Previously, officials said they were "90 per cent sure" of a hidden room behind the wall of the boy king's famous 3,000-year-old tomb, reports the BBC.
One theory suggested it could have been the tomb of Queen Nefertiti - who some think was Tutankhamun's mother.
New research, however, has concluded the chamber simply is not there.
The search for the hidden tomb began when English archaeologist Nicholas Reeves, examining detailed scans of the chamber, discovered what looked like faint traces, or "ghosts", of doors beneath the plaster.
In his 2015 paper "The Burial of Nefertiti", Reeves argued that the relatively small tomb had originally been designed for Queen Nefertiti and her remains could possibly lie further within the tomb.
Nefertiti's remains have never been discovered, but she has been the object of much speculation. A 3,000-year-old sculpture of the queen, immaculately preserved, has made her one of the most recognisable women of ancient Egypt.
It is also thought she may have ruled Egypt as pharaoh herself between the death of her husband and the ascension of Tutankhamun.
After Reeves' paper, a series of radar scans seemed to support his theory.
A second scan also seemed to support the theory, which would have been the most significant discovery of Egyptian antiquities in decades.
However, Italian specialists from the University of Turin used new penetrating radar scans to reach their conclusion, saying they were confident of the results, the BBC reported.
"It is maybe a little bit disappointing that there is nothing behind the walls of Tutankhamun's tomb," said Francesco Porcelli, head of the research team.
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