Orlandi disappeared while returning home from a flute lesson in Rome on 22 June 1983.
The girl's disappearance sparked an intense media frenzy in Italy that has resulted in the case being called “Italy's most famous unsolved mystery”, and it inspired a hit Netflix four-part documentary called Vatican Girl: The Disappearance of Emanule Orlandi last year.
Emanuela was the fourth of five children of Ercole and Maria Orlandi.
Her father was a Vatican employee and the family lived inside Vatican City.
Orlandi's brother Pietro recently caused controversy by linking the case to an alleged Church paedophile ring and reporting rumours that late Saint Pope John Paul II used to leave the Vatican looking for girls with Polish cardinals.
The claim was rubbished by Vatican Secretary of State Cardinal Pietro Parolin who however said a Vatican probe into the Orlandi case would nonetheless continue.
Pietro Orlandi's lawyer, Laura Sgrò, said in a statement to ANSA after Monday's news of the fresh Rome probe: “Our wish is that there will be loyal cooperation” between the Rome Public Prosecutor's Office and the Vatican “in the search for the truth”.
“It is good news, it is what we have been asking for years to have the truth about Emanuela”.
Sgrò said she learned the news of the fresh impetus in the probe from the media and that there is still no involvement of the Orlandi family in this new phase of the investigation into the case.
Pope Francis called Pietro Orlandi's recent suggestions about the late Polish pope, who died in 2005 and became a saint in 2014, unfounded and offensive.
Speculation on Orlandi's disappearance, and that of another 15-year-old girl in the same summer of 1983, has been rife over the years.
In late November 2018 Rome prosecutors said bones found in an annex to the Vatican's nunciature to Italy did not belong to Emanuela Orlandi or the other girl, Mirella Gregori.
The Orlandi case has spawned several theories over the years, including that she was murdered to gain traction to have pope John Paul II's Turkish shooter Mehmet Ali Agca freed, or that organised crime was involved.
Ali Agca was questioned in the case.
In 2016 investigations into the case were shelved.
Six people including a priest were implicated in the investigations on suspicion of complicity in abduction and murder.
All but one had links with the Banda della Magliana, a now-defunct crime gang based in Rome.
In September 2018 the Vatican described as “false and ridiculous” reports that the Vatican had spent large amounts of money on the case.
ANSA