Barnsley will face Tranmere in the third round of the FA Cup on Sunday without new signings Carlton Morris and Liam Kitching.
Both players joined the Tykes this week but are cup-tied and will not feature at the weekend.
Boss Valerien Ismael confirmed that forward Conor Chaplin and goalkeeper Brad Collins have returned to training following a period of isolation due to Covid-19 exposure.
Striker George Miller was also due to return but has since sustained a knee injury and will play no part.
Tranmere boss Keith Hill has said his side are “suffering” thanks to several absences caused by injury and illness.
The club’s last scheduled fixture at Colchester on January 2 was called off due to Covid-19 issues in both squads.
Defender Manny Monthe is troubled by tendonitis and has been sidelined for Rovers’ three previous matches.
Forward Stefan Payne is a long-term absentee as he continues his recovery from a groin injury.