The ministry said two previous scans by Japanese and American scientists proved inconclusive, but insisted the latest data was unambiguous.
British Egyptologist Nicholas Reeves had been the chief proponent of a theory that there was a second chamber in the tomb of Tutankhamun . This he argued means that the royal burial chamber of the queen was hidden behind the tomb of Tutankhamun and that many fabulous treasures were laying there waiting to be discovered.
A picture taken on November 28, 2015 shows the golden sarcophagus of King Tutankhamun displayed in his burial chamber in in the Valley of the Kings, close to Luxor, 500 kilometres south of the Egyptian capital, Cairo. Efforts to find a secret chamber in his tomb have led nowhere.
Completing a collection of 5,200 Tutankhamun artefacts, the Egyptian Ministry of Defence has offered the Ministry of Antiquities the sixth and last chariot of the boy king.
There is huge global interest in Nefertiti, who died in the 14th century BC and is thought to be Tutankhamen's stepmother. This seemed to provide support to the theory of Reeves that there was a secret chamber and was presumably undiscovered.
Then-Minister of Antiquities Mamdouh Eldamaty said at the time, "We can say more than 90 percent that the chambers are there".
Reeves even believed that he had detected hidden doors behind the funerary painting on the walls of the Pharaoh's tomb.
They used Ground Penetrating Radar to "see through" the thick rock tombs and search for any discontinuities between the wall's thickness.
The second team found no evidence of hidden chambers. Francesco Porcelli of the Polytechnic University of Turin will present the findings later on Sunday.
However, this new research showed no such indication of any suspicious walls, nor any large empty voids lurking behind them.
"It is concluded, with a very high degree of confidence, said Dr. Porcelli, the hypothesis concerning the existence of hidden chambers or corridors adjacent to Tutankhamun's tomb is not supported by the GPR data", it said in its statement.