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Since the players returned to limited group training on Tuesday, eight positive tests for the virus have been confirmed from England's top-flight clubs.
"The Premier League can today confirm that on Tuesday 19 May, Thursday 21 May and Friday 22 May, 996 players and club staff were tested for Covid-19. Of these, two have tested positive from two clubs," the statement said.
"The Premier League is providing this aggregated information for the purposes of competition integrity and transparency.
"Players or club staff who have tested positive will now self-isolate for a period of seven days."
The Premier League said it would not be providing details of the identities of the two new positive tests.
A total of 748 tests of players and club staff made up the initial round of testing on May 17-18. Watford's Adrian Mariappa and two members of the club's non-playing staff and Burnley assistant manager Ian Woan were among six positive tests.
Two other Watford players are also self-isolating after coming into close contact with people who tested positive, according to manager Nigel Pearson.
No matches have been played in the Premier League since March, but the government has given the green light for elite sport to resume from June 1, including the top-flight football.
The Premier League's "Project Restart" hopes to resume action in June, although no date has been fixed.
Another round of tests are scheduled to take place on Monday and Tuesday. All 20 clubs will hold a meeting on Wednesday to decide whether they can move to phase two of the restart and allow full-contact training.
Chief executive Richard Masters said on Friday the Premier League is "as confident as we can be" about a resumption of the top flight and that the remaining 92 games would be completed.
"We wouldn't have taken the first step to get back to training if we weren't convinced we had created a very safe environment for our players," he said.
Britain's death toll from the coronavirus pandemic is the highest in Europe with an official figure of 36,675 as of Saturday.
Germany's Bundesliga returned to action last week with matches in empty stadiums and strict safety protocols.
Spain's government on Saturday gave the go-ahead for La Liga to resume from June 8.
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