Boris Johnson says the EU must "urgently clarify its intentions" over its decision to temporarily override part of the Brexit deal amid a growing row about Covid-19 vaccine supplies.
The EU triggered a clause to introduce export controls on vaccines to Northern Ireland.
Under the Brexit deal, all products should be exported from the EU to NI without checks.
The EU said the move was "justified" to avert issues caused by lack of supply.
On Friday, the bloc invoked Article 16 of the Northern Ireland Protocol in a bid to prevent the region becoming a backdoor for EU vaccines to be sent to the wider UK.
The BBC's Europe editor Katya Adler said EU sources have told her that the European Commission was going to backtrack on its decision and would not suspend part of the Northern Ireland Protocol.
The source told our correspondent that "suggesting to do that was misjudgement and an error".
In the new regulation, the European Commission stated: "This is justified as a safeguard measure pursuant to Article 16 of that Protocol in order to avert serious societal difficulties due to a lack of supply threatening to disturb the orderly implementation of the vaccination campaigns in the Member States."
The EU's move should not directly disadvantage Northern Ireland, which gets its vaccine supplies through the UK procurement system.
The EU's actions have been criticised by a string of politicians, with Northern Ireland's First Minister Arlene Forster describing it as "an incredible act of hostility" that places a "hard border" between Northern Ireland and the Republic of Ireland.
Cabinet minister Michael Gove told his opposite number on the EU-UK Joint Committee, Maros Sefcovic earlier that the UK was concerned by the "lack of notification from the EU about its actions in relation to the NI protocol" and warned that Britain "would now be carefully considering next steps".
In a phone call with the Irish Taoiseach, Micheál Martin on Friday evening, Mr Johnson "set out his concerns" about the move and "what these actions may mean for the two communities in Northern Ireland", according to a No 10 spokesperson.
The PM is also said to have "stressed the UK's enduring commitment" to the Good Friday agreement and called on the EU to "urgently clarify its intentions and what steps it plans to take to ensure its own commitments with regards to Northern Ireland are fully honoured".
In a statement released on Friday evening, a No 10 spokesman said Mr Johnson had spoken to EU Commission President Ursula von der Leyen, and expressed his "grave concerns" about the "potential impact" of the EU's actions on vaccine exports.
It is the latest development in a deepening dispute over the vaccine producer AstraZeneca's delivery commitments to the EU.
The bloc agreed to buy up to 400m doses of the AstraZeneca vaccine last year, and on Friday the EU's drugs regulator approved the vaccine's use for all adults.
But the firm said that due to problems at one of its EU factories, supplies would be reduced by about 60% in the first quarter of 2021.
In an interview with the Times, Michel Barnier, who was the EU's chief Brexit negotiator, said he was calling for "cooperation" between the EU and the UK over the vaccine supplies across Europe.
He said the world was facing an "extraordinarily serious crisis" which he argued must be faced with "responsibility" rather than the "spirit oneupmanship or unhealthy competition".
He added: "I recommend preserving the spirit of co-operation between us."
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2021-01-29 22:19:00Z
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