The accident involved one motorcycle and happened on the A77 at the Shalloch Park roundabout just before 2pm on Tuesday, May 11.
Two men, aged 40 and 19, were taken to hospital with serious injuries, where they remain in stable condition.
The road was closed for almost four hours, reopening at 5.50pm.
The Ballantrae Coastguard Rescue Team attended the scene of the accident – just north of Lendalfoot – along with the Girvan Coastguard Rescue Team.
When the team arrived one of the two casualties was already in the care of the Scottish Ambulance Service, and the other was being tended to by a passing coastguard rescue officer.
The team from Girvan set up rescue rope equipment and recovered the second casualty with help from the Scottish Fire and Rescue Service.
The individual was then transferred into the care of the Scottish Ambulance Service.
The teams proceeded to clear away the rescue equipment and helped move both of the casualties into the waiting air and road ambulances.
A police spokesperson said: “Around 1.55pm on Tuesday, May 11, we received a report of a serious road crash on the A77 at the Shalloch Park roundabout involving one motorcycle.
"A 40-year-old man was taken to Queen Elizabeth University Hospital in Glasgow and a 19-year-old man was taken to University Hospital Ayr.
"The condition of both men is described as serious but stable.
"The road was reopened at 5.50pm.”