Background and Aim: Blastocystis spp. is an anaerobic, single-celled intestinal parasite with a global prevalence that is found in the lower digestive tract of humans and many animals, such as numerous types of vertebrates. To date, 17 genotypes have been identified. Nine of these genotypes are common in humans. The organism is found in both healthy and compromised individuals, hence its pathogenicity is controversial. This study aimed to investigate the frequency of Blastocystis spp. genotypes in Bandar Abbas, Iran in 2022. Materials and Methods: A total of 378 stool samples were collected from hospitals and health centers of Bandar Abbas, Hormozgan province, Iran. Stool samples were examined by direct and formalin ethyl acetate (concentration) methods. Positive samples for Blastocystis spp. were isolated and DNA extraction was done. All extracted DNA specimens were analyzed by NanoDrop to ensure quantity and quality. The PCR technique was used to identify parasite genotypes and all samples were sequenced for genotype confirmation. Results: Finally 14 samples were detected as positive microscopically and 8 samples were confirmed by PCR. Identified isolated genotypes were 4 samples ST1 (50%), 3 ST3 (37.5%), and one ST2 (12.5%). All negative and positive samples were analyzed 3 times to confirm accuracy. Conclusion: Blastocystis spp. is the most common protozoan parasite of the human intestine, and despite the studies, there are many uncertainties about its pathogenicity. The current study was designed and carried out during the COVID-19 pandemic and lower positive patients may be related to severe hygiene considerations and consumption of various antibiotics amongst the population. [ABSTRACT FROM AUTHOR]