The result suggests Teresa Romero, 44, is no longer infected - although a second test is required before she can be declared free of Ebola.
Ms Romero tested positive for the virus on 6 October, after she treated two missionaries who had been repatriated from West Africa. The missionaries later died from the virus.
A doctor in Madrid said she may have touched her face with her gloves after treating one of the missionaries.
The Ebola outbreak has killed more than 4,500 people across West Africa.
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