The Mexican patient was also released from a hospital in Hong Kong on Friday after a "thorough clinical assessment", officials said.
The hotel guests were faced with dozens of reporters, photographers and camera crews as they left the building.
"I'm out, I didn't like it, not at all, but I am very relieved now," Kevin Ireland, a businessman from India, said.
"I think most people felt that it was too much but the government must what the government must do."
Critics have suggested the mass quarantining of hotel guests was a political move, but local authorities have repeatedly defended it in light of the Sars (severe acute respiratory syndrome) outbreak in 2003.
The disease killed 300 people in Hong Kong and 800 across the world after one carrier spread the disease in a city hotel.
"We have been successful in our effort to prevent the spread of the virus, but this doesn't mean that we can relax our guard," York said.
The 25-year-old Mexican who was diagnosed with H1N1 flu was Asia's first case of the virus, which is confirmed to have killed at least 42 people in his home country.
Hong Kong authorities released its first batch of 35 people on Thursday, most of whom had come into contact with the infected man on a flight he took from Shanghai to Hong Kong.
The World Health Organisation (WHO) said on Friday that 2,384 cases of H1N1 had been confirmed in 24 countries.
As well as the deaths in Mexico, the virus has also claimed the lives of two people in the United States.