The stone temple was built by parishioners in 1685. Previously, there was a wooden church on this site in honor of the Nativity of the Virgin Mary with an altar dedicated to Saint Nicholas the Wonderworker.

In 1649, the residents of Nikitskaya sloboda joined the Nativity of the Virgin Mary parish. They were resettled to the devastated Kaluga during the Time of Troubles by the order of Ivan Nikitich Romanov from the trading village of Spasskoye (now Kievka). At the request of the new parishioners, in memory of the church of Spasskoye village, the altar of Saint Nicholas was replaced with the altar of the Great Martyr Nikita, hence the church received the name Nikitsky. After the fire of 1754, the church was restored in 1755. In 1820, an altar in the name of Saint John Chrysostom was added to it.

The church had a massive square base with decorative half-columns at the corners, window frames, a wide cornice, and false arches. On both sides of the altar, at the side entrances, there were galleries made of colonnades, with ten columns in each. The bell tower was octagonal and four-tiered. The Nikitsky church is recognized as a monument of architecture and urban planning of local significance (decision of the Kaluga Oblast Executive Committee No. 35 dated April 4, 1991).

From the formation of the Kaluga Diocese until the completion of the construction of the Holy Trinity Cathedral from 1799 to 1819, this church housed the cathedra of the ruling Kaluga archbishops. Several ancient icons were venerated in the church, including the image of the Great Martyr Nikita (17th century).

The church was closed in the 1920s. Initially, it was used by the military department as a warehouse. In March 1929, at the suggestion of the VKP(b) faction of the city council, the upper tiers of the bell tower and the central dome were demolished, and the church itself was repurposed. From 1939, the church housed the "Pioneer" cinema. In the 1990s, commercial structures were also located here.

Since 1993, the Diocese has repeatedly appealed for the return of the church for its restoration and the resumption of services. Only in October 2004, by the decision of the City Duma of Kaluga, the church was transferred to the Kaluga Diocese.

In January 2005, the restoration work of the church was entrusted to Deacon Alexey Pelevin.

Restoration work began in April 2005. Father Alexey faced numerous tasks: dismantling internal partitions, removing debris, interior finishing of the premises, strengthening the foundation, restoring the domes of the square base, installing crosses and domes, constructing the bell tower, acquiring church utensils, and landscaping the square next to the church.

On Easter 2005, a church shop was opened.

On September 28, 2005, after more than 80 years, the first service (a moleben with an akathist) was held in the Nikitsky church, conducted by Archimandrite Nikita (Ananyev) and Priest Alexey Pelevin.

On January 18, 2006, the first Divine Liturgy took place.

On the Annunciation in 2009, the first bell ringing sounded from the bell tower of the Nikitsky church.

A spacious Orthodox library operates at the church, where both adults and children can gather for reading. Conferences, Orthodox discussions, and meetings are held in its premises. The library fund is constantly replenished with new books.

A Sunday school has been organized for the parishioners. Classes are designed for both children and adults. A spiritual and educational center operates based on the Sunday school.