Sale's final-round English Premiership match against Worcester on Wednesday has been called off after a new clutch of Sale players tested positive for coronavirus.
The match was originally due to be played on Sunday but was postponed after 16 players and three backroom staff from the title-chasing Sharks tested positive last week.
The Premiership said in a statement that an additional six players and two staff from the club had tested positive for Covid-19 after a new round of testing on Tuesday.
A PRL (Premiership Rugby Limited) panel met on Wednesday and in light of the new positive tests, which takes the club's total to 27, decided to cancel the game.
Sale's quest to reach the play-offs is now over after they were forced to forfeit the game, with a 20-0 win awarded to Worcester.
Saturday's semi-finals will see Wasps host Bristol while Bath travel to Exeter.
Premiership Rugby chief executive Darren Childs said: "We know everyone at Sale Sharks will be devastated, and we share their disappointment that their season ends this way.
"But we can't jeopardise the health of any players, staff or management at any of our clubs."
The club said in a statement: "This is devastating for Sale Sharks, the players and staff, with Covid-19 denying the club the chance to challenge for the Premiership title at the final hurdle."
But they made another strong rebuttal of reports suggesting the outbreak was the result of a drinking session as the squad celebrated victory in the Premiership Rugby Cup final last month.
"Reports over the weekend suggesting our players acted irresponsibly by partying and socialising throughout Manchester after our Premiership Rugby Cup victory are completely inaccurate and untrue," the statement said.
"Health and safety has always been our top priority throughout the return of professional rugby in England and we have an important role to play in supporting the Premiership Rugby players that have tested positive for this terrible virus."
The club have been the focus of an investigation by the Rugby Football Union, which revealed on Wednesday that it had "concerns" over the situation at Sale.
"The RFU is undertaking a full and thorough investigation and is working closely with Public Health England," a statement said.
"A summary of findings will be published in due course and we will consider if there are any learnings for all clubs.
"The RFU has concerns about how this situation has developed and will be making recommendations to PRL and the PGB (Professional Game Board) to avoid future uncertainty for clubs and players as well as protecting the reputation of the game."
The cancellation of what would have been the last match in the league, left the Sharks fifth in the final table, three points behind fourth-placed Bath while Worcester finished ninth.