The youth team are at home on Saturday morning (11am) when Burton Albion visit St George's Park Sheffield Graves.
The young Spireites won 2-0 at Lincoln City last week and coach Lee Walshaw is hoping they can build on that result.
He said: "The lasd have worked really hard again this week and we're looking to achieve back-to-back victories for the first time this season.
"We recorded our first clean sheet of the campaign last week and we're giving the lads challenges and incentives to work to."
Chesterfield beat Burton 2-1 in the league earlier this season before losing by the same scoreline in the FA Youth Cup at the Proact.
"They were both really tight games so it promises to be another good game," said Walshaw.