The Azzurri have one foot in next summer's World Cup and can confirm their place with a result against Michel Bilek's side.
Cesare Prandelli's side put itself on the verge of qualification with a 1-0 win over Bulgaria in Palermo on Friday and could become the first European nation to book a ticket to Brazil on Tuesday.
Alberto Gilardino's goal moved the team seven points clear at the top of Group B and Italy will fancy its chances of adding to its 13 goals this qualification campaign against a Czech side that shipped two goals against Armenia.
Those two strikes saw Michel Bilek's side fall to a costly 2-1 defeat that could ruin its chances of reaching the World Cup, as the team missed a glorious opportunity to move into second place.
Italy has injury problems ahead of the game, with Luca Antonelli (ankle) and Ignazio Abate (groin) both ruled out of the match, while Lorenzo Insigne is a doubt.
Manuel Pasqual and Lorenzo De Silvestri have been called up to replace the injured pair, while Pablo Osvaldo and Riccardo Montolivo are also available after serving their bans.