Objective To examine all evidence about Microsporidia infection in vertebrate/invertebrate hosts and Iranian populations distributed in different regions of the country. Methods All published articles up to December 2015, including descriptive and cross-sectional studies related to the prevalence and genotyping of Microsporidia infection in Iran, was considered in this systematic review. The meta-analysis was done using the random-effects model and Stats Direct statistical software. MEGA 5.05 software and maximum likelihood algorithm with Kimura 2-parameter model were used for phylogenetic analysis. Results Of the 1152 investigated studies, 33 eligible studies reported a prevalence of Microsporidia infection in vertebrate and invertebrate hosts. According to this systematic review, the overall prevalence rate of Microsporidia infection in immunocompromised patients in Iran was 8.18%. Furthermore, the overall prevalence rate of Microsporidia infection in immunocompromised patients with chronic diarrhoea, patients with non-diarrhoea, gastroenteritis, and patients with CD4 (<200 cells/μL) was 15.4%, 4.1%, 0.5%, and 12.9% respectively. The highest prevalence rate of human and animal Microsporidia was estimated in Kerman (29%) and Khuzestan (26.5%). The overall prevalence rate of Microsporidia infection in honeybees using the random-effects model was 40%. Furthermore, the highest prevalence rate of nosemosis was described in East Azerbaijan (48.2%). The most Microsporidia isolates from immunocompromised patients and pigeons in Iran belonged to genotypes D (n = 16; 50%) and E (n = 6; 20.6%) of Enterocytozoon bieneusi. Conclusions This study may be the first systematic review and meta-analysis that provides a broad outlook on the prevalence of microsporidiosis in Iran. It is necessary to investigate Microsporidia infection in vertebrate and invertebrate hosts and environmental resources in Iran.