The church on the ground floor, originally planned to be the monastery refectory, was dedicated to the annunciation of the Virgin Mary and was consecrated in 1725.

From its early years it became an important burial ground for members of the imperial family, their associates, and the Russian nobility.

[2] It was initially planned that the ground floor would house the monastery's refectory, but this was altered to instead create a burial space for members of the royal family and prominent dignitaries.

Peter the Great had ordered the relics of Saint Alexander Nevsky to be sent from the Church of the Nativity of the Virgin monastery in Vladimir.

[4] The exterior walls were decorated with stucco and alabaster ornaments by Ivan and Ignatius Rossi, while the interior used blue marble.

With the completion of the Holy Trinity Cathedral of the Alexander Nevsky Lavra in 1790, the shrine and relics were transferred there at its consecration on 30 August, one of the saint's feast days.

Among them were Anna Leopoldovna, mother of Tsar Ivan VI and regent during her son's brief reign; and Emperor Peter III who was deposed and killed in a palace coup that brought his wife to the throne as Empress Catherine II.

[2] Other members of the imperial family buried in the church include Catherine Ivanovna, Peter the Great's niece; Natalia Alexeievna, Paul I's first wife; and several children who died in infancy, including two daughters of Emperor Alexander I and Elizabeth Alexeievna; Olga Pavlovna, daughter of Emperor Paul I and Maria Feodorovna.

Among them were courtiers Yevdokiya Yusupova, Anastasiya Trubetskaya, and Anna Matyushkina; as well as Alexei Razumovsky, lover and rumoured husband of Empress Elizabeth; and Maria Rumyantseva, mistress of Peter the Great.

[1][2] Repairs were carried out between 1820 and 1821, at which time a stone gateway with ornate metal doors was erected over the passage from the River Monastyrka  [ru].

[2] The memorials of Napoleonic military leaders Mikhail Miloradovich and Dmitry Senyavin were placed in the church after the closure of the Dukhovskaya Church, while those of Ivan Lazarev and Gerasim Lebedev were brought from other locations as part of a general clearing of cemeteries and the concentration of funerary monuments into centralised museum necropolises.

[7][8][9][10] In other instances parts of memorials, such as plaques and reliefs from the monuments of Alexei Turchaninov and Anna Vorontsova in the Lavra's Lazarevskoe Cemetery were installed in the church.

[11][12] In the case of sculptor Mikhail Kozlovsky, his monument was transferred from the Smolensky Cemetery to the Lazarevskoe in 1931, with some of the reliefs installed in the Annunciation Church.

[13] Some repair work was carried out during the Second World War, when a military hospital opened in the grounds of the monastery during the siege of Leningrad.

On 7 April 2013, the 300th anniversary of the monastery, vice-governor of Saint Petersburg Vasily Kichedzhi [ru] announced that the Annunciation Church would be returned to the Alexander Nevsky Lavra.