In an interview on 60 Minutes Overtime, she said she doesn't see holding office at the White House in her future.
She told the show's correspondent Ann Silvo: "I've never looked outside for other people to tell me when I should be making a move. And, wouldn't I know?" she said.
"If God actually wanted me to run, wouldn't God kind of tell me? And I haven't heard that."
Winfrey first started to get the push to enter into the next presidential election in January after her acceptance speech for Cecil B. DeMille Award at the 2018 Golden Globes in January.
The speech, which included support for #Metoo movement against sexual harassment, received a standing ovation and calls for her to start a presidential campaign.
"Are you kidding me? I was just trying to give a good speech," Winfrey told Silvo.
"I was looking for a way to express what was going on in this moment in terms of gender and class and race."
"I had a lot of wealthy men calling, telling me they would run my campaign and raise $1 billion for me," she said. "I think that when you have that many people whose opinions you value coming at you, it's worthy of thinking about."
Winfrey added that it was "humbling" that people feel she could be the next person to lead the United States, but maintained that the job was "not in her spirit".
She said: "I do feel that I have a responsibility as a person who has a big voice in this country to use it to promote justice, and kindness and goodwill in the world," she said, "but it never has felt to me that it was supposed to be political and it still does not feel that to me."
This the second time Winfrey has decline encouragement to run for president. In an interview with InStyle earlier this year, she told the magazine she doesn't "have the DNA for it."
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