China took the drastic decision to cut off the city of 11 million people on January 23 after it became clear that the virus was highly contagious and potentially deadly.
The decision to lift the ban - albeit with travelers required to meet certain conditions - came despite fears of a second wave of infection if restrictions were eased too soon, Al Jazeera reported.
Within hours of the lifting of the ban, roughly 65,000 people had left the city by train and plane alone, according to the The Associated Press news agency news agency, quoting local media reports.
The first train to carry departing passengers out of the city left at 00:50 am on Wednesday (1650 GMT on Tuesday) and outbound highways were opened to traffic about the same time.
About 80 percent of the official number of fatalities in China, or about 2,571, were recorded in Wuhan.
At its peak, the lockdown saw people forcibly confined to their homes and the transport and industrial hub resembled a ghost town with streets deserted except for police patrols and emergency vehicles.
But such restrictions have eased in recent days as new infections dropped to a trickle.
Still, authorities are walking a fine line between allowing greater freedom of movement and economic activity while also guarding against a second wave of infection, with particular concern around imported cases and people who show no symptoms but can still pass on the virus.
Wuhan residents were still being urged not to leave their neighborhoods, the city and even the province unless necessary, health officials said on Tuesday.
China on Wednesday reported 62 new virus cases, 59 of them brought from outside the country and two additional deaths.
The country where the virus first emerged now has recorded 3,333 deaths and 81,802 total cases, with 1,190 people remaining in treatment, 189 of them in serious condition.
Another 83 suspected cases and 1,095 people who have tested positive but show no symptoms remain under isolation and monitoring.