) is the most common parasite that can lead to a disease called toxoplasmosis. In this study, serological and molecular complementary tests have been conducted to detect or diagnose this parasite. A total of 71 patients with clinical symptoms of ocular toxoplasmosis and 20 patients with other ocular infections were evaluated. Serum and buffy coat samples were collected and tested using enzyme-linked immunosorbent assay (ELISA) and nested polymerase chain reaction (nPCR) assessments. Superficial B1 gene was evaluated in PCR. The ocular toxoplasmosis patients were followed-up 2 weeks after the first sampling and 4 weeks following the first laboratory testing. The main outcome measures were the efficiency of the diagnostic procedure and positive and negative predictive values (PPV and NPV). Overall, of the samples, 69% were PCR+, IgG+, and IgM-, and 4.2% showed PCR+, IgG+, and IgM+. In the first follow-up, after 2 weeks, from the 41 referred patients, 29 (70%) showed PCR+, IgG+, and IgM-, which confirmed the results of the first sampling. In the second follow-up, 9 (47%) patients were PCR+, IgG+, and IgM-. A correlation was observed between the first referral and the follow-ups. Also, from 71 patients, diagnosed clinically as ocular toxoplasmosis, the disease was confirmed in 73.2% and 26.8% of those suffering from other ocular infections. Of the 20 control group samples, 55% showed PCR-, IgG+, and IgM-. The sensitivity, specificity, negative and positive predictive values, and negative and positive likelihoods were analyzed for IgG and IgM antibodies and for PCR using ELISA method. As the ophthalmologic signs of may be mimicked by other infections, clinical methods may be complemented by laboratory approaches for a definite diagnosis. This would assist clinicians to achieve timely diagnosis and successful therapy and to control the infection.