England’s Mason Mount and Ben Chilwell are out of Tuesday’s game with the Czech Republic as they self-isolate because of a risk of Covid-19 exposure.
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The pair were seen interacting with Chelsea team-mate Billy Gilmour after England’s draw with Scotland on Friday – since which the midfielder has tested positive for coronavirus.
They will isolate until 28 June and could miss England’s last-16 match.
Both have tested negative for Covid-19.
England are guaranteed a place in the last 16 after Monday’s results, but will go through as Group D winners with victory over the Czech Republic.
The pair’s period of isolation will impact preparations for the knockout stage.
If England finish second in their group, their last-16 game takes place on Monday, 28 June, but if they win the group they play a day later and so the pair would be eligible.
Finishing third would see England play on either 27 or 29 June dependant on the results in other groups.
Midfielder Mount has so far played every minute of England’s European Championship campaign, while left-back Chilwell is yet to feature.
The Football Association said they will “train individually in private areas” at England’s St George’s Park training base.
After the pair initially isolated as a precaution on Monday, England manager Gareth Southgate said having Mount and Chilwell quarantined for 10 days would be the “the worst-case scenario”.
The entire national team squad had lateral flow tests on Monday and all returned negative results.
Public Health England (PHE) worked with the Football Association to confirm the duo as close contacts of Gilmour, which prompted the 10-day period of isolation.
Among Gilmour’s Scotland team-mates, however, there were no close contacts identified within the squad, with 25 players available to face Croatia on Tuesday.
PHE guidance states someone could be considered a close contact and might have to self-isolate if they had a face-to-face conversation within one metre, or had been within two metres of someone for more than 15 minutes who tested positive for Covid-19.
“We will continue to follow all Covid-19 protocols and the Uefa testing regime, while remaining in close contact with PHE,” the FA said in a statement.