The US and UK ambassadors to the UN said technical details showed only the regime could have been responsible for the 21 August attack.
Russia said claims that the rebels were responsible could not be shrugged off.
The report said sarin-filled rockets were fired. UN Secretary General Ban Ki-moon called it a "despicable crime".
The attack led the US to threaten military action against Syria.
This sparked a diplomatic dialogue between the US and Russia, which in turn led to an agreement under which Syria has agreed to make safe its chemical weapons.
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Analysis
image of Jeremy Bowen
Jeremy Bowen
BBC Middle East editor, Damascus
I have heard some grumbling about the chemical weapons deal. Regime officials have said some are unhappy to give up what they viewed as potentially their most powerful weapon, and a deterrent of which Israel especially had to be wary.
But the key to whether President Bashar al-Assad will want to implement the agreement lies in his relationship with Moscow.
Assuming that Russia wants the Syrians to give up their chemical arsenal - and since it is giving up its own, there is no reason to doubt that - then President Assad will not want to alienate his most important friend.
The Russians sell Syria weapons and ammunition. Even more importantly, Russia has been watching Syria's back at the UN Security Council. That adds up to a lot of leverage.
Civil war far from over
'Jumped to conclusions'
The UN report did not attribute blame for the attack, as that was not part of its remit.
Mr Ban was asked at a news conference whether he knew who was behind the attack.
He replied that "we may all have our own thoughts" but it was for "others to decide" what steps should be taken to bring those responsible to justice.
UK Foreign Secretary William Hague said: "From the wealth of technical detail in the report - including on the scale of the attack, the consistency of sample test results from separate laboratories, witness statements, and information on the munitions used and their trajectories - it is abundantly clear that the Syrian regime is the only party that could have been responsible."
US ambassador to the UN Samantha Power used similar language when saying: "The technical details of the UN report make clear that only the regime could have carried out this large-scale chemical weapons attack."
She also pointed out that the 122mm rockets cited in the report were of the type previously associated with government forces and that the quality of the sarin was higher than that used by Saddam Hussein in Iraq.
French Foreign Minister Laurent Fabius said: "When you look at the findings carefully, the quantities of toxic gas used, the complexity of the mixes, the nature, and the trajectory of the carriers, it leaves absolutely no doubt as to the origin of the attack."
Mr Fabius is travelling to Moscow on Tuesday for negotiations over a possible UN Security Council resolution on the agreement to place Syria's chemical arsenal under international control.
Laurent Fabius (C): "This resolution foresees what is going to happen if the Syrians do not respect their commitments"
Russia has steadfastly refused to accept its ally Syria was to blame for the attack.
Moscow's UN envoy Vitaly Churkin said after the report was published: "I think some colleagues jumped to their conclusions when they were saying that the report definitively proves that it was the government forces that used chemical weapons."
He added: "Allegations that in fact it was the opposition who used chemical weapons cannot be simply shrugged off."
Mr Churkin questioned why no rebel fighters were among the casualties if it were a government attack.
Syrian President Bashar al-Assad has denied responsibility and blamed rebels.
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Syria's chemical weapons
CIA believes Syria's chemical weapons can be "delivered by aircraft, ballistic missile, and artillery rockets"
Syria believed to possess mustard gas and sarin, and also tried to develop more toxic nerve agents such as VX gas
Syria has not signed the Chemical Weapons Convention (CWC) or ratified the Biological and Toxin Weapons Convention (BTWC)
Sources: CSIS, RUSI
Western military options
Media review: Mixed response
UN resolution
Earlier on Monday, Mr Ban briefed the Security Council on the report.
He said: "The mission has concluded that chemical weapons were used on a relatively large scale in the Ghouta area of Damascus [on 21 August]. The attack resulted in numerous casualties, particularly among civilians."
The UN investigators examined blood, hair, urine and rocket samples.
Mr Ban said 85% of the blood samples tested positive for sarin.
He said: "On the basis of its analysis, the mission concluded that it - and I quote - 'collected clear and convincing evidence that surface-to-surface rockets containing the nerve agent sarin were used in the Ein Tarma, Moadamiyah and Zalmalka in the Ghouta area of Damascus.'"
He said the mission was unable to verify the number of casualties, but referred to the "terrible loss of life on 21 August".
Mr Ban added: "This is the most significant confirmed use of chemical weapons against civilians since Saddam Hussein used them in Halabja in 1988."
The UN powers will now try to hammer out a Security Council resolution on the deal brokered at the weekend by Russia and the US under which Syria will disclose its chemical weapons within a week and eliminate them by mid-2014.
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