Tests have confirmed cases of the squirrel pox virus at three sites on an estate in the south of Scotland.
The disease, which is fatal for red squirrels but can be carried by greys without harming them, has been found on the Queensberry Estate near Dumfries.
Routine monitoring of grey squirrels confirmed the infection.
Estate manager Mark Coombs described the news as "absolutely devastating" and said the area had a "robust policy" of trying to deal with the disease.
Analysis by the Moredun Research Institute in Edinburgh found squirrel pox infected grey squirrels at three sites on the estate.
Grey squirrels are routinely sent for testing as part of a control and monitoring programme.
"We believe that the grey squirrel moved into the area along the north coast of the Solway from the Lockerbie area"
"This is worrying news," said Miles Barne, chairman of the European Squirrel Initiative (ESI).
"It is vital that squirrel control efforts are focused in this area. Squirrel pox infected greys can speed up the decline of red squirrels by up to 20 times."
The first reported case in Scotland of a red squirrel infected with squirrel pox was just over a year ago at Lockerbie.
Two grey squirrel control officers were appointed to contain the spread of infected greys.
The Queensberry Estate is over 20 miles from Lockerbie and the discovery of infected greys is seen as a blow to red squirrel conservation.
Drumlanrig Castle, 18 miles north of Dumfries and on the Queensberry Estate, has long been seen as a red squirrel stronghold.
"This news is absolutely devastating. Our red population has been safe so far," Mr Coombs said.
"Our estate gamekeepers have been controlling the grey squirrels with a robust policy of eradication to try to save this iconic native species from extinction in the wild."
Major investment
The population of greys on the estate has been growing and a recent batch of blood samples confirmed they were carrying the disease.
"We believe that the grey squirrel moved into the area along the north coast of the Solway from the Lockerbie area," said Mr Coombs.
He said it would now take a major investment from the public and private sector to tackle the problem.
The south of Scotland is one of a small number of red squirrel strongholds in the UK.
Protection officers have been appointed to try to stop the spread of the pox virus into the area.
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