His grandson Brady Lukas revealed the news on Wednesday on his Twitter account, saying: "Wayne recently tested positive for COVID-19 and has since been isolating at home and following CDC and health care provider recommendations.
"Fortunately, he had not been at work for several days prior to the first symptom, so no one else at the barn is impacted."
He added: "Wayne is on the road to recovery and looks forward to getting back to the track."
D Wayne Lukas, who turns 85 on September 2, is one of the greatest trainers in US racing history with 14 Triple Crown races – among them four Kentucky Derbys – and 20 Breeders' Cup events to his name.
A four-time Eclipse Award-winning trainer, Lukas has more than 4,800 career victories in North America according to Equibase. He also trained 739 quarterhorse winners before turning to thoroughbreds.
The Hall of Famer did not travel his horses to Saratoga this summer for the first time in decades, preferring to stay at his Churchill Downs base.
Meanwhile, Virginia track Colonial Downs cancelled its scheduled cards on Tuesday and Wednesday after jockey Trevor McCarthy tested positive for coronavirus.
The rider was taken off his mounts at the weekend when he experienced what were described as "flu-like symptoms" but was unable to get a test until Tuesday.