Background and aims: Mucorales are fungi belonging to the category of Zygomycetes, found much in nature. Culture-based methods for
clinical samples are often negative, difficult and time-consuming and mainly identify isolates to the genus level, and sometimes only as
Mucorales. Therefore, applying fast and accurate diagnosis methods such as molecular approaches seems necessary. This study aims at
isolating Mucorales for determination of Rhizopus genus between the isolates using molecular methods.
Methods: In this descriptive observational study, a total of 500 samples were collected from air and different surfaces and inoculated
on Sabouraud Dextrose Agar supplemented with chloramphenicol. Then, the fungi belonging to Mucorales were identified and their
pure culture was provided. DNA extraction was done using extraction kit and the chloroform method. After amplification, the samples
belonging to Mucorales were identified by observing 830 bp bands. For enzymatic digestion, enzyme BmgB1 was applied for identification
of Rhizopus species by formation of 593 and 235 bp segments.
Results: One hundred pure colonies belonging to Mucorales were identified using molecular methods and after enzymatic digestion, 21
isolates were determined as Rhizopus species. The sequencing of PCR products and macroscopic and microscopic studies confirmed the
existence of R. stolonifera, R. oryzae and R. caespitosus in the samples.
Conclusion: Generally, developing a reliable method for determining Zygomycete species can be a useful tool for better understanding of
the epidemiology of mucoromycosis.