A record 103 Premier League players and staff tested positive for Covid in the seven days up to and including 26 December, new figures show.
The previous weekly league record was 90 cases, announced a week ago.
A total of 15,186 tests were carried out between 20 and 26 December, as the Premier League has reintroduced daily testing of players and staff.
Fifteen English top-flight games have been called off so far during December because of coronavirus cases.
A Premier League statement said: "The Premier League's Covid-19 emergency measures include protocols such as wearing face coverings while indoors, observing social distancing, limiting treatment time, as well as the increased testing.
"The league is continuing to work with clubs to keep people safe by helping mitigate the risks of Covid-19 within their squads. We are also liaising closely with the government, local authorities and supporter groups, while being responsive to any future changes to national or local guidance."
Premier League and EFL clubs recently chose to fulfil festive fixtures despite ongoing Covid-19 disruption.
The Premier League's 20 clubs met last week to discuss the fixture list following a number of postponements.
No vote took place, with two rounds of fixtures remaining in place across five days from 26 December, while the EFL will play games "where it is safe to do so".
Clubs were advised if they have 13 fit players, plus a goalkeeper, they should fulfil their games.
Manchester United were set to visit Newcastle later on Monday to play their first game in more than two weeks following a Covid outbreak within the squad.
Watford's game against West Ham on Tuesday will also be their first game since a 2-1 defeat at Brentford on 10 December.
Wolves' scheduled trip to Arsenal on Tuesday has been postponed because of Covid-19 cases in the Midlands club's camp, with Leeds United's top-flight game against Aston Villa on Tuesday has also been postponed because of an outbreak at the Yorkshire club.
Several EFL fixtures on 29 and 30 December have been called off too.