Visit Czech Republic confirmed the changes took effect on Saturday (9 April). "There are no longer any coronavirus restrictions for entry into the Czech Republic," it said.
The changes apply to travellers arriving both from EU and non-EU countries.
The country’s interior ministry added: "As of 9 April 2022, the protective measures regarding the conditions of entry into the Czech Republic in relation to the epidemic of Covid-19 have been suspended.
"Entry into the Czech Republic is no longer subject to any special epidemiological conditions to prevent the spread of the disease. The entry-ban for foreigners from third-countries and the obligation to prove infection-free status have been lifted."
Katarina Hobbs, CzechTourism’s UK and Ireland director, said: “On behalf of CzechTourism UK and Ireland, we are very happy to confirm that all Covid-19 entry restrictions have been abolished as of 9 April.
"Travellers from all over the world are welcome to travel comfortably again without stressful procedures and can experience the wonderful spring season in the Czech Republic this year."
The Foreign Office has also updated its advice to reflect the change.
"From 9 April 2022, the Czech Republic is lifting all Covid-19 entry requirements for people arriving from other countries," said the FCDO. "Travellers will no longer be required to fill in arrival forms or produce proof of vaccination."