The team will again be without injured captain Tomas Rosicky, who is weeks away from returning to the Arsenal lineup from an Achilles injury.
Coach Michal Bilek said he could still add players to the squad before the home matches against Malta on October 12 and Bulgaria on October 16.
The Czech attack will feature 20-year-old Watford striker Matej Vydra and 23-year-old Nuremberg player Tomas Pekhart.
The team drew 0-0 away at Denmark in their opening Group B qualifier last month.
Goalkeepers: Petr Cech (Chelsea), Jaroslav Drobny (Hamburg), Jan Lastuvka (Dnetropetrovsk)
Defenders: Theodor Gebre Selassie (Werder Bremen), Roman Hubnik (Hertha Berlin), Michal Kadlec (Bayer Leverkusen), David Limbersky (Viktoria Plzen), Frantisek Rajtoral (Vitkoria Plzen), Tomas Sivok (Besiktas), Marek Suchy (Spartak Moscow)
Midfielders: Vladimir Darida (Viktoria Plzen), Josef Husbauer (Sparta Prague), Tomas Huebschmann (Shakhtar Doneck), Petr Jiracek (Hamburg), Milan Petrzela (Augsburg), Jaroslav Plasil (Girondins Bordeaux), Jan Rezek (Famagusta)
Forwards: David Lafata (Jablonec), Tomas Pekhart (Nuremberg), Matej Vydra (Watford)