Diane Rwigara, the leader of the People Salvation Movement and an outspoken critic of Rwandan President Kagame, has been barred from contesting in next month’s presidential election.
Rwanda’s Electoral Commission claimed Ms Rwigara had failed to provide the correct documentation to show she had no criminal record.
It also said she had failed to show she had enough support nationwide to stand.
“On the requirement for 600 signature endorsements, she did not provide at least 12 signatures from eight districts,” Oda Gasinzigwa, the electoral commission chief said.
Mr Kagame will now face challengers – Frank Habineza of the Democratic Green Party and independent Philippe Mpayimana – who was cleared by the electoral body.
They were the only two cleared candidates to stand against Mr Kagame in the 2017 elections.
Ms Rwigara, who was also disqualified from the 2017 elections, expressed disappointment at the commission’s decision.
“Why won’t you let me run? This is the second time you have] cheated me out of my right to campaign,” she said.
Mr Kagame is running for a fourth term, winning the last election with 99% of the votes.
The 66-year-old president has faced criticism from rights groups for not tolerating dissent but he claims his administration respects political freedom.