Obama, unsmiling and choosing his words carefully, said: "At a certain point if what somebody says contradicts what you believe so fundamentally and then he questions whether or not you believe it - in front of the National Press Club - then that's enough.
"That's a show of disrespect to me. It is also, I think, an insult to what we've been trying to do in this campaign," he added. "Whatever relationship I had with Reverend Wright has changed as a consequence of this."
Wright, asked on Monday about a speech in which he asserted the September 11 attacks were retaliation for U.S. foreign policy, said: "You cannot do terrorism on other people and not expect it to come back to you."
Asked about another sermon in which he suggested the U.S. government created the AIDS virus to kill black people, Wright said: "Based on what has happened to Africans in this country, I believe our government is capable of doing anything."
Wright, who presided over Obama's wedding and baptized his children, is now semi-retired from Chicago's Trinity United Church of Christ.
"The person that I saw yesterday was not the person that I met 20 years ago," Obama said. "His comments were not only divisive and destructive but I believe that they end up giving comfort to those who prey on hate."
Obama said he liked the church's new pastor and was still a member.
Obama has denounced Wright before and is now adapting his campaign to reintroduce himself to working-class voters who handed wins to Clinton in Ohio and Pennsylvania.
Matthew Wilson, a political scientist at Southern Methodist University in Dallas said the strong comments came late.
"A denunciation at this stage appears to be politically motivated because Rev. Wright didn't say anything yesterday that he hasn't been saying for 30 years," Wilson said.
