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Objective: To identify the biodiversity of Nocardia species isolated from the environments of Tehran hospitals via phenotypic and PCR molecular methods. Methods: A total of 236 samples were collected from the transplantation and cancer centers of five Tehran hospitals, and the presence of Nocardia was identified by using standard protocols. Conventional tests were used for preliminary identification, and PCR amplification of the 598 bp amplicon of the 16S rRNA and sequence analysis of 16S rRNA were performed for genus and species identification. Results: From the 236 soil samples, 16 samples were identified as positive based on the culture tests, while 8 samples showed positive results based on the molecular method. The phylogenetic tree showed that the most frequent genus of Nocardia was related to Nocardia cyriacigeorgica. Conclusion: The hospital environment is a potential reservoir of a diverse range of Nocardia species. Due to the strong capability of survival, these bacteria bring a threat to the health of patients. Its identification can help physicians to implement treatments and is important for healthcare centers. [ABSTRACT FROM AUTHOR]