Ben Chilwell and Mason Mount are out of England’s clash against the Czech Republic and must self isolate until June 28.
The pair came into close contact with Chelsea teammate and Scotland rival Billy Gilmour after England drew 0-0 with their neighbours at Wembley last Friday.
Chilwell and Mount spent a long time in the tunnel talking with the 20-year-old, who subsequently tested positive for Covid-19.
The England duo went into isolation on Monday while a decision was made with Public Health England over how to proceed.
The Three Lions confirmed the verdict on Tuesday with a statement which provided further clarity on the situation.
“The pair were confirmed overnight as close contacts of Scotland’s Billy Gilmour after his positive COVID-19 test following last Friday’s match.
“Chilwell and Mount will isolate and train individually in private areas at England’s training base St. George’s Park, with the rest of the squad returning there after tonight’s fixture against Czech Republic at Wembley.
“We will continue to follow all COVID-19 protocols and the UEFA testing regime, while remaining in close contact with PHE.
“The entire England squad and staff had lateral flow tests on Monday and all were again negative, as was the case with Sunday’s UEFA pre-match PCR tests.
“Further tests will be carried out as and when appropriate.”
England have already secured qualification to the last 16 of Euro 2020 but this latest update plunges Mount and Chilwell’s involvement into doubt.
The duo are unavailable until Monday June 28 but England may face the Netherlands or Belgium on Sunday June 27 in the unlikely event they finish third in Group D.
If England finish top or second in the group they will play their last-16 fixture on the Monday or Tuesday, but even then it remains to be seen if Gareth Southgate would throw them into the action after a spell away from team training.
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