The Blairhall Primary youngsters were being taken to a netball match when the vehicle collided with a lorry on the Oakley to Saline road near Dunfermline.
One teacher, who was driving the bus, had to be cut free and was taken to Queen Margaret Hospital in Dunfermline.
The crash happened at 0940 GMT and Fife Council has said that all the children had been wearing seatbelts.
The bus was carrying 11 primary seven pupils and two members of staff.
Two children were treated for only minor injuries.
One of the adults has been discharged while the second remains in a stable condition in hospital.
Head of education services, Ken Greer, said: "The parents of the children involved have all been notified.
"Education service psychologists are at the school now to help the children and staff with any possible trauma."
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