Miliband said: "My position is no, we don't want an in/out referendum."
"My postion is precisely the same as his position when we voted together in October 2011 against an in/out referendum. My position hasn't changed, it's his position that has changed."
The Labour leader's categorical statement that he does not want an in/out referendum appears to give him little wriggle room to change his mind at a later date - meaning any U-turn on the issue would be highly embarrassing.
Tory MPs cheered wildly as Cameron entered the Commons chamber, giving the prime minister the best reception in a long time.
Cameron, who had one of his easiest PMQs for weeks, managed to get Miliband to come down against a referendum by quizzing him on Labour's position.
"We want a renegotiation and then a referendum. What does he want? Or doesn't he know?" he said.
However due to the noise in the Commons it appeared the prime minister failed to notice Miliband's unequivocal reply.
Instead Cameron repeated the attack that the Labour leader had an inability to say whether he supports a referendum or not - despite the fact he just had.
The prime minister has said he will try to renegotiate Britain's relationship with the EU and then campaign fo a 'Yes' vote in a subsequent referendum.
However he has so far avoided saying how he would vote if he was unsuccessful in winning back powers from Brussels.
Miliband told MPs: "The reason the people behind him [Tory MPs] are cheering is not because they want to vote 'Yes' in an in/out referendum, it's because they want to vote 'No'."
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