Four people in Spain, including the nurse who tested for Ebola, have been hospitalised and are being monitored over suspicion of potential contagion.
Officials for Madrid's health system told a press conference those hospitalised included the nurse's husband, a traveller from one affected country and another health worker.
They also said the nurse, who did not leave Madrid during her holidays, was currently being treated with drip using antibodies from previous infected patients.
The European Union has asked Spain to explain how the nurse contracted the deadly disease, the first known case of transmission outside Africa.
The European Commission "sent a letter Monday to the Spanish health minister to obtain some clarification" of how this had happened, despite all the precautions taken, a spokesman said.
Yesterday, Spanish Health Minister Ana Mato said an emergency protocol had been put in place and authorities were working to establish the source of the contagion, in the first case of Ebola being contracted outside of west Africa.
"She is a health professional who took care of the infected with the disease who were repatriated and cared for at Carlos III" hospital, the director of Spain's public health department, Mercedes Vinuesa, told a news conference.
The nurse began to feel sick on 30 September. She is said to be in a stable condition.
She had treated Fr Manuel Garcia Viejo, 75, at the Madrid hospital when he was repatriated from Sierra Leone with the disease.
Fr Garcia Viejo died days later, the second Spanish priest to die after being repatriated from Africa with the disease.
The nurse went on holidays on 25 September after the priest's death.
With concerns growing around the world of the Ebola pandemic spreading beyond West Africa, Spanish officials sought to reassure the public that they were tackling the threat.
Twenty-two people who came into contact with the nurse are being monitored, said Rafael Perez-Santamaria, head of the Carlos III Hospital where the infected nurse treated the two Spanish missionaries.
They have not been isolated but they are having their temperature taken twice a day to check for signs of infection, he added.
Officials said they were still investigating how the nurse was infected.
"This has taken us by surprise," said Mr Perez-Santamaria. "We are revising our protocols, improving them."
Norwegian doctor treated for Ebola
Norway has repatriated a doctor who contracted Ebola in Sierra Leone while working for Doctors Without Borders (MSF), and whom the medical charity described as "in good spirits".
The patient - a 30-year-old woman, according to medical sources was transported in a French air ambulance and then taken to the Oslo University Hospital, where she will remain in total isolation and receive experimental treatment.
Norway is currently trying to obtain the last available dose of the prototype ZMapp drug, senior drug official Steinar Madsen told specialised website Dagens Medisin.
Other experimental treatments, like Avigan and TKM-Ebola, have also been ordered.
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