Seven vehicles collided near mile marker 279, six miles south of the Clermont exit, about 6:30 p.m.
But the roadway re-opened about 20 minutes later, officials said.
Three motorists were hurt, including a seriously injured woman who was airlifted to Orlando Regional Medical Center.
Two other victims were taken by ambulance to South Lake Hospital, according to FHP spokeswoman Trooper Wanda Diaz.
The damaged vehicles were removed by five tow trucks.
The crash remains under investigation.
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