It was joined by Busy Bees Castle View in Dunfermline.
The listings featured over 700 nurseries, and were based on reviews from families and carers. Nurseries were rated on overall standard, facilities and outside space, learning, resources and equipment and ICT, care, and activities.
Clair Beaton, centre director at Busy Bees Glenrothes, said: “It’s heart-warming to be recognised and makes us feel that our nursery is special just for doing the job that we all love.”
Amanda Hopkins, reviews manager at daynurseries.co.uk, said: “We would like to congratulate Busy Bees Castle View and Busy Bees Glenrothes on being rated by parents as a top nursery in Scotland.
“Being chosen as one of the best nurseries in the UK by family and carers of children that attend the setting is a huge achievement.
“Our award is recognition from parents and carers that this nursery is offering their children a high standard of care that is key to their social and emotional development as well as their learning.
“We hope these awards give parents that much-needed guidance in choosing the right nursery for their child.”
Busy Bees was formed in 1983 and opened its first nursery in 1984 in Staffordshire.
It has since grown to become the largest provider of early years childcare in the nation.