The shroud bears the figure of a crucified man, complete with blood seeping out of nailed hands and feet, and believers say Christ's image was recorded on the linen fibres at the time of his resurrection.
Scientists have reproduced the shroud using materials and methods that were available in the 14th century, the Italian Committee for Checking Claims on the Paranormal said.
The group said in a statement this is further evidence the shroud is a medieval forgery. In 1988, scientists used radiocarbon dating to determine it was made in the 13th or 14th century.
But the dispute continued because experts couldn't explain how the faint brown discolouration was produced, imprinting on the cloth a negative image centuries before the invention of photography.
Many still believe that the shroud "has unexplainable characteristics that cannot be reproduced by human means," lead scientist Luigi Garlaschelli said in the statement. "The result obtained clearly indicates that this could be done with the use of inexpensive materials and with a quite simple procedure."
The research was funded by the debunking group and by an Italian organisation of atheists and agnostics, he said.
Garlaschelli, a professor of chemistry at the University of Pavia, said in an interview with La Repubblica daily that his team used a linen woven with the same technique as the shroud and artificially aged by heating it in an oven and washing it with water.
The cloth was then placed on a student, who wore a mask to reproduce the face, and rubbed with red ochre, a well known pigment at the time. The entire process took a week, Repubblica said.
The shroud is first recorded in history around 1360 in the hands of a French knight - a late appearance that is one of the reasons why some scientists are sceptical of its authenticity.
Measuring 13 feet (4 metres) long and three feet (one metre) wide, it has suffered severe damage during the centuries, including from fires.
Owned by the Vatican, it is kept locked in a special protective chamber in Turin's cathedral and is rarely shown. The last public display was in 2000, when more than one million people turned up to see it, and the next is scheduled for 2010.
The Catholic Church makes no claims about the relic's authenticity, but says it is a powerful symbol of Christ's suffering.
The shroud has been strongly debated within the scientific community. Some researchers claim that patches used in the Middle Ages to repair the cloth after a fire altered the carbon-dating results.
Another study, by the Hebrew University, concluded that pollen and plant images on the shroud showed it originated in the area around Jerusalem sometime before the eighth century.
Garlaschelli told Repubblica he didn't think his research would convince those who have faith in the shroud's authenticity.
"They won't give up," he said. "Those who believe in it will continue to believe."