‘Re-analysis of Carter's samples from Beijing 2008 resulted in a positive test for the prohibited substance methylhexaneamine,’ the International Olympic Committee said in a statement Wednesday.
‘The Jamaican team is disqualified from the men's 4x100m relay event. The corresponding medals, medallist pins and diplomas are withdrawn and shall be returned.’ Carter's positive test disqualifies the Jamaican team from the race and leaves Bolt - who celebrated the unique triple triple of 100m, 200m and 4x100m victories at the Rio Games last year - with eight Olympic golds instead of nine.
As a result, Finnish distance runner Paavo Nurmi and American sprinter Carl Lewis remain the most successful Olympians in athletics with nine golds each.
Bolt said in August after completing the Rio treble he would have to live with it should he lose the 2008 relay gold.
‘I don't think my legacy would be tarnished. I have proved it over and over again that I have done it clean, so it's no stress,’ he said.
‘It's not what I want, but it's life. What are you going to do? It's not something I have control over.’ The IOC is reanalysing more than 1,000 doping samples from the 2008 and 2012 Games using improved analytical methods with several athletes already disqualified retrospectively.
On Wednesday the IOC also said the Russian Tatiana Lebedava, who won silver in the Beijing long jump, had been disqualified due to a positive test for dehydrochlormethyltestosterone (turinabol).
The 31-year-old Carter was also a team-mate of Bolt in winning the 4x100m relay at the 2012 London Olympics, and took relay golds at the world championships in 2011, 2013 and 2015.
Those victories remain unchanged by Wednesday's ruling although Carter's doping sample from 2012 is sure to be retested by the IOC. The Athletics governing body IAAF could also impose sanctions.
Trinidad and Tobago are in line for the 2008 relay gold with Japan moving into silver and Brazil into the bronze medal position.
Losing the gold means Bolt is no longer tied with the great American sprinter Carl Lewis on nine at the Olympics.
But Bolt will attempt to add to his current tally of 11 titles at the world championships in London this August, in what is the highlight of what is expected to be his final season in athletics.