the target was still in place and was "easily achievable".
In fact, he told media that Government would "probably build a whole lot more" than the 100,000 target.
Despite this, speaking to the Herald, Collins said it was time for Twyford to stand aside.
"Phil Twyford needs to offer his resignation to the Prime Minister and she needs to accept it."
She said he had no credibility left when it comes to KiwiBuild and the housing and urban development areas of his portfolio.
"KiwiBuild itself has been shown to be a total dog of a policy and is nothing more than a house of cards."
If he stays in the job, Collins said that would start to impact Prime Minister Jacinda Ardern's credibility and become a "massive millstone" around her neck.
Twyford's resignation would give the Government the opportunity to kill the KiwiBuild policy, she said.
Yesterday in the House, Ardern refused to confirm if the 100,000 target was still on the cards.
When questioned by media after Question Time, Twyford was also unable to confirm the figures.
"We're considering all aspects of the programme; not just KiwiBuild but the entire housing programme."
After the Government scrapped it interim KiwiBuild targets in January, Ardern and Twyford were both at pains to point out that 100,000 was still the target over 10 years.
KiwiBuild timeline:
November, 2012: Labour Leader David Shearer launches the KiwiBuild policy
2014 – 2017: Labour campaigns on KiwiBuild
November 8, 2017: The speech from the throne confirms the Government would build 100,000 homes through its KiwiBuild policy
December 19, 2017: Twyford sets up interim KiwiBuild Unit to help facilitate the building of the homes
March 25, 2018: Government purchases 29ha of land at Unitec in Auckland to build KiwiBuild homes
April 28, 2018: Construction begins on the first KiwiBuild homes
May 8, 2018: Twyford launches the "Buying off the Plans" scheme
July 4, 2018: People are invited to register their expressions of interest to buy a KiwiBuild home – close to 6000 people register interest in 24 hours
September – October, 2018: Government opens ballots on a number of KiwiBuild homes across the country
October 27, 2018: First people move into KiwiBuild home
December 18, 2018: KiwiBuild home start grant announced
January 28, 2019: KiwiBuild boss Stephen Barclay resigns amid an employment investigation – sues the Government for wrongful dismissal
January 30, 2019: Government scraps KiwiBuild interim targets and launches recalibration
May 8, 2019: Twyford and Ardern could not commit to KiwiBuild's 100,000-homes target
May 9, 2019: Judith Collins calls for Twyford's resignation and the scrapping of KiwiBuild