Charles and England were part of the Chelsea squad that won the FA Cup in DecemberEngland's Niamh Charles and Beth England will miss Friday's World Cup qualifier in North Macedonia after testing positive for Covid.
The Chelsea players returned positive samples in pre-camp testing and have not been in contact with their England team-mates at St George's Park.
Assistant coach Arjan Veurink will also miss the game after a positive test.
All three may be able to rejoin the set-up in time for the 12 April match against Northern Ireland.
England are five points clear at the top of Group D with six wins from six matches and a goal difference of plus 53.
Victories in their next two games would guarantee their place at next year's World Cup in Australia and New Zealand.
Head coach Sarina Wiegman has opted not to call up replacements for Charles and England, with 22 players in camp.
