Stool samples from 150 immunocompromised patients (including 120 Gastrointestinal [GI] symptomatic and 30 GI asymptomatic) suspected to have Cyclospora cayetanensis infection were examined to evaluate the validity of Miláček-Vitovec staining method in comparison with kinyoun stain for the diagnosis of cyclosporiasis. All results were confirmed with nested PCR as a gold standard. Validation parameters included sensitivity, specificity, accuracy index, positive predictive value and negative predictive value. In addition, infection intensity was compared among all positive samples. DNAs of Cyclospora oocysts were detected among 30/120 (25%) of GI symptomatic patients versus 1.7 % of GI asymptomatic (5/30). Miláček-Vitovec stain showed more validity than kinyoun with a sensitivity reached 80% versus 73.4% (28 versus 25 from 35 cases) while the specificities of showed 90.4% (104/115) versus 81.7% (94/115), respectively. Mild infection intensity was detected among 10 patients only three of them (30%) were detected by Miláček-Vitovec stain while only one (10%) was detected by Kinyoun, Moderate infection were identified among 10 patients all of them were diagnosed by Miláček-Vitovec stain and 9 (90%) was detected by Kinyoun stain. Heavy infection was detected among 15 patients all diagnosed with Miláček-Vitovec stain while 14 were detected with kinyoun.