Ms Pelosi told reporters at her weekly press conference Thursday that as a Catholic, she resents the suggestion that she hates anyone.
She fired back at the questioner: "Don't mess with me when it comes to words like that."
The reporter asked: "Do you hate the president, Madam Speaker?"
"I don't hate anybody," Ms Pelosi responded.
She went on to call Mr Trump, the subject of only the fourth presidential impeachment inquiry in history, a "coward" on gun policy, "cruel" on immigration and "in denial" about the climate crisis.
Earlier, Ms Pelosi announced that the House will draft articles of impeachment against Mr Trump over his pressure on Ukraine to investigate Democrats and political rival Joe Biden as Mr Trump was withholding aid to Ukraine.
On impeachment, she told reporters: "This is about the Constitution of the United States and the facts that lead to the president's violation of the oath of office.
"And as a Catholic I resent your using the word hate in a sentence that addresses me."
She said she prays for Mr Trump, with whom she has faced off all year on issues like the partial government shutdown.
"I don't hate anyone. I was raised in a way that is full heart full of love and always pray for the president and I still pray for the president. I pray for the president all the time."
The question about hating Mr Trump came as she was leaving her weekly press conference.
She stopped and addressed the reporter before returning to the microphone.
"Don't mess with me when it comes to words like that," she said, and walked out.
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