"If we were officially offered the choice of Kosovo or Europe, we would renounce the European path," Nikolic told Belgrade daily Vecernje novosti.
"They can keep us at a distance for another hundred years, but we will never change our position," Nikolic stated.
"If they (the European Union) confront us with conditions which didn't apply to other members, that means they don't want us," he said, calling for greater clarity from the EU.
Belgrade opposes Kosovo's independence, declared by majority Albanians in 2008 and recognised by 90 countries including the US and most European Union countries.
But the EU has linked Serbia's membership drive to the normalisation of relations with Kosovo. This month the EU went a step further, saying Belgrade would have to recognise Kosovo's sovereignty and territorial integrity.
Serbian prime minister Ivica Dacic has said that recognition of Kosovo's sovereignty and territorial integrity is out of the question for Belgrade as it would amount to formal recognition of the statelet.
Serbia became an official candidate for EU membership in March, but the 27-member bloc has come up with fresh conditions including the abolition of so-called parallel institutions run by Belgrade in the Serb-populated north.